IL-12 heterodimer FC-fusion protein

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The novel IL-12-Fc fusion protein with modified subunits addresses the half-life and toxicity issues of recombinant IL-12, enhancing stability and efficacy for cancer therapy.

JP2026082822APending Publication Date: 2026-05-19XENCOR INC

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Patent Information

Authority / Receiving Office: JP · JP
Patent Type: Applications
Current Assignee / Owner: XENCOR INC
Filing Date: 2026-01-05
Publication Date: 2026-05-19

Application Information

Patent Timeline

05 Jan 2026

Application

19 May 2026

Publication

JP2026082822A

IPC: C07K14/54; A61K38/20; A61P37/02; C07K19/00; C07K16/00; C12N15/62; C12N15/24; C12N15/13

CPC: A61K38/00; C07K14/5434; C07K2319/30; A61K38/208; C07K1/18; A61P35/00; C07K19/00; A61K47/6813

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Antibody mimetics/scaffolds Peptide/protein ingredients

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AI Technical Summary

Technical Problem

Recombinant IL-12 has a short half-life and faces toxicity issues in human clinical trials, necessitating frequent injections and hindered by the IL-12p40 subunit forming homodimers that antagonize its activity.

Method used

A novel IL-12-Fc fusion protein with modified IL-12p40 and IL-12p35 subunits, featuring specific amino acid substitutions to enhance stability and reduce toxicity, promoting heterodimerization and affinity for IL-12 receptors.

Benefits of technology

The modified IL-12-Fc fusion protein extends half-life and reduces toxicity, providing a stable and effective cytokine-based therapy for cancer treatment.

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Abstract

A composition containing the variant IL-12p40 subunit domain is provided. [Solution] The present invention provides a composition comprising a variant IL-12p40 subunit domain, wherein the variant IL-12p40 subunit domain has the amino acid substitution E59K / K99Y / C252S.

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Description

Technical Field

[0001] Cross - reference to related applications This application claims the benefit of U.S. Provisional Patent Application No. 62 / 740,813, filed Oct. 3, 2018; U.S. Provisional Patent Application No. 62 / 810,038, filed Feb. 25, 2019; and U.S. Provisional Patent Application No. 62 / 848,512, filed May 15, 2019, all of which are hereby incorporated by reference in their entirety for all purposes.

Background Art

[0002] For T cells to initiate an effective anti - tumor response, three things must occur. First, T cells must engage antigenic tumor peptides presented by MHC in the tumor environment. Second, co - stimulatory molecules must bind to the T cells. And third, T cells must be induced by cytokines such as IL - 12 and IL - 2 to produce co - stimulatory cytokines such as IFNγ that enable differentiation and proliferation. Recognition of a single tumor peptide in the absence of cytokine induction causes T cells to become anergic, thereby resulting in tolerance. Thus, a very promising approach in cancer immunotherapy is cytokine - based therapy. In fact, IL - 2 is approved for use in patients with metastatic renal cell carcinoma and malignant melanoma. However, currently, there is no approved use of recombinant IL - 12 in humans.

[0003] Recombinant IL - 12 is a promising cytokine - based therapy due to its broad effects in activating the immune system. However, IL - 12 has thus far faced hurdles in human clinical trials due to toxicity. Like other cytokines, the short half - life of IL - 12 requires frequent bolus injections.

[0004] In addition, IL-12 consists of an α-chain (p35 subunit, IL-12p35) and a β-chain (p40 subunit, IL-12p40) covalently linked to form a biologically active IL-12 heterodimer. IL-12 exerts its cellular signaling function through binding to the dimeric IL-12 receptor complex, which consists of IL-12 receptor β1 (IL-12Rβ1) and IL-12 receptor β2 (IL-12Rβ2) on T cells, and inducing IFNγ secretion. However, the IL-12p40 subunit can also exist as a homodimer, which has been reported to antagonistize IL-12 activity by competing for binding to the IL-12 receptor.

[0005] This invention addresses the short half-life of IL-12 by providing a novel IL-12-Fc fusion protein and a novel IL-12 variant with reduced efficacy. [Overview of the project]

[0006] In one embodiment, the present invention provides a heterodimer Fc fusion protein comprising: a) a first fusion protein comprising a variant IL-12p40 subunit domain and a first Fc domain, wherein the IL-12p40 subunit domain is covalently bonded to the N-terminus of the first Fc domain; and b) a second fusion protein comprising an IL-12p35 subunit domain and a second Fc domain, wherein the IL-12p35 subunit domain is covalently bonded to the N-terminus of the Fc domain, wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains. In some embodiments, the present invention provides a heterodimer Fc fusion protein comprising: a) a first fusion protein comprising a variant IL-12p40 subunit and a first Fc domain, wherein the IL-12p40 subunit is E59K, E59Q, D The present invention provides a heterodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of 18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0007] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E5 9K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K This invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0008] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL -12p40(Q42E)), vii) Sequence ID 63(IL-12p40(E45Q)), viii) Sequence ID 64(IL-12p40(Q56E)), ix) Sequence ID 65(IL-12p40(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67(IL-12p40(Q42E / E45Q)), xii) Sequence ID 68(IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70(IL-12p40(Q56E / E59Q)) xv) Sequence ID 71 (IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii )Sequence ID 79 (IL-12p40(E33Q)), xxiv)Sequence ID 80 (IL-12p40(Q229E)), xxv)Sequence ID 81 (IL-12p40(E235Q)), xxvi)Sequence ID 82 (IL-12p40(Q256N)), xxvii)Sequence ID 83 (IL-12p40(E299Q)), xxviii)Sequence ID 84 (IL-12p40(D87N)), xxix)IL-12p40(E32Q), xxx)IL-12p40(D34N), xxxxi)IL-12p40(S43E), xxxii)IL-12p40(S43 K), xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)),xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlv iii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv ) Sequence ID XXX(IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX(IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX(IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX(IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence number Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)),lxvii) Sequence ID XXX (IL-12p40(N103D / N113D, )), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX(IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX(IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX(IL-12p40(N103Q)), lxxviii) Sequence ID XXX(IL-12p40(N113Q)), lxxix) Sequence ID XXX(IL-12p40(N200Q)), lxxx) Sequence ID XXX(IL-12p40(N281Q)), lx xxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL -12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E)),xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K 264E)), ciii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cv The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of ii) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0009] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the variant IL-12p40 subunit has at least 90% identity with a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0010] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the IL-12p35 subunit is a variant IL-12p35 subunit.

[0011] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0012] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the variant IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0013] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the variant IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0014] In some embodiments, the present invention relates to the variant IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151 D)), viiii) Sequence ID 120 (IL-12p35(Q135E)), ix) Sequence ID 121 (IL-12p35(Q35D)), x) Sequence ID 122 (IL-12p35(Q146E)), xi) Sequence ID 123 (IL-12p35(N76D)), xii) Sequence ID 124 (IL-12p35(E162Q)), xiii) Sequence ID 125 (IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX (IL-12p35(D55Q)), xvi) IL-12p35(E79Q), x vii) IL-12p35(Q130E), xviii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230(IL-12p35(N151D / K158E)), xxviii) Sequence ID 231(IL-12p35(E79Q / N151D)), xxix) Sequence ID 232(IL-12p35(D55Q / N151D)), xxx) Sequence ID 233(IL-12p35(N136D / N151D)), xxxx) Sequence ID 234(IL-12p35(N21D / N151D)), xxxii) Sequence ID 235(IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX(IL-12p35(F96A)), xxxiv) Sequence ID XXX(IL-12p35(M97A)), xxxv) Sequence ID XXX(IL-12p35(L89A)), xxxvi) Sequence ID XXX(IL-12p35(L124A)), xxxvii) Sequence ID XXX(IL-12p35(M125A)), xxxviii) Sequence ID XXX(IL-12p35(L75A)), xxxiv) Sequence ID XXX(IL-12p35(I171A)), xxxv) Sequence ID XXX(IL-12p35(N71Q )), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35(N85Q / N195Q), xli) Sequence ID XXX (IL-12p35(N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35(N71D / N85D)),This invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D), xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0015] In some embodiments, the present invention provides a heterodimerized Fc fusion protein in which the modification that promotes heterodimerization of the first and second Fc domains is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q;T366S / L368A / Y407V and T366W;T366S / L368A / Y407V / Y349C and T366W / S354C.

[0016] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the variant IL-12p40 subunit domain is bound to the first Fc domain using a first domain linker and / or the IL-12p35 subunit domain is bound to the second Fc domain using a second domain linker.

[0017] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0018] In some embodiments, the present invention relates to the first and / or second Fc domain However, according to EU numbering, the present invention provides a heterodimeric Fc fusion protein having an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K.

[0019] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0020] In some embodiments, the present invention relates to the heterodimer Fc fusion protein,

[0021] a)i) Sequence ID 47 (XENP27201 Chain 1), ii) Sequence ID 85 (XenD24752), iii) Sequence ID 86 (XenD24753), iv) Sequence ID 87 (XenD24754), v) Sequence ID 88 (XenD24755), vi) Sequence ID 89 (XenD24756), vii) Sequence ID 90 (XenD24757), viiii) Sequence ID 91 (XenD24758), ix) Sequence ID 92 (XenD24759), x) Sequence ID 93 (XenD24760), xi) Sequence ID 94 (XenD24761), xii) Sequence ID 95 (XenD24762), xiii) Sequence ID 96 (XenD24763), xiv) Sequence ID 97 (XenD24764), xv) Sequence ID 98 (XenD24765), xvi) Sequence ID 99 (XenD24766), xvii) Sequence ID 100 (XenD24767), xviii) Sequence ID 101 (XenD24768), xix) Sequence ID 102 (XenD24769), xx) Sequence ID 103 (XenD24770), xxi) Sequence ID 104 (XenD24771), xxii) Sequence ID 105 (XenD24772), x xiii) Sequence ID 106 (XenD24773), xxiv) Sequence ID 107 (XenD24774), xxv) Sequence ID 108 (XenD24775), xxvi) Sequence ID 109 (XenD24776), xxvii) Sequence ID 110 (XenD24777), xxviii) Sequence ID 111 (XenD24778), xxix) Sequence ID 112 (XenD24792), xxx) Sequence ID 215 (XenD25922), xxxxi) Sequence ID 216 (XenD25923), xxxii) Sequence ID 217 (XenD25924), xx xiii) Sequence ID 218 (XenD25925), xxxiv) Sequence ID 219 (XenD25926), xxxv) Sequence ID 220 (XenD25927), xxxvi) Sequence ID 221 (XenD25928), xxxvii) Sequence ID 222 (XenD25929), xxxviii) Sequence ID 223 (XenD25930), xxxix) Sequence ID 224 (XenD25931), xl) Sequence ID XXX (XenD26411), xli) Sequence ID XXX (XenD26412), xlii) Sequence ID XXX (XenD26413),xliii) Sequence ID XXX (XenD26414), xliv) Sequence ID XXX (XenD26415), xlv) Sequence ID XXX (XenD26416), xlvi) Sequence ID XXX (XenD26417), xlvii) Sequence ID XXX (XenD26418), xlviii) Sequence ID XXX (XenD27070), xlix) Sequence ID XXX (XenD27071), l) Sequence ID XXX (XenD27072), li) Sequence ID XXX (XenD27073), lii) Sequence ID XXX (XenD27074), liii) Sequence ID XXX (X enD27075), liv) Sequence ID XXX (XenD27076), lv) Sequence ID XXX (XenD27077), lvi) Sequence ID XXX (XenD27078), lvii) Sequence ID XXX (XenD28173), lviii) Sequence ID XXX (XenD24876), lix) Sequence ID XXX (XenD27162), lx) Sequence ID XXX (XenD27163), lxi) Sequence ID XXX (XenD27164), lxii) Sequence ID XXX (XenD27165), lxiii) Sequence ID XXX (XENP31582, lxiv) Distribution, Column number XXX (XENP31583 chain 1), lxv) Sequence number XXX (XENP31584 chain 1), lxvi) Sequence number XXX (XENP32187 chain 1), lxvii) Sequence number XXX (XENP32188 chain 1), lxviii) Sequence number XXX (XENP32189, chain 1), lxix) Sequence number XXX (XENP32190 chain 1), lxx) Sequence number XXX (XENP32191 chain 1), lxxi) Sequence number XXX (XENP32991 chain 1), l xxii) Sequence ID XXX (XENP32992 chain 1), lxxiii) Sequence ID XXX (XENP32993 chain 1), lxxiv) Sequence ID XXX (XENP32994 chain 1), lxxv) Sequence ID XXX (XENP32995 chain 1), lxxvi) Sequence ID XXX (XENP32996 chain 1), lxxvi) Sequence ID XXX (XENP32997 chain 1), lxxvii) Sequence ID XXX (XENP32998 chain 1), lxxvii) Sequence ID XXX (XE NP32999 (chain 1), lxxviii) Sequence ID XXX (XENP33000 chain 1), lxxix) Sequence ID XXX (XENP33001 chain 1), lxxx) Sequence ID XXX (XENP33002 chain 1), lxxxi) Sequence ID XXX (XENP33003 chain 1), lxxxii) Sequence ID XXX (XENP33004 chain 1), lxxxiii) Sequence ID XXX (XENP33005 chain 1), lxxxiv) Sequence ID XXX (XENP33006 chain 1), The first fusion protein having a polypeptide sequence selected from the group consisting of lxxxv) SEQ ID NO: XXX (XENP33007 chain 1), lxxxvi) SEQ ID NO: XXX (XENP33008 chain 1), lxxxvii) SEQ ID NO: XXX (XENP33008 chain 1), lxxxviii) SEQ ID NO: XXX (XENP33009 chain 1), lxxxix) SEQ ID NO: XXX (XENP33010 chain 1), and xc) SEQ ID NO: XXX (XENP33011 chain 1),

[0022] b)i) Sequence ID 48 (XENP27201 Chain 2), ii) Sequence ID 126 (XenD24779), iii) Sequence ID 127 (XenD24780), iv) Sequence ID 128 (XenD24781), v) Sequence ID 129 (XenD24782), vi) Sequence ID 130 (XenD24783), vii) Sequence ID 131 (XenD24784), viiii) Sequence ID 132 (XenD24785), ix) Sequence ID 133 (XenD24786), x) Sequence ID 134 (XenD24787), xi) Sequence ID 135 (XenD24788), xii) Sequence IDs 136 (XenD24789), xiii) 137 (XenD24790), xiv) 138 (XenD24791), xv) 236 (XenD25911), xvi) 237 (XenD25912), xvii) 238 (XenD25913), xviii) 239 (XenD25914), xix) 240 (XenD25915), xx) 241 (XenD25916), xxi) 242 (XenD25917), xxii) 243 (XenD25918) xxiii) Sequence ID 244 (XenD25919), xxiv) Sequence ID 245 (XenD25920), xxv) Sequence ID 246 (XenD25921), xxvi) Sequence ID XXX (XenD26427), xxvii) Sequence ID XXX (XenD26428), xxviii) Sequence ID XXX (XenD27089), xxix) Sequence ID XXX (XenD27090), xxx) Sequence ID XXX (XenD27091), xxxxi) Sequence ID XXX (XenD27092), xxxii) Sequence ID XXX (XenD27093), xxxii i) Sequence ID XXX (XenD27094), xxxix) Sequence ID XXX (XenD24877), xl) Sequence ID XXX (XenD27166), xli) Sequence ID XXX (XENP31582, chain 2), xlii) Sequence ID XXX (XENP31583, chain 2), xliii) Sequence ID XXX (XENP31584, chain 2), xliv) Sequence ID XXX (XENP32187, chain 2), xlv) Sequence ID XXX (XENP32188, chain 2), xlvi) Sequence ID XXX (XENP32189, chain 2), xlvii) Sequence ID XXX (XENP32190, chain 2),xlviii) Sequence ID XXX (XENP32191 chain 2), xlix) Sequence ID XXX (XENP32991 chain 2), l) Sequence ID XXX (XENP32992 chain 2), li) Sequence ID XXX (XENP32993 chain 2), lii) Sequence ID XXX (XENP3, 2994 (chain 2), liii) Sequence ID XXX (XENP32995, liv) Sequence ID XXX (XENP32996, lv) Sequence ID XXX (XENP32997, lv) Sequence ID XXX (XENP32998, lvii) Sequence ID XXX (XENP32999, lviii) Sequence ID XXX (XENP33000, lvix) Sequence ID XXX (XENP33001, lvix) Sequence ID XXX (XENP33002, lvix) Sequence ID XXX (XENP33003, lvixii) Sequence ID XXX (XENP33004, lvixiii) The present invention provides a heterodimer Fc fusion protein comprising a second fusion protein having a polypeptide sequence selected from the group consisting of sequence number XXX (XENP33005 chain 2), lxiv) SEQ ID NO: XXX (XENP33006 chain 2), lxv) SEQ ID NO: XXX (XENP33007 chain 2), lxvi) SEQ ID NO: XXX (XENP33008 chain 2), lxvii) SEQ ID NO: XXX (XENP33008 chain 2), lxviii) SEQ ID NO: XXX (XENP33009 chain 2), lxix) SEQ ID NO: XXX (XENP33010 chain 2), and lxx) SEQ ID NO: XXX (XENP33011 chain 2).

[0023] In some embodiments, the present invention provides heterodimer Fc fusion proteins, such as XENP27201, XENP28820, XENP28821, XENP28822, XENP28823, XENP28824, XENP28825, XENP28826, XENP28827, XENP28828, XENP28829, XENP28830, XENP28831, XENP28832, XENP28833, XENP28834, XENP28835, XENP28836, XENP28837, XEN P28838, XENP28839, XENP28840, XENP28841, XENP28842, XENP28843, XENP28844, XENP28845, XENP28846, XENP28847, XENP28848, XENP28849, XENP288 50.XENP28851, NP30603, XENP30604, XENP30605, XENP30606, XENP30307, XENP30308, XENP30609, XENP31250, XENP31251, XENP31252, XENP31253, XENP31254, XENP31 255, XENP31256, XENP31257, XENP31258, XENP31259, XENP31260, XENP31261, XENP31262, XENP31263,XENP31264, ENP31143, XENP31144, XENP31145, XENP31146, XENP31582, XENP31583, XENP31584, XENP32186, XENP32187, XENP32188, XENP32189, XENP32190, XENP3 2191, XENP32991, XENP32992, XENP32993, XENP32994, XENP32995, XENP32996, XENP32997, XENP32998, XENP32999, XENP33000,The following are selected from the group consisting of XENP33003, XENP33004, XENP33005, XENP33006, XENP33007, XENP33008, XENP33008, XENP33009, XENP33010, and XENP33011.

[0024] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the heterodimer Fc fusion protein is selected from the group consisting of XENP31251, XENP31254, XENP31258, XENP32186, XENP32187, XENP32188, XENP32189, XENP32190, and XENP32191.

[0025] In some embodiments, the present invention provides a heterodimer Fc fusion protein further comprising one or more modifications to an IL-12p40 subunit selected from the group consisting of N103Q, N113Q, N200Q, and N281Q.

[0026] In some embodiments, the present invention provides compositions comprising heterodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0027] In some embodiments, the present invention provides one or more nucleic acids encoding heterodimeric Fc fusion proteins.

[0028] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a heterodimeric Fc fusion protein.

[0029] In some embodiments, the present invention provides a method for producing a heterodimer Fc fusion protein, the method comprising culturing host cells under conditions that produce the heterodimer Fc fusion protein.

[0030] In some embodiments, the present invention provides a method for purifying a heterodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a heterodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the heterodimeric Fc fusion protein.

[0031] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F 60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N1 03, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D16 6, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204 , F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0032] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0033] In another aspect, the present invention relates to a) a first fusion protein comprising a first protein domain and a first Fc domain, wherein the first protein domain comprises the first Fc The present invention provides a heterodimerized Fc fusion protein comprising: a) a first fusion protein covalently bonded to the C-terminus of a domain; and b) a second fusion protein comprising a second protein domain and a second Fc domain, wherein the second protein domain is covalently bonded to the C-terminus of the Fc domain, and the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain comprising an IL-12p40 subunit, and the second protein domain comprising an IL-12p35 subunit.

[0034] In some embodiments, the modifications that promote heterodimerization of the first and second Fc domains are a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including cross-linked disulfides, T366S / L368A / Y407V / Y349C:T366W / S354C), according to EU numbering, to provide a heterodimer Fc fusion protein.

[0035] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which a first protein domain is bound to a first Fc domain using a first domain linker and / or a second protein domain is bound to a second Fc domain using a second domain linker.

[0036] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0037] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0038] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0039] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0040] In some embodiments, the present invention is such that the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit This provides a heterodimer Fc fusion protein in which the variant IL-12p35 subunit is located.

[0041] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0042] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F 60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N1 03, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D16 6, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204 , F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0043] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E The present invention provides a heterodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0044] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q , S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K2 60E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144 K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S , E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N2 00D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N11 3Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N1 03Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59 K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K2 64E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200 Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N The present invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of 200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0045] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q , S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99 E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E , E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E 59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N1 03D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N 103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113 Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252 S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E , E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E 59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59 The present invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0046] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45 Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E 45Q / Q56E), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p 40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K),xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 2 08(IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209(IL-12p40(S43E / E59Q)), xliv) Sequence ID 210(IL-12p40(S43K / E49Q)), xlv) Sequence ID 211(IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212(IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213(IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214(IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX(IL-12p40(D34N / E59K)), l) Sequence ID XXX(IL-12p40, (E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL- 12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX(IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX(IL-12p40(C252S)), lxii) Sequence ID XXX(IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N1 03D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)),lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), l xxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX(IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX(IL-12p40(E32Q / D34N / E59 K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / C252S / K264E)),c) Sequence ID XXX(IL-12p40(E59K / K99E / , N281Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX (IL-1 2p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264 The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of E)), cvi) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) SEQ ID NO: XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)). In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0047] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0048] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0049] In some embodiments, the present invention relates to the IL-12p35 subunit, i) Sequence ID 113 (IL-12p35(N71D)), ii) Sequence ID 114 (IL-12p35(N85D)), iii) Sequence ID 115 (IL-12p35(N195D)), iv) Sequence ID 116 (IL-12p35(N71D / N85D / N195D)), v) Sequence Number 117 (IL-12p35(E153Q)), vi) Sequence ID 118 (IL-12p35(E38Q)), vii) Sequence ID 119 (IL-12p35(N151D)), viiii) Sequence ID 120 (IL-12p35(Q135E)), ix) Sequence ID 121 (IL-12p35(Q35D)), x) Sequence ID 122 (IL-12p35(Q146E )), xi) Sequence ID 123 (IL-12p35(N76D)), xii) Sequence ID 124 (IL-12p35(E162Q)), xiii) Sequence ID 125 (IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX (IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), xviii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227 (IL-12p35(N151K)), x xi) Sequence ID 226 (IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225 (IL-12p35(N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35(N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N1 51D), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence Number XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)),This provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of xxxix) SEQ ID NO: XXX(IL-12p35(N71Q / N195Q)), xl) SEQ ID NO: XXX(IL-12p35(N85Q / N195Q), xli) SEQ ID NO: XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii) SEQ ID NO: XXX(IL-12p35(N71D / N85D)), xliii) SEQ ID NO: XXX(IL-12p35(N71D / N195D), xliv) SEQ ID NO: XXX(IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX(IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX(IL-12p35(D55K)).

[0050] In some embodiments, the present invention provides a heterodimer Fc fusion protein, the heterodimer Fc fusion protein comprising a) a first fusion protein having the polypeptide sequence of SEQ ID NO: 49 (XENP27202 chain 1), and b) a second fusion protein having the polypeptide sequence of SEQ ID NO: 50 (XENP27202 chain 2).

[0051] In some embodiments, the present invention provides a heterodimer Fc fusion protein, said heterodimer Fc fusion protein being XENP27202.

[0052] In some embodiments, the present invention provides compositions comprising heterodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0053] In some embodiments, the present invention provides one or more nucleic acids encoding heterodimeric Fc fusion proteins.

[0054] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a heterodimeric Fc fusion protein.

[0055] In some embodiments, the present invention provides a method for producing a heterodimer Fc fusion protein, the method comprising culturing host cells under conditions that produce the heterodimer Fc fusion protein.

[0056] In some embodiments, the present invention provides a method for purifying a heterodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a heterodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the heterodimeric Fc fusion protein.

[0057] In another aspect, the present invention provides a heterodimer Fc fusion protein comprising: a) a fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is covalently bonded to the second protein domain, and the second protein domain is covalently bonded to the N-terminus of the first Fc domain; and b) a second Fc domain comprising a second Fc domain, wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain comprising an IL-12p40 subunit, and the second protein domain comprising an IL-12p35 subunit.

[0058] In some embodiments, the modifications that promote heterodimerization of the first and second Fc domains are a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including cross-linked disulfides, T366S / L368A / Y407V / Y349C:T366W / S354C), according to EU numbering, to provide a heterodimer Fc fusion protein.

[0059] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first protein domain is bound to the second protein domain using a first domain linker and / or the second protein domain is bound to the first Fc domain using a second domain linker.

[0060] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0061] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0062] In some embodiments, the present invention comprises the IL-12p40 subunit, which consists of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence). The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group, and / or the IL-12p35 subunit having a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0063] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0064] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0065] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0066] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F 60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N1 03, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D16 6, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204 , F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of F247, S248, Q256, K158, C252, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0067] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E The present invention provides a heterodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0068] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E5 9K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K This invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0069] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p4 0(Q56E)), ix) Sequence ID 65(IL-12p40(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67(IL-12p40(Q42E / E45Q)), xii) Sequence ID 68(IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70(IL-12p40(Q56E / E59Q)), xv) Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72(IL-12p40(E45Q / Q56E / E59Q)) xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 (IL-12p40(Q229E)), xxv) Sequence ID 81 (IL-12p 40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxx) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K2 64E), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q) ), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)),l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S) ), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N1 03D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)),lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi), Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxx iii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxx ix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / C252S / K264E)),c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K26 4E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N2 81Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX (IL-1 This provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of 2p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)) and cix)Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0070] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q1 Heterozygous, having one or more amino acid modifications as amino acid residues selected from the group consisting of 30, Q135, N136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196. This provides a dimeric Fc fusion protein.

[0071] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0072] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0073] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvi, i) Sequence ID XXX(IL-12p35(N195Q)), xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX(IL-12p35(N85Q / N195Q), xli) Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii) Sequence ID XXX(IL-12p The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of 35(N71D / N85D)), xliii)Sequence ID XXX(IL-12p35(N71D / N195D), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)).

[0074] In some embodiments, the present invention provides a heterodimer Fc fusion protein comprising a) the fusion protein having the polypeptide sequence of SEQ ID NO: 51 (XENP27203 chain 1), and b) the second Fc domain having the polypeptide sequence of SEQ ID NO: 52 (XENP27203 chain 2).

[0075] In some embodiments, the present invention provides a heterodimer Fc fusion protein, said heterodimer Fc fusion protein being XENP27203.

[0076] In some embodiments, the present invention provides a heterodimer Fc fusion protein, said heterodimer Fc fusion protein being XENP31290.

[0077] In some embodiments, the present invention provides compositions comprising heterodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0078] In some embodiments, the present invention provides one or more nucleic acids encoding heterodimeric Fc fusion proteins.

[0079] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a heterodimeric Fc fusion protein.

[0080] In some embodiments, the present invention provides a method for producing a heterodimer Fc fusion protein, the method comprising culturing host cells under conditions that produce the heterodimer Fc fusion protein.

[0081] In some embodiments, the present invention provides a method for purifying a heterodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a heterodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the heterodimeric Fc fusion protein.

[0082] In another aspect, the present invention provides a heterodimer Fc fusion protein comprising: a) a fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is covalently bonded to the second protein domain and the second protein domain is covalently bonded to the N-terminus of the first Fc domain; and b) a second Fc domain, wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain comprises an IL-12p35 subunit, and the second protein domain comprises an IL-12p40 subunit.

[0083] In some embodiments, heterodimerization of the first and second Fc domains is performed. The present invention provides a heterodimeric Fc fusion protein in which the promoting modification is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including cross-linked disulfides, T366S / L368A / Y407V / Y349C:T366W / S354C), according to EU numbering.

[0084] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first protein domain is bound to the second protein domain using a first domain linker and / or the second protein domain is bound to the first Fc domain using a second domain linker.

[0085] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0086] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0087] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and / or SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0088] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0089] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0090] In some embodiments, the present invention is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), The present invention provides a heterodimeric Fc fusion protein which is a variant IL-12p35 subunit having altered affinity for the IL-12 receptor complex.

[0091] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F 60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N1 03, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D16 6, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204 , F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0092] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E The present invention provides a heterodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0093] In some embodiments, the present invention relates to the IL-12p40 subunit, which is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D 18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144 K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E5 9K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N1 03Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K 264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N 281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C25 2S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K 264E, E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q The present invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0094] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45 Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E 45Q / Q56E), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p 40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K),xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K2 60E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), ,xlvi)Sequence ID 212(IL-12p40(E59Q / K99E)),xlvii)Sequence ID 213(IL-12p40(E59Q / K258E)),xlviii)Sequence ID 214(IL-12p40(E59Q / K260E)),xlix)Sequence ID XXX(IL-12p40(D34N / E59K)),l)Sequence ID XXX(IL-12p40(E59K / K99E)),li)Sequence ID XXX(IL-12p40(D18K / E59K / K99E)),lii)Sequence ID XXX(IL-12p40(E59K / K99E / K264E) ), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40( E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)),lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxvi ii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv ) Sequence ID XXX(IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX(IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), x ci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)),xcvii) Sequence ID XXX (IL-12p40(E59K / , K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99 E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of / C252S / K264E)), cviii)Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix)Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0095] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0096] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0097] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0098] In some embodiments, the present invention relates to the IL-12p35 subunit, i) Sequence ID 113 (IL-12p35(N71D)), ii) Sequence ID 114 (IL-12p3 5(N85D)), iii) Sequence ID 115(IL-12p35(N195D)), iv) Sequence ID 116(IL-12p35(N71D / N85D / N195D)), v) Sequence ID 117(IL-12p35(E153Q)), vi) Sequence ID 118(IL-12p35(E38Q)), vii) Sequence ID 119(IL-12p35(N151D)), viiii) Sequence ID 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), xviii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL- 12p35(E153K)), xxii)IL-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 228(IL-12p35(N151D / D165N)), xxvi)Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii)Sequence ID 230(IL-12p35(N151D / K158E)), xxviii)Sequence ID 231(IL-12p35(E79Q / N151D)), xxix)Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233(IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234(IL-12p35(N21D / N151D)), xxxii) Sequence ID 235(IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX(IL-12p35(F96A)), xxxiv) Sequence ID XXX(IL-12p35(M97A)), xxxxv) Sequence ID XXX(IL-12p35(L89A)), xxxvi) Sequence ID XXX(IL-12p35(L124A)),xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX This invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of (IL-12p35(N85Q / N195Q), xli)Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii)Sequence ID XXX(IL-12p35(N71D / N85D)), xliii)Sequence ID XXX(IL-12p35(N71D / N195D), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)).

[0099] In some embodiments, the present invention provides a heterodimer Fc fusion protein comprising a) the fusion protein having the polypeptide sequence of SEQ ID NO: 53 (XENP27204 chain 1), and b) the second Fc domain having the polypeptide sequence of SEQ ID NO: 54 (XENP27204 chain 2).

[0100] In some embodiments, the present invention provides a heterodimer Fc fusion protein, said heterodimer Fc fusion protein being XENP27204.

[0101] In some embodiments, the present invention provides compositions comprising heterodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0102] In some embodiments, the present invention provides one or more nucleic acids encoding heterodimeric Fc fusion proteins.

[0103] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a heterodimeric Fc fusion protein.

[0104] In some embodiments, the present invention provides a method for producing a heterodimer Fc fusion protein, the method comprising culturing host cells under conditions that produce the heterodimer Fc fusion protein.

[0105] In some embodiments, the present invention provides a method for purifying a heterodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a heterodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the heterodimeric Fc fusion protein.

[0106] In another aspect, the present invention provides a heterodimerized Fc fusion protein comprising: a) a fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is bound to the C-terminus of the first Fc domain and the second protein domain is covalently bound to the first protein domain; and b) a second Fc domain, wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain comprises an IL-12p40 subunit, and the second protein domain comprises an IL-12p35 subunit.

[0107] In some embodiments, the modifications that promote heterodimerization of the first and second Fc domains are a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including cross-linked disulfides, T366S / L368A / Y407V / Y349C:T366W / S354C), according to EU numbering, to provide a heterodimer Fc fusion protein.

[0108] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first protein domain is bound to the second protein domain using a first domain linker and / or the second protein domain is bound to the first Fc domain using a second domain linker.

[0109] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0110] In some embodiments, the present invention provides an amino acid substitution in which the first and / or second Fc domain is selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering. This provides a heterodimeric Fc fusion protein having an additional set.

[0111] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0112] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0113] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0114] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0115] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F 60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N1 03, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D16 6, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204 , F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0116] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K163E, E187Q, N200D, N200Q, N2 The present invention provides a heterodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of 18Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0117] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E5 9K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K This invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0118] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p4 0(N103D / N113D / N200D / N281D)), vi) Sequence ID 62(IL-12p40(Q42E)), vii) Sequence ID 63(IL-12p40(E45Q)), viiii) Sequence ID 64(IL-12p40(Q56E)), ix) Sequence ID 65(IL-12p40(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 ( IL-12p40(E73Q)), xix) Sequence ID 75(IL-12p40(Q144E)), xx) Sequence ID 76(IL-12p40(E262Q)), xxi) Sequence ID 77(IL-12p40(E100Q)), xxii) Sequence ID 78(IL-12p40(D18N)), xxiii) Sequence ID 79(IL-12p40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), x xvii) Sequence ID 83 (IL-12p40(E299Q)), xxviii) Sequence ID 84 (IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xx xv)IL-12p40(K99E), xxxvi)IL-12p40(K163E), xxxvii)IL-12p40(E187Q), xxxviii)IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii)Sequence ID 209(IL-12p40(S43E / E59Q)),xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence number XXX(IL-12p40(D18K / E59K / K99E)), lii) XXX(IL-12p40(E59K / K99E / K264E)), liii) XXX(IL-12p40(E59K / K99Y)), liv) XXX(IL-12p40(E59Y / K99E)), lv) XXX(IL-12p40(E45K / E59K / K99E)), lvi) XXX(IL-12p40(E59K / K99E / Q144E)), lvii) XXX(IL-12p40(E59K / K99E / Q 144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252 S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)),lxx) Sequence number XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence number, Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID X XX(IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX(IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX(IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID X XX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX(IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX(IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX(IL-12p40(E32K / D34N / E59K / K99E)),xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL -12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E) ), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N1 This provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of 03Q / N113Q / N200Q / C252S / K264E), cviii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0119] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0120] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0121] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0122] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX(IL-12p35(M97A)), xxxv) Sequence ID XXX(IL-12p, 35(L89A)), xxxvi) Sequence ID XXX(IL-12p35(L124A)), xxxvii) Sequence ID XXX(IL-12p35(M125A)), xxxviii) Sequence ID XXX(IL-12p35(L75A)), xxxiv) Sequence ID XXX(IL-12p35(I171A)), xxxv) Sequence ID XXX(IL-12p35(N71Q)), xxxvi) Sequence ID XXX(IL-12p35(N85Q)), xxxvii) Sequence ID XXX(IL-12p35(N195Q)), xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71 The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of Q / N195Q), xl) SEQ ID NO: XXX (IL-12p35(N85Q / N195Q), xli) SEQ ID NO: XXX (IL-12p35(N71Q / N85Q / N195Q)), xlii) SEQ ID NO: XXX (IL-12p35(N71D / N85D)), xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D), xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0123] In some embodiments, the present invention provides compositions comprising heterodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0124] In some embodiments, the present invention provides one or more nucleic acids encoding heterodimeric Fc fusion proteins.

[0125] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a heterodimeric Fc fusion protein.

[0126] In some embodiments, the present invention provides a method for producing a heterodimer Fc fusion protein, the method comprising culturing host cells under conditions that produce the heterodimer Fc fusion protein.

[0127] In some embodiments, the present invention provides a method for purifying a heterodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a heterodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the heterodimeric Fc fusion protein.

[0128] In another aspect, the present invention provides a heterodimerized Fc fusion protein comprising: a) a fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is bound to the C-terminus of the first Fc domain and the second protein domain is covalently bound to the first protein domain; and b) a second Fc domain, wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain comprises an IL-12p35 subunit, and the second protein domain comprises an IL-12p40 subunit.

[0129] In some embodiments, the modifications that promote heterodimerization of the first and second Fc domains are from the group consisting of L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including cross-linked disulfides, T366S / L368A / Y407V / Y349C:T366W / S354C) or The present invention provides a heterodimeric Fc fusion protein, which is a set of amino acid substitutions selected from the above.

[0130] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first protein domain is bound to the second protein domain using a first domain linker and / or the second protein domain is bound to the first Fc domain using a second domain linker.

[0131] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0132] In some embodiments, the present invention provides a heterodimeric Fc fusion protein in which the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0133] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0134] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0135] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0136] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0137] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L4 7, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E18 The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 7, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0138] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E The present invention provides a heterodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0139] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N, 103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59 The present invention provides a heterodimer Fc fusion protein having amino acid substitutions selected from the group consisting of K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0140] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45 Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E 45Q / Q56E), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p 40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K),xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187 Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209(I L-12p40(S43E / E59Q)), xliv) Sequence ID 210(IL-12p40(S43K / E49Q)), xlv) Sequence ID 211(IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212(IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213(IL-12p40(E59Q / K258E)), xlviii ) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E, / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL -12p40(E59K / K99E / K264E)), lx) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX(IL-12p40(C252S)), lxii) Sequence ID XXX(IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX(IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX(I L-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX(IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX(IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX(IL-12p40(N103D / N200D)), lxix) Sequence ID XXX(IL-12p40(N103D / N281D)), lxx) Sequence ID XXX(IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX(IL-12p40(N113D / N 281D), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)),lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi ) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)),ci) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii), Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q The present invention provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0141] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a heterodimer Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0142] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0143] In some embodiments, the present invention provides a heterodimer Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0144] In some embodiments, the present invention relates to the IL-12p35 subunit, i) Sequence ID 113 (IL-12p35(N71D)), ii) Sequence ID 114 (IL-12p35(N85D)), iii) Sequence ID 115 (IL-12p35(N195D)), iv) Sequence ID 116 (IL-12p35(N71D / N85D / N195D)), v) Sequence Number 117 (IL-12p35(E153Q)), vi) Sequence ID 118 (IL-12p35(E38Q)), vii) Sequence ID 119 (IL-12p35(N151D)), viiii) Sequence ID 120 (IL-12p35(Q135E)), ix) Sequence ID 121 (IL-12p35(Q35D)), x) Sequence ID 122 (IL-12p35(Q146E )), xi) Sequence ID 123 (IL-12p35(N76D)), xii) Sequence ID 124 (IL-12p35(E162Q)), xiii) Sequence ID 125 (IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX (IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), xviii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227 (IL-12p35(N151K)), x xi) Sequence ID 226 (IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225 (IL-12p35(N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35(N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N1 51D), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence Number XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)),This provides a heterodimer Fc fusion protein having a polypeptide sequence selected from the group consisting of xxxix) SEQ ID NO: XXX(IL-12p35(N71Q / N195Q)), xl) SEQ ID NO: XXX(IL-12p35(N85Q / N195Q), xli) SEQ ID NO: XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii) SEQ ID NO: XXX(IL-12p35(N71D / N85D)), xliii) SEQ ID NO: XXX(IL-12p35(N71D / N195D), xliv) SEQ ID NO: XXX(IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX(IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX(IL-12p35(D55K)).

[0145] In some embodiments, the present invention provides compositions comprising heterodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0146] In some embodiments, the present invention provides one or more nucleic acids encoding heterodimeric Fc fusion proteins.

[0147] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a heterodimeric Fc fusion protein.

[0148] In some embodiments, the present invention provides a method for producing a heterodimer Fc fusion protein, the method comprising culturing host cells under conditions that produce the heterodimer Fc fusion protein.

[0149] In some embodiments, the present invention provides a method for purifying a heterodimer Fc fusion protein, the method comprising: a) providing a composition comprising a heterodimer Fc fusion protein; b) packing the composition into an ion exchange column; and c) the heterodimer Fc This includes collecting fractions containing fusion proteins.

[0150] In another embodiment, the present invention provides a heterodimer complex comprising a) an IL-12p40 subunit and b) an IL-12p35 subunit.

[0151] In some embodiments, the present invention provides a heterodimer complex in which the IL-12p40 subunit is covalently bonded to the IL-12p35 subunit using a domain linker.

[0152] In some embodiments, the present invention provides a heterodimer complex in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0153] In some embodiments, the present invention provides a heterodimer complex in which the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

[0154] In some embodiments, the present invention provides a heterodimer complex in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0155] In some embodiments, the present invention provides a heterodimer complex in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0156] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59 , F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102 , N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y The present invention provides a heterodimer complex having one or more amino acid modifications at amino acid residues selected from the group consisting of 246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0157] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q1 The present invention provides a heterodimer complex having one or more amino acid substitutions selected from the group consisting of 44E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0158] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E The present invention provides a heterodimer complex having amino acid substitutions selected from the group consisting of 59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0159] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p 40(E45Q / Q56E)), xiii) Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70(IL-12p40(Q56E / E59Q)), xv) Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72(IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73(IL-12p40(D161N)), xviii) Sequence ID 74(IL-12p40(E73Q)), xix) Sequence ID 75(IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 (IL-12p40(Q229E)), xxv) Sequence ID 81 (IL-12p40(E235Q)), xxvi) Sequence ID 82 (IL-12p40(Q256N)), xxvii) Sequence ID 83 (IL-12p40(E299Q)), xxviii) Sequence ID 84 (IL-12p40(D87N)) , xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence number XXX(IL-12p40(E59K / K99E / N103Q / N11) 3Q / N200Q / N281Q / C252S / K264E)), xxxiv) SEQ ID NO: 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q),xxxviii)IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii)Sequence ID 209(IL-12p40(S43E / E59Q)), xliv)Sequence ID 210(IL-12p40(S43K / E49Q)), xlv)Sequence ID 211(IL-12p40(E59Q / K 163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Column number XXX(IL-12p40(E59K / K99E / K264E)), lx) Sequence number XXX(IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence number XXX(IL-12p40(C252S)), lxii) Sequence number XXX(IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence number XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence number XXX(IL-12p40(E59K / K99Y / C252S)),lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K2, 64E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID X XX(IL-12p40(N113Q)), lxxix) Sequence ID XXX(IL-12p40(N200Q)), lxxx) Sequence ID XXX(IL-12p40(N281Q)), lxxxi) Sequence ID XXX(IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX(IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX(IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX(IL-12p40(N113Q / N200Q)), lxxxv) Sequence Number XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)),xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12 p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)),The present invention provides a heterodimer complex having a polypeptide sequence selected from the group consisting of and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0160] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q13 The present invention provides a heterodimer complex having one or more amino acid modifications as amino acid residues selected from the group consisting of 5, N136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0161] In some embodiments, the present invention provides a heterodimer complex in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0162] In some embodiments, the present invention provides a heterodimer complex in which the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0163] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35 (N151D)), viii) Sequence ID 120 (IL-12p35(Q135E)), ix) Sequence ID 121 (IL-12p35(Q35D)), x) Sequence ID 122 (IL-12p35(Q146E)), xi) Sequence ID 123 (IL-12p35(N76D)), xii) Sequence ID 124 (IL-12p35(E162Q)), xiii) Sequence ID 125 (IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX (IL-12p35(D55Q)), xvi) I L-12p35(E79Q), xvii)IL-12p35(Q130E), xviii)IL-12p35(N136D), xix)IL-12p35(E143Q), xx)Sequence ID 227(IL-12p35(N151K)), xxi)Sequence ID 226(IL-12p35(E153K)), xxii)IL-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 228(IL-12 p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230(IL-12p35(N151D / K158E)), xxviii) Sequence ID 231(IL-12p35(E79Q / N151D)), xxix) Sequence ID 232(IL-12p35(D55Q / N151D)), xxx) Sequence ID 233(IL-12p35(N136D / N151D)), xxxx) Sequence ID 234(IL-12p35(N21D / N151D )), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID X XX(IL-12p35(M125A)), xxxviii) Sequence ID XXX(IL-12p35(L75A)), xxxiv) Sequence ID XXX(IL-12p35(I171A)), xxxv) Sequence ID XXX(IL-12p35(N71Q)), xxxvi) Sequence ID XXX(IL-12p35(N85Q)), xxxvii) Sequence ID XXX(IL-12p35(N195Q)), xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX(IL-12p35(N85Q / N195Q), xli) Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii) Sequence ID XXX(IL-12p35(N71D / N8 The present invention provides a heterodimer complex having a polypeptide sequence selected from the group consisting of 5D)), xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D), xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0164] In some embodiments, the present invention provides compositions comprising heterodimer complexes for use in the treatment of cancer in a subject.

[0165] In some embodiments, the present invention provides one or more nucleic acids that encode heterodimer complexes.

[0166] In some embodiments, the present invention provides a host cell containing one or more nucleic acids that encode a heterodimer complex.

[0167] In some embodiments, the present invention provides a method for producing a heterodimer complex, the method comprising culturing host cells under conditions that produce the heterodimer complex.

[0168] In some embodiments, the present invention provides a method for purifying a heterodimer complex, the method comprising: a) providing a composition comprising the heterodimer complex; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the heterodimer complex.

[0169] In another embodiment, the present invention provides IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or variant IL-12p40 subunits having modified affinity for the IL-12 receptor complex.

[0170] In some embodiments, the present invention provides a variant IL-12p40 subunit, which is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q 56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102 , N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170 It has one or more amino acid modifications with amino acid residues selected from the group consisting of Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, Q256, C252, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299. In some embodiments, the present invention provides a variant IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103 It has one or more amino acid substitutions selected from the group consisting of D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0171] In some embodiments, the present invention provides a variant IL-12p40 subunit, which is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D3 4N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E5 9Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E5 9K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59 K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K26 4E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C 252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N2 00D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113 D / N200D, N103D / N113D / N281D, N103D / N200D / N281D, N113D / N200D / N281D, N 103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N20 0Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E,E59K / K99E / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N 200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K 264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K9 9E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K, It has amino acid substitutions selected from the group consisting of 99E / N113Q / N200Q / N281Q / C252S / K264E and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0172] In some embodiments, the present invention provides variant IL-12p40 subunits, which include i) SEQ ID NO: 57 (IL-12p40(N103D)), ii) SEQ ID NO: 58 (IL-12p40(N113D)), iii) SEQ ID NO: 59 (IL-12p40(N200D)), iv) SEQ ID NO: 60 (IL-12p40(N281D)), v) SEQ ID NO: 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) SEQ ID NO: 62 (IL-12p40(Q42E)), v ii) Sequence ID 63 (IL-12p40(E45Q)), viii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 ( IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72(IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73(IL-12p40(D161N)), xviii) Sequence ID 74(IL-12p40(E73Q)), xix) Sequence ID 75(IL-12p40(Q144E)), xx) Sequence ID 76(IL-12p40(E262Q)), xxi) Sequence ID 77(IL-12p40(E100Q)), xxii) Sequence ID 78(IL-12p40(D18N)), xxiii) Sequence ID 7 9(IL-12p40(E33Q)), xxiv)Sequence ID 80(IL-12p40(Q229E)), xxv)Sequence ID 81(IL-12p40(E235Q)), xxvi)Sequence ID 82(IL-12p40(Q256N)), xxvii)Sequence ID 83(IL-12p40(E299Q)), xxviii)Sequence ID 84(IL-12p40(D87N)), xxix)IL-12p40(E32Q), xxx)IL-12p40(D34N), xxxxi)IL-12p40(S43E), xxxii)IL-12p40(S43K),xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-1 2p40(E59Q / K99E)), xlvii) Sequence ID 213(IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214(IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX(IL-12p40(D34N / E59K)), l) Sequence ID XXX(IL-12p40(E59K / K99E)), li) Sequence ID XXX(IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12, p40(E59K / K99E / R159E)), lix) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX(IL-12p40(C252S)), lxii) Sequence ID XXX(IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX(IL- 12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX(IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX(IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX(IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX(IL-12p40(N103D / N200D)), lxix) Sequence ID XXX(IL-12p40(N103D / N281D)), lxx) Sequence ID XXX(IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N 281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)),lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N2 81Q)), xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D 34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)),cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C, The polypeptide sequence is selected from the group consisting of 252S / K264E), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0173] In some embodiments, the present invention provides compositions comprising the variant IL-12p40 subunit.

[0174] In some embodiments, the present invention provides nucleic acids encoding the variant IL-12p40 subunit.

[0175] In some embodiments, the present invention provides a host cell containing the nucleic acid encoding the variant IL-12p40 subunit.

[0176] In some embodiments, the present invention provides a method for producing a variant IL-12p40 subunit, the method comprising culturing the host cells described in claim H8 under conditions that produce the variant IL-12p40 subunit.

[0177] In another embodiment, the present invention provides IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or variant IL-12p35 subunits having modified affinity for the IL-12 receptor complex.

[0178] In some embodiments, the present invention provides a variant IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, It has one or more amino acid modifications as amino acid residues selected from the group consisting of M97, L124, M125, Q130, Q135, N136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0179] In some embodiments, the present invention provides a variant IL-12p35 subunit having one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0180] In some embodiments, the present invention provides variant IL-12p35 subunits, which are N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q The amino acid substitutions are selected from the group consisting of / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0181] In some embodiments, the present invention provides a variant IL-12p35 subunit having the amino acid substitutions N71D / N85D / N195D.

[0182] In some embodiments, the present invention provides a variant IL-12p35 subunit, which is i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 1 19(IL-12p35(N151D)), viii) Sequence ID 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p3 5(E79Q), xvii)IL-12p35(Q130E), xviii)IL-12p35(N136D), xix)IL-12p35(E143Q), xx)Sequence ID 227(IL-12p35(N151K)), xxi)Sequence ID 226(IL-12p35(E153K)), xxii)IL-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxx) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)),xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID The polypeptide sequence is selected from the group consisting of XXX(IL-12p35(N71Q / N85Q)), xxxix)Sequence ID XXX(IL-12p35(N71Q / N195Q)), xl)Sequence ID XXX(IL-12p35(N85Q / N195Q)), xli)Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii)Sequence ID XXX(IL-12p35(N71D / N85D)), xliii)Sequence ID XXX(IL-12p35(N71D / N195D)), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)).

[0183] In some embodiments, the present invention provides compositions comprising the variant IL-12p35 subunit.

[0184] In some embodiments, the present invention provides nucleic acids encoding the variant IL-12p35 subunit.

[0185] In some embodiments, the present invention provides a host cell containing the nucleic acid encoding the variant IL-12p35 subunit.

[0186] In some embodiments, the present invention provides a variant IL-12p35 subunit, the method comprising culturing host cells under conditions in which the variant IL-12p35 subunit is produced.

[0187] In another embodiment, the present invention comprises a) a first fusion protein comprising an IL-12p40 subunit domain covalently bound to a first Fc domain, and b) a second fusion protein comprising an IL-12p35 subunit domain covalently bound to a second Fc domain, wherein the first and second Fc domains are designated according to EU numbering as L368D / K370S and S364K;L368D / K370S and S364K / E357L;L368D / The present invention provides heterodimeric proteins that are sets of amino acid substitutions selected from the group consisting of K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q; T366S / L368A / Y407V and T366W; and T366S / L368A / Y407V / Y349C and T366W / S354C;.

[0188] In some embodiments, the present invention provides a heterodimer protein in which the IL-12p40 subunit domain is bound to the N-terminus of the first Fc domain and the IL-12p35 subunit domain is bound to the N-terminus of the second Fc domain.

[0189] In some embodiments, the present invention provides a heterodimer protein in which the IL-12p40 subunit domain is bound to the C-terminus of the first Fc domain and the IL-12p35 subunit domain is bound to the C-terminus of the second Fc domain.

[0190] In some embodiments, the present invention provides a heterodimer protein in which the Il-12p40 subunit domain is bound to the first Fc domain using a domain linker, and the IL-12p35 subunit domain is bound to the second Fc domain using a domain linker.

[0191] In some embodiments, the present invention provides a heterodimer protein in which the first and second Fc domains comprise IgG1 hinge-CH2-CH3.

[0192] In some embodiments, the present invention provides a heterodimer protein in which the first fusion protein comprises an IL-12p40 subunit domain-domain linker-hinge-CH2-CH3 from the N-terminus to the C-terminus, and the second fusion protein comprises an IL-12p35 subunit domain-domain linker-hinge-CH2-CH3 from the N-terminus to the C-terminus.

[0193] In some embodiments, the present invention relates to a first fusion protein having a hinge-CH2-CH3-domain linker-IL-12p40 subunit domain from the N-terminus to the C-terminus. The present invention provides a heterodimer protein comprising a second fusion protein containing a hinge-CH2-CH3-domain linker-IL-12p35 subunit domain from the N-terminus to the C-terminus.

[0194] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45 Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E 45Q / Q56E), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p 40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K),xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206( IL-12p40(E32Q / E59Q)), xli) Sequence ID 207(IL-12p40(D34N / E59Q)), xlii) Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209(IL-12p40(S43E / E59Q)), xliv) Sequence ID 210(IL-12p40(S43K / E49Q)), xlv) Sequence ID 211(IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212(IL-12p40(E59Q / K99E) ), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID number Sequence number XXX (IL-12p40(E59K / K99Y)), liv) Sequence number XXX (IL-12p40(E59Y / K99E)), lv) Sequence number XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence number XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence number XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence number XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence number, Number XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N11 3D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q )), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)),lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q) )), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y )), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)),cvi) Sequence ID XXX(IL-1, The present invention provides a heterodimer protein having a sequence selected from the group consisting of 2p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0195] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E)), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85 The present invention provides a heterodimer protein having a sequence selected from the group consisting of Q)), xxxix) SEQ ID NO: XXX(IL-12p35(N71Q / N195Q)), lx) SEQ ID NO: XXX(IL-12p35(N85Q / N195Q)), lxi) SEQ ID NO: XXX(IL-12p35(N71Q / N85Q / N195Q)), lxii) SEQ ID NO: XXX(IL-12p35(N71D / N85D)), lxiii) SEQ ID NO: XXX(IL-12p35(N71D / N195D)), lxiv) SEQ ID NO: XXX(IL-12p35(N85D / N195D)), lxv) SEQ ID NO: XXX(IL-12p35(D55Q)), and lxvi) SEQ ID NO: XXX(IL-12p35(D55K)).

[0196] In another embodiment, the present invention comprises a heterodimer protein comprising a) a first Fc domain and a fusion protein comprising b) i) a second Fc domain, ii) an IL-12p40 subunit domain, and iii) an IL-12p35 subunit domain, wherein the first and second Fc domains are designated according to EU numbering L368D / K370S and S364K; L368D / K370S and S364K / E357 This is a set of amino acid substitutions selected from the group consisting of L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q;T366S / L368A / Y407V and T366W; and T366S / L368A / Y407V / Y349C and T366W / S354C;.

[0197] In some embodiments, the present invention provides a heterodimer protein comprising an IL-12p40 subunit domain - domain linker - IL-12p35 subunit domain - domain linker - hinge - CH2 - CH3 from the N-terminus to the C-terminus.

[0198] In some embodiments, the present invention provides a heterodimer protein comprising an IL-12p35 subunit domain-domain linker-IL-12p40 subunit domain-domain linker-hinge-CH2-CH3 from the N-terminus to the C-terminus.

[0199] In some embodiments, the present invention provides a heterodimer protein comprising a hinge-CH2-CH3-domain linker-IL-12p35 subunit domain-domain linker-IL-12p40 subunit domain from the N-terminus to the C-terminus.

[0200] In some embodiments, the present invention provides a heterodimer protein comprising a hinge-CH2-CH3-domain linker-IL-12p40 subunit domain-domain linker-IL-12p35 subunit domain from the N-terminus to the C-terminus.

[0201] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi )Sequence ID 62(IL-12p40(Q42E)), vii)Sequence ID 63(IL-12p40(E45Q)), viiii)Sequence ID 64(IL-12p40(Q56E)), ix)Sequence ID 65(IL-12p40(E59Q)), x)Sequence ID 66(IL-12p40(D62N)), xi)Sequence ID 67(IL-12p40(Q42E / E45Q)), xii)Sequence ID 68(IL-12p40(E45Q / Q56E xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p4 0(E73Q)), xix) Sequence ID 75(IL-12p40(Q144E)), xx) Sequence ID 76(IL-12p40(E262Q)), xxi) Sequence ID 77(IL-12p40(E100Q)), xxii) Sequence ID 78(IL-12p40(D18N)), xxiii) Sequence ID 79(IL-12p40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(I L-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q) ), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX(IL-12p40(E59K / K99E)), li) Sequence ID XXX(IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX(IL-12p40(E59K / K99Y)), liv) Sequence ID XXX(IL-12p40(E59Y / K99E)), lv) Sequence ID XXX(IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX(IL-12p40(E59K / K99E / Q144E)),lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii ) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX(IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX(IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX(IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX(IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX(IL-12p40(N103D / N20 0D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Column number XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence number XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence number XXX (IL-12p40(N103Q)), lxxviii) Sequence number XXX (IL-12p40(N113Q)), lxxix) Sequence number XXX (IL-12p40(N200Q)), lxxx) Sequence number XXX (IL-12p40(N281Q)), lxxxi) Sequence number XXX (IL-12p40(N103Q / N113Q)),lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lx, xxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281 Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E3 2Q / D34N / E59K / K99E)), xcv) Sequence ID XXX(IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N2 c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)),civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99 This invention provides a heterodimer protein having a sequence selected from the group consisting of E / N103Q / N113Q / N200Q / C252S / K264E), cviii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0202] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120 (IL-12p35(Q135E)), ix) SEQ ID NO: 121 (IL-12p35(Q35D)), x) Sequence ID: Sequence number 122 (IL-12p35(Q146E)), xii) Sequence number 123 (IL-12p35(N76D)), xii) Sequence number 124 (IL-12p35(E162Q)), xiii) Sequence number 125 (IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence number XXX (IL-12p35(D5 5Q)), xvi)IL-12p35(E79Q), xvii)IL-12p35(Q130E), xviii)IL-12p35(N136D), xix)IL-12p35(E143Q), xx)Sequence ID 227(IL-12p35(N151K)), xxi)Sequence ID 226(IL-12p35(E153K)), xxii)I L-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 228(IL-12p35(N151D / D165N)), xxvi)Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii)Sequence ID 230(IL-12p35(N151D / K158E)), xxviii)Sequence ID 231(IL-12p35(E79Q / N151D)), xxix)Sequence ID 232(IL-12p35(D55Q / N15 1D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX ( IL-12p35(M125A)), xxxviii) Sequence ID XXX(IL-12p35(L75A)), xxxiv) Sequence ID XXX(IL-12p35(I171A)), xxxv) Sequence ID XXX(IL-12p35(N71Q)), xxxvi) Sequence ID XXX(IL-12p35(N85Q)), xxxvii) Sequence ID XXX(IL-12p35(N195Q)), xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71Q / N195Q)), lx) The sequence has sequences selected from the group consisting of sequence number XXX (IL-12p35(N85Q / N195Q), lxi) sequence number XXX (IL-12p35(N71Q / N85Q / N195Q)), lxii) sequence number XXX (IL-12p35(N71D / N85D)), lxiii) sequence number XXX (IL-12p35(N71D / N195D), lxiv) sequence number XXX (IL-12p35(N85D / N195D)), lxv) sequence number XXX (IL-12p35(D55Q)), and lxvi) sequence number XXX (IL-12p35(D55K)),It provides heterodimer proteins.

[0203] In another embodiment, the present invention provides a homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from N-terminus to C-terminus, an IL-12p40 subunit domain - an optional first domain linker - an IL-12p35 subunit domain - an optional second domain linker - an Fc domain.

[0204] In some embodiments, the present invention provides a homodimerized Fc fusion protein in which the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C, according to EU numbering.

[0205] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0206] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0207] In some embodiments, the present invention is defined as the Fc domain being G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E The present invention provides a homodimeric Fc fusion protein having an additional set of amino acid substitutions selected from the group consisting of 233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K.

[0208] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: XX (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: XX (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: XX (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: XX (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0209] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0210] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0211] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0212] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G 61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D 170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F 206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F2 The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 47, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0213] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0214] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E 59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59 This invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0215] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)) ,viii)Sequence ID 64(IL-12p40(Q56E)),ix)Sequence ID 65(IL-12p40(E59Q)),x)Sequence ID 66(IL-12p40(D62N)),xi)Sequence ID 67(IL-12p40(Q42E / E45Q)),xii)Sequence ID 68(IL-12p40(E45Q / Q56E)),xiii)Sequence ID 69(IL-12p40(Q42E / E59Q)),xiv)Sequence ID 70(IL-12p40(Q56E / E59Q)),xv)Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)),x vi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 ( IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX(IL-12p40(E59K / K99 E / N103Q / N113Q / N200Q / N281Q / C252S / K264E), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)),xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (I L-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX(IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX(IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX(IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX(IL-12p40(N103D / N113D)),lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID X XX(IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX(IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX(IL-12p40(N103Q)), lxxviii) Sequence ID XXX(IL-12p40(N113Q)), lxxix) Sequence ID XXX(IL-12p40(N200Q)), lxxx) Sequence ID XXX(IL-12p40(N281Q)), lxxxi) Sequence ID XXX(IL-12p40(N103Q / N113Q)), lxxxi i) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX(IL-12p40(D34K / E59K / K99E)),xciv) Sequence ID XXX(IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX(IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)) , xcix) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XX This provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of X(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii)Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix)Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0216] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, The present invention provides a homodimer Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0217] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0218] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0219] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 12 0(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), xviii )IL-12p35(N136D), xix)IL-12p35(E143Q), xx)Sequence ID 227(IL-12p35(N151K)), xxi)Sequence ID 226(IL-12p35(E153K)), xxii)IL-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 228(IL-12p35(N151D / D165N)), xxvi)Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxx) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence number Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), xxxix The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of ) SEQ ID NO: XXX (IL-12p35(N71Q / N195Q)), xl) SEQ ID NO: XXX (IL-12p35(N85Q / N195Q)), xli) SEQ ID NO: XXX (IL-12p35(N71Q / N85Q / N195Q)), xlii) SEQ ID NO: XXX (IL-12p35(N71D / N85D)), xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D)), xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0220] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0221] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0222] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0223] In some embodiments, the present invention provides one or more nucleic acids encoding homodimeric Fc fusion proteins.

[0224] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0225] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing host cells under conditions that produce the homodimeric Fc fusion protein.

[0226] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a homodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the homodimeric Fc fusion protein.

[0227] In another embodiment, the present invention provides a homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from N-terminus to C-terminus, an IL-12p35 subunit domain - an optional first domain linker - an IL-12p40 subunit domain - an optional second domain linker - an Fc domain.

[0228] In some embodiments, the present invention relates to a modification that promotes homodimerization of the Fc domain, according to EU numbering, L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T We provide a homodimeric Fc fusion protein which is a set of amino acid substitutions selected from the group consisting of 366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C.

[0229] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0230] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0231] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0232] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0233] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0234] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0235] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0236] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73 , K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159 , D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0237] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0238] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N1 03Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C, The present invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of 252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E. In some embodiments, the present invention provides the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL -12p40(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67(IL-12p40(Q42E / E45Q)), xii) Sequence ID 68(IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70(IL-12p40(Q56E / E59Q)), xv) Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72(IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 (IL-12p40(Q229E)), xxv) Sequence ID 81 (IL-12p40(E23 5Q), xxvi) Sequence ID 82 (IL-12p40(Q256N)), xxvii) Sequence ID 83 (IL-12p40(E299Q)), xxviii) Sequence ID 84 (IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xl i) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Column number 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence number 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence number 214 (IL-12p40(E59Q / K260E)), xlix) Sequence number XXX (IL-12p40(D34N / E59K)), l) Sequence number XXX (IL-12p40(E59K / K99E)), li) Sequence number XX X(IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX(IL-12p40(E59K / K99Y)), liv) Sequence ID XXX(IL-12p40(E59Y / K99E)), lv) Sequence ID XXX(IL-12p40(E45K / E59K / K99E)), lvi, ) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX ( IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX(IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX(IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX(IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX(IL-12p40(N103D / N113 D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XX X(IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX(IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX(IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX(IL-12p40(N103Q)), lxxviii) Sequence ID XXX(IL-12p40(N113Q)), lxxix) Sequence ID XXX(IL-12p40(N200Q)), lxxx) Sequence ID XXX(IL-12p40(N281Q)),lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E )), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX (IL-12p40(E 59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)),ciii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Distribution, Column number XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence number XXX(IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence number XXX(IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence number XXX(IL-12p40(E59K / K99E / N103Q The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of / N113Q / N200Q / C252S / K264E)), cviii)Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix)Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0239] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0240] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0241] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0242] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 11 9(IL-12p35(N151D)), viii) Sequence ID 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(I L-12p35(D55Q)), xvi)IL-12p35(E79Q), xvii)IL-12p35(Q130E), xviii)IL-12p35(N136D), xix)IL-12p35(E143Q), xx)Sequence ID 227(IL-12p35(N151K)), xxi)Sequence ID 226(IL-12p35(E153K)), xxii)IL-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 2 28(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230(IL-12p35(N151D / K158E)), xxviii) Sequence ID 231(IL-12p35(E79Q / N151D)), xxix) Sequence ID 232(IL-12p35(D55Q / N151D)), xxx) Sequence ID 233(IL-12p35(N136D / N151D)), xxxx) Sequence ID 234(IL-12p35(N21D / N151D)), xx xii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35( I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35(N85Q / N195Q), xli) Sequence ID XXX (IL-12p35(N71Q / N85Q / N195Q)),Provide a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of: (xlii) SEQ ID NO: XXX (IL-12p35 (N71D / N85D)), (xliii) SEQ ID NO: XXX (IL-12p35 (N71D / N195D)), (xliv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), (xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and (xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

[0243] In some embodiments, provide a homodimeric Fc fusion protein wherein the first and second monomers are XENP31289.

[0244] In some embodiments, provide a homodimeric Fc fusion protein wherein the first and second monomers are XENP31291.

[0245] In some embodiments, the present invention provides a homodimeric Fc fusion protein composition comprising a homodimeric Fc fusion protein for use in the treatment of cancer in a subject.

[0246] In some embodiments, the present invention provides one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0247] In some embodiments, the present invention provides a host cell comprising the one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0248] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing a host cell under conditions in which the homodimeric Fc fusion protein is produced.

[0249] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising the homodimeric Fc fusion protein; b) loading the composition onto an ion exchange column; and c) collecting the fraction containing the homodimeric Fc fusion protein.

[0250] In another aspect, the present invention provides a homodimeric Fc fusion protein comprising a first monomer and a second monomer, each comprising, from N-terminus to C-terminus, an Fc domain - an optional first domain linker - an IL-12p40 subunit domain - an optional second domain linker - an IL-12p35 subunit domain.

[0251] In some embodiments, the present invention provides a homodimeric Fc fusion protein wherein the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S; S364K; S364K / E357L; S364K / E357Q; T411E / K360E / Q362E; D401K; T366S / L368A / Y407V; T366W; T366S / L368A / Y407V / Y349C; and T366W / S354C according to EU numbering.

[0252] In some embodiments, the present invention provides a homodimeric Fc fusion protein wherein the first domain linker and the second domain linker have the same amino acid sequence.

[0253] In some embodiments, the present invention provides a homodimeric Fc fusion protein wherein the Fc domain has an additional set of amino acid substitutions comprising Q295E / N384D / Q418E / N421D according to EU numbering.

[0254] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0255] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0256] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0257] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0258] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0259] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G 61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D 170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F 206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F2 The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 47, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0260] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0261] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K2 60E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K2 64E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E5 9K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N200D , N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N200D / N281D , N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C 252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E, E59K / K9 9E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K9 9E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K9 The present invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of 9E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0262] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45 Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E 45Q / Q56E), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p 40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K),xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii), IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii)Sequence ID 209(IL-12p40(S43E / E59Q)), xliv)Sequence ID 210(IL-12p40(S43K / E49Q)), xlv)Sequence ID 211(IL-12p40(E59Q / K163E)) ,xlvi)Sequence code 212(IL-12p40(E59Q / K99E)),xlvii)Sequence code 213(IL-12p40(E59Q / K258E)),xlviii)Sequence code 214(IL-12p40(E59Q / K260E)),xlix)Sequence code XXX(IL-12p40(D34N / E59K)),l)Sequence code XXX(IL-12p40(E59K / K99E)),li)Sequence code XXX(IL-12p40(D18K / E59K / K99E)),lii)Sequence code XXX(IL-12p40(E59K / K99E) / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)),lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID X XX(IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX(IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX(IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX(IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX(IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX(IL-12p40(N113D / N200D / N281D)), lxxvii ) Sequence ID XXX(IL-12p40(N103Q)), lxxviii) Sequence ID XXX(IL-12p40(N113Q)), lxxix) Sequence ID XXX(IL-12p40(N200Q)), lxxx) Sequence ID XXX(IL-12p40(N281Q)), lxxxi) Sequence ID XXX(IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX(IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX(IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX(IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX(IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX(IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)),xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N2, 00Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59 K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C25 The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of 2S / K264E), cvii)Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii)Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix)Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0263] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0264] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0265] In some embodiments, the present invention relates to the IL-12p35 subunit being N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D. This invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the following group.

[0266] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), x This provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of xxix) SEQ ID NO: XXX (IL-12p35(N71Q / N195Q)), xl) SEQ ID NO: XXX (IL-12p35(N85Q / N195Q)), xli) SEQ ID NO: XXX (IL-12p35(N71Q / N85Q / N195Q)), xlii) SEQ ID NO: XXX (IL-12p35(N71D / N85D)), xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D)), xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0267] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0268] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0269] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0270] In some embodiments, the present invention provides one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0271] In some embodiments, the present invention provides a host cell comprising the one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0272] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing a host cell under conditions in which the homodimeric Fc fusion protein is produced.

[0273] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising the homodimeric Fc fusion protein; b) loading the composition onto an ion exchange column; and c) collecting the fraction containing the homodimeric Fc fusion protein.

[0274] In another aspect, the present invention provides a homodimeric Fc fusion protein comprising a first monomer and a second monomer, each comprising, from N-terminus to C-terminus, an Fc domain - optional first domain linker - IL-12p35 subunit domain - optional second domain linker - IL-12p40 subunit domain.

[0275] In some embodiments, the present invention provides a homodimeric Fc fusion protein, wherein the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S; S364K; S364K / E357L; S364K / E357Q; T411E / K360E / Q362E; D401K; T366S / L368A / Y407V; T366W; T366S / L368A / Y407V / Y349C; and T366W / S354C according to EU numbering.

[0276] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0277] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0278] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0279] In some embodiments, the present invention relates to the IL-12p40 subunit, and the sequence number is 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and sequence number The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of Sequence ID No. 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), wherein the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of Sequence ID No. 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and Sequence ID No. 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0280] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0281] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0282] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0283] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G 61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D 170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F 206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F2 The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 47, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0284] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0285] In some embodiments, the present invention relates to the IL-12p40 subunit, N103D / N113D / N200D / N281D、Q42E / E45Q、E45Q / Q56E、Q42E / E59Q、Q56E / E59Q、Q4 2E / E45Q / Q56E、E45Q / Q56E / E59Q、E32Q / E59Q、D34N / E59K、D34N / E59K / K99E 、D34K / E59K / K99E、E32Q / D34N / E59K / K99E、E32K / D34N / E59K / K99E、D34N / E 59Q、E59Q / E187Q、S43E / E59Q、S43K / E49Q、E59Q / K163E、E59Q / K99E、E59Q / K 258E、E59Q / K260E、E59K / K99E、D18K / E59K / K99E、E59K / K99E / K264E、E59K / K99Y、E59Y / K99Y、E59Y / K99E、E45K / E59K / K99E、E59K / K99E / Q144E、E59K / K 99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252 S、E59K / K99E / C252S / K264E、E59K / K99E / C252S、N103D / N113D、N103D / N200 D、N103D / N281D、N113D / N200D、N113D / N281D、N200D / N281D、N103D / N113D / N200D、N103D / N113D / N281D、N103D / N200D / N281D、N113D / N200D / N281D、N1 03Q / N113Q、N103Q / N200Q、N103Q / N281Q、N113Q / N200Q、N113Q / N281Q、N200Q / N281Q、N103Q / N113Q / N200Q、N103Q / N113Q / N281Q、N103Q / N200Q / N281Q、N 113Q / N200Q / N281Q、N103Q / N113Q / N200Q / N281Q、E59K / K99E / N103Q / C252S / K264E、E59K / K99E / N113Q / C252S / K264E、E59K / K99E / N200Q / C252S / K264E、E59K / K99E / N281Q / C252S / K264E、E59K / K99E / N103Q / N113Q / C252S / K264E、E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113 This invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

[0286] In some embodiments, the present invention relates to the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (I L-12p40(E59Q)), x) Sequence ID 66 (IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74(IL-12p40(E73Q)), xix) Sequence ID 75(IL-12p40(Q144E)), xx) Sequence ID 76(IL-12p40(E262Q)), xxi) Sequence ID 77(IL-12p40(E100Q)), xxii) Sequence ID 78(IL-12p40(D18N)), xxiii) Sequence ID 79(IL-12p40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N) xxvii) Sequence ID 83 (IL-12p40(E299Q)), xxviii) Sequence ID 84 (IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40(E59K)) xxxv)IL-12p40(K99E), xxxvi)IL-12p40(K163E), xxxvii)IL-12p40(E187Q), xxxviii)IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii)Sequence ID 209(IL-12p40(S43E / E59 Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)),li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K 99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)),lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxx, vii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Column number XXX(IL-12p40(N113Q / N200Q)), lxxxv) Sequence number XXX(IL-12p40(N113Q / N281Q)), lxxxvi) Sequence number XXX(IL-12p40(N200Q / N281Q)), lxxxvii) Sequence number XXX(IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence number XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence number XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Array Number XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E )), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)),ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence This provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of sequence number XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0287] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0288] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0289] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0290] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX (IL, The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of -12p35(N71Q / N195Q)), xl)Sequence ID XXX(IL-12p35(N85Q / N195Q), xli)Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii)Sequence ID XXX(IL-12p35(N71D / N85D)), xliii)Sequence ID XXX(IL-12p35(N71D / N195D), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)).

[0291] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0292] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0293] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0294] In some embodiments, the present invention provides one or more nucleic acids encoding homodimeric Fc fusion proteins.

[0295] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0296] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing host cells under conditions that produce the homodimeric Fc fusion protein.

[0297] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a homodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the homodimeric Fc fusion protein.

[0298] In another embodiment, the present invention provides a homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising an IL-12p40 subunit domain - first domain linker - IL-12p35 subunit domain - second domain linker - Fc domain from the N-terminus to the C-terminus.

[0299] In some embodiments, the present invention provides a homodimerized Fc fusion protein in which the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C, according to EU numbering.

[0300] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0301] In some embodiments, the present invention relates that the Fc domain is assigned the EU number Q29 The present invention provides a homodimeric Fc fusion protein having an additional set of amino acid substitutions, including 5E / N384D / Q418E / N421D.

[0302] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0303] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0304] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0305] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0306] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0307] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R15 9, D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E18 1, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q22 Select from the group consisting of 9, E231, E235, T242, P243, S245, Y246, F247, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299. The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at the amino acid residues being treated.

[0308] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0309] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E 59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59 This invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E. In some embodiments, the present invention provides the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-1 2p40(N113D)), iii) Sequence ID 59(IL-12p40(N200D)), iv) Sequence ID 60(IL-12p40(N281D)), v) Sequence ID 61(IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62(IL-12p40(Q42E)), vii) Sequence ID 63(IL-12p40(E45Q)), viiii) Sequence ID 64(IL-12p40(Q56E)), ix) Sequence ID 65(IL-12p40(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67 (IL-12p40(Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69 (IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40(Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40 (E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 (IL-12p40(Q229E)), xxv) Sequence ID 81 (IL-12p40(E235Q)), xxvi) Sequence ID 82 (IL-12p40(Q256N)), xxvii) Sequence ID 83 (I L-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205(IL-12p40(E59K)), xxxv) IL-12p40(K99E),xxxvi)IL-12p40(K163E), xxxvii)IL-12p40(E187Q), xxxviii)IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii)Sequence ID 209(IL-12p40(S43E / E59Q)), xliv)Sequence ID 210(IL-1 2p40(S43K / E49Q)), xlv) Sequence ID 211(IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212(IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213(IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214(IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX(IL-12p40(D34N / E59K)), l) Sequence ID XXX(IL-12p40(E59K / K99E)), li) Sequence ID XXX(IL-12p40(D 18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii ) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)),lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C25, 2S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N103D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Column number XXX (IL-12p40(N113D / N281D)), lxxii) Sequence number XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence number XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence number XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence number XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence number XXX (IL-12p40(N113D / N200D / N281D)), lxx vii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxi v) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)),xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C 252S / K264E)), c) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)),and cix) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)), This invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the following group.

[0310] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0311] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0312] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0313] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p 35(E38Q)), vii) Sequence ID 119(IL-12p35(N151D)), viiii) Sequence ID 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35 (E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), xviii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) I L-12p35(K158E), xxiii)IL-12p35(D165N), xxiv)Sequence ID 225(IL-12p35(N151D / E153Q)), xxv)Sequence ID 228(IL-12p35(N151D / D165N)), xxvi)Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii)Sequence ID 230(IL-12p35(N151D / K158E)), xxviii)Sequence ID 231(IL-1 2p35(E79Q / N151D)), xxix) Sequence ID 232(IL-12p35(D55Q / N151D)), xxx) Sequence ID 233(IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234(IL-12p35(N21D / N151D)), xxxii) Sequence ID 235(IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX(IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX(IL-12p35(L89A)), xxxvi) Sequence ID XXX(IL-12p35(L124A)), xxxvii) Sequence ID XXX(IL-12p35(M125A)), xxxviii) Sequence ID XXX(IL-12p35(L75A)), xxxiv) Sequence ID XXX(IL-12p35(I171A)), xxxv) Sequence ID XXX(IL-12p35(N71Q)), xxxv i) Sequence ID XXX(IL-12p35(N85Q)), xxxvii) Sequence ID XXX(IL-12p35(N195Q)), xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX(IL-12p35(N85Q / N195Q), xli) Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xli i)Sequence ID XXX(IL-12p35(N71D / N85D)), xliii)Sequence ID XXX(IL-12p35(N71D / N195D), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)) provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of i)Sequence ID XXX(IL-12p35(N71D / N85D), xliii)Sequence ID XXX(IL-12p35(N71D / N195D), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)).

[0314] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0315] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0316] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0317] In some embodiments, the present invention provides one or more nucleic acids encoding homodimeric Fc fusion proteins.

[0318] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0319] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing host cells under conditions that produce the homodimeric Fc fusion protein.

[0320] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a homodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the homodimeric Fc fusion protein.

[0321] In another embodiment, the present invention provides a homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising an IL-12p35 subunit domain - first domain linker - IL-12p40 subunit domain - second domain linker - Fc domain from the N-terminus to the C-terminus.

[0322] In some embodiments, the present invention provides a homodimerized Fc fusion protein in which the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C, according to EU numbering.

[0323] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0324] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0325] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0326] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0327] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0328] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0329] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0330] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G 61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D 170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F 206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F2 The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 47, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0331] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0332] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q , S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99 E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E , E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E 59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N1 03D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N 103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113 Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252 S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E , E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E 59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59 The present invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E. In some embodiments, the present invention provides the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL -12p40(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67(IL-12p40(Q42E / E45Q)), xii) Sequence ID 68(IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70(IL-12p40(Q56E / E59Q)), xv) Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72(IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 (IL-12p40(Q229E)), xxv) Sequence ID 81 (IL-12p40(E23 5Q), xxvi) Sequence ID 82 (IL-12p40(Q256N)), xxvii) Sequence ID 83 (IL-12p40(E299Q)), xxviii) Sequence ID 84 (IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40(S43E / E 59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)), l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18, K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX ( IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX(IL-12p40(C252S)), lxii) Sequence ID XXX(IL-12p40(DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX(IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX(IL -12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX(IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX(IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX(IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX(IL-12p40(N103D / N200D)), lxix) Sequence ID XXX(IL-12p40(N103D / N281D)), lxx) Sequence ID XXX(IL-12p40(N113D / N2 00D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)),lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence number Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (I L-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX(IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX(IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX(IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12, p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of -12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0333] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0334] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0335] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0336] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120 (IL-12p35(Q135E)), ix) SEQ ID NO: 121 ( IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), xviii) IL-12p35(N136D), xix)(IL-12 p35(E143Q)), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230(IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)), xxxiv) Sequence ID XXX (IL-12p35(M97A) ), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)),xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX(IL-12p35(N85Q / N195Q), xli) Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii) Sequence ID XXX(IL-12p35(N71D / N85D) The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of (xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D), (xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), (xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and (xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0337] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0338] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0339] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0340] In some embodiments, the present invention provides one or more nucleic acids encoding homodimeric Fc fusion proteins.

[0341] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0342] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing host cells under conditions that produce the homodimeric Fc fusion protein.

[0343] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a homodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the homodimeric Fc fusion protein.

[0344] In another embodiment, the present invention provides a homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an Fc domain - first domain linker - IL-12p40 subunit domain - second domain linker - IL-12p35 subunit domain.

[0345] In some embodiments, the present invention provides a homodimerized Fc fusion protein in which the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C, according to EU numbering.

[0346] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0347] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0348] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0349] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0350] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0351] In some embodiments, the present invention provides that the IL-12p40 subunit is a Varian The present invention provides a homodimer Fc fusion protein which is an IL-12p40 subunit and / or which is a variant IL-12p35 subunit.

[0352] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0353] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G 61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D 170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F 206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F2 The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 47, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0354] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0355] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q , S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K2 60E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144 K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S , E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N2 00D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N11 3Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N1 03Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59 K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K2 64E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200 Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N The present invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of 200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E. In some embodiments, the present invention provides the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p4 0(E59Q)), x) Sequence ID 66(IL-12p40(D62N)), xi) Sequence ID 67(IL-12p40(Q42E / E45Q)), xii) Sequence ID 68(IL-12p40(E45Q / Q56E)), xiii) Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv) Sequence ID 70(IL-12p40(Q56E / E59Q)), xv) Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)), xvi) Sequence ID 72(IL-12p40(E45Q / Q56E / E59Q)), xvii) Sequence ID 73(IL- 12p40(D161N)), xviii) Sequence ID 74(IL-12p40(E73Q)), xix) Sequence ID 75(IL-12p40(Q144E)), xx) Sequence ID 76(IL-12p40(E262Q)), xxi) Sequence ID 77(IL-12p40(E100Q)), xxii) Sequence ID 78(IL-12p40(D18N)), xxiii) Sequence ID 79(IL-12p40(E33Q)), xxiv) Sequence ID 80(IL-12p40(Q229E)), xxv) Sequence ID 81(IL-12p40(E235Q)), xxvi) Sequence ID 8 2(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205(IL-12.p40(E59K)), xxxv)IL-12p40(K99E), xxxvi)IL-12p40(K163E), xxxvii)IL-12p40(E187Q), xxxviii)IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliii)Sequence ID 209(IL -12p40(S43E / E59Q)), xliv) Sequence ID 210(IL-12p40(S43K / E49Q)), xlv) Sequence ID 211(IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212(IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213(IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214(IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX(IL-12p40(D34N / E59K)), l) Sequence ID XXX( IL-12p40(E59K / K99E)), li) Sequence ID XXX(IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX(IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX(IL-12p40(E59K / K99Y)), liv) Sequence ID XXX(IL-12p40(E59Y / K99E)), lv) Sequence ID XXX(IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX(IL-12p40(E59K / K99E / Q144E) ), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S)),lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N10 3D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N 113Q), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)),lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40(N103Q / N200Q, / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX(IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX(IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX(IL-12p40(E32 K / D34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX(IL-12p40(E59K / K99E / N28 1Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K264E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E) ), cv) Sequence ID XXX (IL-12p40(E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)),The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of and cix) SEQ ID NO: XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0356] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications as amino acid residues selected from the group consisting of 136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

[0357] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0358] In some embodiments, the present invention relates to the IL-12p35 subunit, which corresponds to N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151 The present invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0359] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvii) Sequence ID XXX (IL-12p35(N195Q)), xxxviii) Sequence ID XXX (IL-12p35(N71Q / N85Q)), x This provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of xxix) SEQ ID NO: XXX (IL-12p35(N71Q / N195Q)), xl) SEQ ID NO: XXX (IL-12p35(N85Q / N195Q)), xli) SEQ ID NO: XXX (IL-12p35(N71Q / N85Q / N195Q)), xlii) SEQ ID NO: XXX (IL-12p35(N71D / N85D)), xliii) SEQ ID NO: XXX (IL-12p35(N71D / N195D)), xliv) SEQ ID NO: XXX (IL-12p35(N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35(D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35(D55K)).

[0360] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0361] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0362] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0363] In some embodiments, the present invention provides one or more nucleic acids encoding homodimeric Fc fusion proteins.

[0364] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0365] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing host cells under conditions that produce the homodimeric Fc fusion protein.

[0366] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a homodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the homodimeric Fc fusion protein.

[0367] In another embodiment, the present invention provides a homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an Fc domain - first domain linker - IL-12p35 subunit domain - second domain linker - IL-12p40 subunit domain.

[0368] In some embodiments, the present invention provides a homodimerized Fc fusion protein in which the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C, according to EU numbering.

[0369] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the first domain linker and the second domain linker have the same amino acid sequence.

[0370] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

[0371] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_ / S267K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K according to EU numbering.

[0372] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

[0373] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which each of the Fc domains further comprises the amino acid substitution M428L / N424S.

[0374] In some embodiments, the present invention provides a homodimer Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

[0375] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

[0376] In some embodiments, the IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G 61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D 170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F 206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F2 The present invention provides a homodimeric Fc fusion protein having one or more amino acid modifications at amino acid residues selected from the group consisting of 47, S248, C252S, Q256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

[0377] In some embodiments, the present invention relates to the IL-12p40 subunit, which is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144 The present invention provides a homodimer Fc fusion protein having one or more amino acid substitutions selected from the group consisting of E, D161N, R159E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

[0378] In some embodiments, the present invention relates to the IL-12p40 subunit, which corresponds to N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E , D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E 45K / E59K / K99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E5 9K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N 113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N10 3D / N200D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N1 03Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E 59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59 This invention provides a homodimeric Fc fusion protein having amino acid substitutions selected from the group consisting of K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E. In some embodiments, the present invention provides the IL-12p40 subunit, i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65(IL-12p40(E59Q)), x)Sequence ID 66(IL-12p40(D62N)), xi)Sequence ID 67(IL-12p40(Q42E / E45Q)), xii)Sequence ID 68(IL-12p40(E45Q / Q56E)), xiii)Sequence ID 69(IL-12p40(Q42E / E59Q)), xiv)Sequence ID 70(IL-12p40(Q56E / E59Q)), xv)Sequence ID 71(IL-12p40(Q42E / E45Q / Q56E)), xvi)Sequence ID 72(IL-12p40(E45Q / Q56E / E5 9Q), xvii) Sequence ID 73 (IL-12p40(D161N)), xviii) Sequence ID 74 (IL-12p40(E73Q)), xix) Sequence ID 75 (IL-12p40(Q144E)), xx) Sequence ID 76 (IL-12p40(E262Q)), xxi) Sequence ID 77 (IL-12p40(E100Q)), xxii) Sequence ID 78 (IL-12p40(D18N)), xxiii) Sequence ID 79 (IL-12p40(E33Q)), xxiv) Sequence ID 80 (IL-12p40(Q229E)), xxv) Sequence ID 81 (IL- 12p40(E235Q)), xxvi) Sequence ID 82(IL-12p40(Q256N)), xxvii) Sequence ID 83(IL-12p40(E299Q)), xxviii) Sequence ID 84(IL-12p40(D87N)), xxix) IL-12p40(E32Q), xxx) IL-12p40(D34N), xxxxi) IL-12p40(S43E), xxxii) IL-12p40(S43K), xxxiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E), xxxiv) Sequence ID 205 (IL-12p40(E59K)), xxxv) IL-12p40(K99E), xxxvi) IL-12p40(K163E), xxxvii) IL-12p40(E187Q), xxxviii) IL-12p40(K258E), xxxix) IL-12p40(K260E), xl) Sequence ID 206 (IL-12p40(E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40(D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40(E59Q / E187Q) )), xliii) Sequence ID 209 (IL-12p40(S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40(S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40(E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40(E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40(E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40(E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40(D34N / E59K)),l) Sequence ID XXX (IL-12p40(E59K / K99E)), li) Sequence ID XXX (IL-12p40(D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40(E59K / K99Y)), liv) Sequence ID XXX (IL-12p40(E59Y / K99E)), lv) Sequence ID XXX (IL-12p40(E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40(E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40(E59K / K99E / Q144K)), lviii) Sequence ID XXX (IL-12p40(E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40(E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40(C252S)), lxii) Sequence ID XXX (IL-12p40(DI8K / E59K / K99E / C252S) ), lxiii) Sequence ID XXX (IL-12p40(D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40(E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40(E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40(E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40(N103D / N113D)), lxviii) Sequence ID XXX (IL-12p40(N1 03D / N200D)), lxix) Sequence ID XXX (IL-12p40(N103D / N281D)), lxx) Sequence ID XXX (IL-12p40(N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40(N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40(N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50(N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40(N103D / N113D / N281D)),lxxv) Sequence ID XXX (IL-12p40(N103D / N200D / N281D)), lx, xvi) Sequence ID XXX (IL-12p40(N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40(N103Q)), lxxviii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), l xxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N281Q)), lx xxix) Sequence ID XXX (IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40(D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40(D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40(E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX(IL-12p40(E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX(IL-12p40(E59Y / K99Y)), xcvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / C252S / K264E)), xcviii) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX(IL-12p40(E59K / K99E / N200Q / C252S / K264E)),c) Sequence ID XXX(IL-12p40(E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N200Q / C252S / K26 4E)), ciii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N 281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviii) Sequence ID XXX (IL- This provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of 12p40(E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)) and cix) Sequence ID XXX(IL-12p40(E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

[0379] In some embodiments, the present invention relates to the IL-12p35 subunit, which comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q1 Homoneutoglycan 30, Q135, N136, E143, Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196, having one or more amino acid modifications as amino acid residues selected from the group, This provides a mer-Fc fusion protein.

[0380] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

[0381] In some embodiments, the present invention provides a homodimeric Fc fusion protein in which the IL-12p35 subunit has amino acid substitutions selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

[0382] In some embodiments, the present invention relates to the IL-12p35 subunit, i) SEQ ID NO: 113 (IL-12p35(N71D)), ii) SEQ ID NO: 114 (IL-12p35(N85D)), iii) SEQ ID NO: 115 (IL-12p35(N195D)), iv) SEQ ID NO: 116 (IL-12p35(N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35(E153Q)), vi) SEQ ID NO: 118 (IL-12p35(E38Q)), vii) SEQ ID NO: 119 (IL-12p35(N151D)), viiii) SEQ ID NO: 120(IL-12p35(Q135E)), ix) Sequence ID 121(IL-12p35(Q35D)), x) Sequence ID 122(IL-12p35(Q146E)), xi) Sequence ID 123(IL-12p35(N76D)), xii) Sequence ID 124(IL-12p35(E162Q)), xiii) Sequence ID 125(IL-12p35(E163Q)), xiv) IL-12p35(N21D), xv) Sequence ID XXX(IL-12p35(D55Q)), xvi) IL-12p35(E79Q), xvii) IL-12p35(Q130E), x viii) IL-12p35(N136D), xix) IL-12p35(E143Q), xx) Sequence ID 227(IL-12p35(N151K)), xxi) Sequence ID 226(IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225(IL-12p35(N151D / E153Q)), xxv) Sequence ID 228(IL-12p35(N151D / D165N)), xxvi) Sequence ID 229(IL-12p35(Q130E / N151D)) xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35(D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35(N136D / N151D)), xxxxi) Sequence ID 234 (IL-12p35(N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35(E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35(F96A)),xxxiv) Sequence ID XXX (IL-12p35(M97A)), xxxv) Sequence ID XXX (IL-12p35(L89A)), xxxvi) Sequence ID XXX (IL-12p35(L124A)), xxxvii) Sequence ID XXX (IL-12p35(M125A)), xxxviii) Sequence ID XXX (IL-12p35(L75A)), xxxiv) Sequence ID XXX (IL-12p35(I171A)), xxxv) Sequence ID XXX (IL-12p35(N71Q)), xxxvi) Sequence ID XXX (IL-12p35(N85Q)), xxxvi, i) Sequence ID XXX(IL-12p35(N195Q)), xxxviii) Sequence ID XXX(IL-12p35(N71Q / N85Q)), xxxix) Sequence ID XXX(IL-12p35(N71Q / N195Q)), xl) Sequence ID XXX(IL-12p35(N85Q / N195Q), xli) Sequence ID XXX(IL-12p35(N71Q / N85Q / N195Q)), xlii) Sequence ID XXX(IL-12 The present invention provides a homodimeric Fc fusion protein having a polypeptide sequence selected from the group consisting of p35(N71D / N85D)), xliii)Sequence ID XXX(IL-12p35(N71D / N195D), xliv)Sequence ID XXX(IL-12p35(N85D / N195D)), xlv)Sequence ID XXX(IL-12p35(D55Q)), and xlvi)Sequence ID XXX(IL-12p35(D55K)).

[0383] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31289.

[0384] In some embodiments, a homodimeric Fc fusion protein is provided in which the first and second monomers are XENP31291.

[0385] In some embodiments, the present invention provides homodimeric Fc fusion protein compositions comprising homodimeric Fc fusion proteins for use in the treatment of cancer in a subject.

[0386] In some embodiments, the present invention provides one or more nucleic acids encoding homodimeric Fc fusion proteins.

[0387] In some embodiments, the present invention provides a host cell comprising one or more nucleic acids encoding a homodimeric Fc fusion protein.

[0388] In some embodiments, the present invention provides a method for producing a homodimeric Fc fusion protein, the method comprising culturing host cells under conditions that produce the homodimeric Fc fusion protein.

[0389] In some embodiments, the present invention provides a method for purifying a homodimeric Fc fusion protein, the method comprising: a) providing a composition comprising a homodimeric Fc fusion protein; b) packing the composition into an ion exchange column; and c) collecting a fraction containing the homodimeric Fc fusion protein.

[0390] In another embodiment, the present invention provides a method for treating cancer in a patient who requires treatment for cancer, comprising administering to the patient a therapeutically effective amount of a heterodimer or homodimer Fc fusion protein according to any of the preceding claims.

[0391] In some embodiments, the present invention provides a method for treating cancer in a patient who requires treatment for cancer, further comprising administering a therapeutically effective dose of a checkpoint-blocking antibody.

[0392] In some further embodiments, the present invention provides a method for treating cancer in a patient who requires treatment for cancer, wherein the checkpoint blocking antibody is selected from anti-PD-1 antibody, anti-PD-L1 antibody, anti-TIM3 antibody, anti-TIGIT antibody, anti-LAG3 antibody, and anti-CTLA-4 antibody.

[0393] In some further embodiments, the present invention provides a method for treating cancer in patients who require treatment for cancer, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab , or pizilizumab.

[0394] In some further embodiments, the present invention provides a method for treating cancer in a patient who requires treatment for cancer, wherein the anti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab.

[0395] In some further embodiments, the present invention provides a method for treating cancer in a patient who requires treatment for cancer, the patient exhibiting an increase in lymphocytes after administration.

[0396] In some further embodiments, the present invention provides a method for treating cancer in a patient who requires treatment for cancer, the patient exhibiting an increase in peripheral CD8+ T cells after administration.

[0397] In some embodiments, the present invention provides a method for treating cancer in patients who require treatment for cancer, wherein the IL-12Fc fusion protein is administered before a checkpoint inhibitor to increase pre-treatment checkpoint expression.

[0398] In some embodiments, the present invention provides a method for treating cancer in patients who require treatment for cancer, wherein the IL-12Fc fusion protein is administered before a checkpoint inhibitor to increase pre-treatment checkpoint expression.

[0399] In another embodiment, the present invention provides a method for inducing T cell proliferation in a patient who requires induction of T cell proliferation, comprising administering to the patient a therapeutically effective amount of a heterodimer or homodimer Fc fusion protein of any of the preceding items.

[0400] In some further embodiments, the present invention provides a method for inducing T cell proliferation in a patient who requires T cell proliferation, further comprising administering a therapeutically effective dose of a checkpoint-blocking antibody.

[0401] In some further embodiments, the present invention provides a method for inducing T cell proliferation in a patient who requires T cell proliferation, wherein the checkpoint blocking antibody is selected from anti-PD-1 antibody, anti-PD-L1 antibody, anti-TIM3 antibody, anti-TIGIT antibody, anti-LAG3 antibody, and anti-CTLA-4 antibody.

[0402] In some further embodiments, the present invention provides a method for inducing T cell proliferation in a patient who requires T cell proliferation, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, or pizilizumab.

[0403] In some other further embodiments, the present invention provides a method for inducing T cell proliferation in a patient who requires induction of T cell proliferation, wherein the anti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab.

[0404] In some further embodiments, the present invention provides a method for inducing T cell proliferation in a patient who requires T cell proliferation, wherein T cell proliferation is at least a twofold increase in T cells.

[0405] In some embodiments, the present invention provides a method for inducing T cell proliferation in patients who require T cell proliferation, wherein the IL-12Fc fusion protein is administered prior to a checkpoint inhibitor to increase pre-treatment checkpoint expression.

[0406] In some embodiments, the present invention relates to patients who require the induction of T cell proliferation. This method provides a way to induce T cell proliferation, and the IL-12Fc fusion protein is administered before checkpoint inhibitors to increase pre-treatment checkpoint expression.

[0407] In one embodiment, the present invention provides an IL-12p40 subunit. In another embodiment, the present invention provides an IL-12p35 subunit. In a further embodiment, the present invention provides a heterodimer complex comprising a) an IL-12p40 subunit and b) an IL-12p35 subunit. [Brief explanation of the drawing]

[0408] [Figure 1] Figures 1A and 1B show the sequences of IL-12 and its receptor. [Figure 2] Figures 2A–2E show useful pairs of Fc heterodimerization variants (including skew and pI variants). Variants that do not have a corresponding "monomer 2" are pI variants that can be used alone for any monomer. [Figure 3] A list of constant regions of isometric volume variant antibodies and their respective substitutions is shown. pI_(-) indicates a low pI variant, and pI_(+) indicates a high pI variant. These can be optionally and independently combined with other heterodimerized variants of the present invention (and other variant species outlined herein). [Figure 4] These compounds exhibit useful ablation variants that ablate the FcγR bond (sometimes referred to as "knockout" or "KO" variants). Generally, ablation variants are found in both monomers, but in some cases, they may be present in only one monomer. [Figure 5] Particularly useful embodiments of the "non-cytokine" component of the present invention are shown. [Figure 6]Several exemplary domain linkers are shown. In some embodiments, these linkers find use in binding an IL-12p35 subunit, an IL-12p40 subunit, or a single-chain IL-12 complex to the N-terminus of the Fc region. In some embodiments, these linkers find use in fusing an IL-12p35 subunit to an IL-12p40 subunit in a single-chain IL-12 complex. It is important to note that the scIL-12 complex can contain either IL-12p35 N-terminally bound to IL-12p40 or IL-12p40 N-terminally bound to IL-12p35. Also, in some cases described herein, the hinge portion of the Fc domain can function as a domain linker and can be combined with any of these linkers. [Figure 7]Figures 7A–7E show sequences of several useful IL-12-Fc fusion backbones based on human IgG1 that do not contain cytokine sequences (e.g., IL-12p35 subunit, IL-12p40 subunit, or scIL-12 complex). Backbone 1 is based on human IgG1 (356E / 358M allotype) and includes C220S, S364K / E357Q:L368D / K370S scuba riant on both strands, Q295E / N384D / Q418E / N421D pI variants on the strand with L368D / K370S scuba riant, and E233P / L234V / L235A / G236del / S267K ablation variants on both strands. Backbone 2 is based on human IgG1 (356E / 358M allotype) and includes C220S, S364K:L368D / K370S scuba riant on both strands, Q295E / N384D / Q418E / N421D pI variants on the strand with L368D / K370S scuba riant, and E233P / L234V / L235A / G236del / S267K ablation variants on both strands. Backbone 3 is based on human IgG1 (356E / 358M allotype) and includes C220S, S364K:L368E / K370S scuba riant on both strands, Q295E / N384D / Q418E / N421D pI variants on the strand with L368E / K370...

Claims

1. A heterodimer Fc fusion protein, a) A first fusion protein comprising a variant IL-12p40 subunit domain and a first Fc domain, wherein the IL-12p40 subunit domain is covalently bonded to the N-terminus of the first Fc domain, b) A second fusion protein comprising an IL-12p35 subunit domain and a second Fc domain, wherein the IL-12p35 subunit domain is covalently bonded to the N-terminus of the Fc domain, A heterodimerized Fc fusion protein wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains.

2. The IL-12p40 subunits are E59K, E59Q, D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R15 The heterodimer Fc fusion protein according to claim 1, having one or more amino acid substitutions selected from the group consisting of 9E, K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

3. The IL-12p40 subunit is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E5 9Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K 99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99 E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N 281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N2 00D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281 Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N 200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E、E59K / K99E / N103Q / N200Q / C252S / K264E、E59K / K99E / N103Q / N281Q / C252S / K264E、E59K / K99E / N113Q / N200Q / C252S / K264E、E59K / K99E / N113Q / N281Q / C252S / K264E、E59K / K、 A heterodimer Fc fusion protein according to claim 1 or 2, having amino acid substitutions selected from the group consisting of 99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

4. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E) ), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), iii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59 Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) SEQ ID NO: 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) SEQ ID NO: 206 (IL-12p40 (E32Q / E59Q) )), xli) Sequence ID 207 (IL-12p40 (D34N / E59Q)), xliii) Sequence ID 208 (IL-12p40 (E59Q / E187Q)), xliiii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163 E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviiii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K 99E)), li) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E, 45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)) lxii) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N 103D / N113D), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Distribution Column number XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence number XXX (IL-12p40 (N103D / N200D / N281D)), lxxvi) Sequence number XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence number XXX (IL-12p40 (N103Q)), lxxviiii) Sequence number XXX (IL-12p40 (N113Q)), lxxix) Sequence number XXX (IL-12p40 (N200Q)), lxxx) Sequence number XXX (IL-12p40 (N281Q)),lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N113Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)), lxxxiiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40(N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX(IL-12p40(N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX(IL-12p40(N103Q / N200Q / N281Q)), xc) Sequence ID XXX(IL-12p40(N113Q / N200Q / N281Q)), xci) Sequence ID XXX(IL-12p40(N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX(IL-12p40(D34N / E59K / K99E )), xciii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E 59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)),ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N281Q / , C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N1 A heterodimer Fc fusion protein according to any one of claims 1 to 3, having a polypeptide sequence selected from the group consisting of 03Q / N113Q / N200Q / C252S / K264E)), cviii) SEQ ID NO: XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) SEQ ID NO: XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

5. The heterodimer Fc fusion protein according to any one of claims 1 to 4, wherein the variant IL-12p40 subunit has at least 90% identity with a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

6. The heterodimer Fc fusion protein according to any one of claims 1 to 4, wherein the IL-12p35 subunit is a variant IL-12p35 subunit.

7. The heterodimer Fc fusion protein according to any one of claims 1 to 4 and 6, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or the IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or the IL-12 receptor complex.

8. The heterodimer Fc fusion protein according to any one of claims 6 to 7, wherein the variant IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

9. The heterodimer Fc fusion protein according to any one of claims 6 to 8, wherein the variant IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

10. The variant IL-12p35 subunit is i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Sequence ID 118 (IL-12p35 (E38Q)), vii) Sequence ID 119 (IL-12p35 (N151D)), viiii) Sequence ID 120 (IL-12 p35 (Q135E), ix) Sequence ID 121 (IL-12p35 (Q35D)), x) Sequence ID 122 (IL-12p35 (Q146E)), xi) Sequence ID 123 (IL-12p35 (N76D)), xi) Sequence ID 124 (IL-12p35 (E162Q)), xiiii) Sequence ID 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) Sequence ID XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) I L-12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xx vii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)),xxxiv) Sequence ID XXX (IL-12p35 (M97A)), xxxv) Sequence ID XXX (IL-12p35 (L89A)), xxxvi) Sequence ID XXX (IL-12p35 (L124A)), xxxvii) Sequence ID XXX (IL-12p35 (M125A)), xxxviiii) Sequence ID XXX (IL-12p35 (L75A)), xxxiv) Sequence ID XXX (IL-12p35 (I171A)), xxxv) Sequence ID XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Column number XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence number XXX (IL-12p35 (N85Q / N195Q), xli) Sequence number XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence number XXX (IL-12p35 (N71D / N85D)), xliiii) Sequence number XXX (IL-12p35 (N71 A heterodimer Fc fusion protein according to any one of claims 6 to 9, having a polypeptide sequence selected from the group consisting of D / N195D), xlv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

11. The heterodimerized Fc fusion protein according to any one of claims 1 to 10, wherein the modification that promotes heterodimerization of the first and second Fc domains is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q; T366S / L368A / Y407V and T366W; T366S / L368A / Y407V / Y349C and T366W / S354C.

12. The heterodimer Fc fusion protein according to any one of claims 1 to 11, wherein the variant IL-12p40 subunit domain is bound to the first Fc domain using a first domain linker, and / or the IL-12p35 subunit domain is bound to the second Fc domain using a second domain linker.

13. The heterodimer Fc fusion protein according to any one of claims 1 to 12, wherein the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

14. The heterodimer Fc fusion protein according to any one of claims 1 to 13, wherein the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

15. The heterodimer Fc fusion protein according to any one of claims 1 to 14, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

16. The aforementioned heterodimer Fc fusion protein, a) i) Sequence ID 47 (XENP27201 Chain 1), ii) Sequence ID 85 (XenD24752), iii) Sequence ID 86 (XenD24753), iv) Sequence ID 87 (XenD24754), v) Sequence ID 88 (XenD24755), vi) Sequence ID 89 (XenD24756), vii) Sequence ID 90 (XenD24757), viiii) Sequence ID 91 (XenD24758), ix) Sequence ID 92 (XenD24759), x) Sequence ID 93 (XenD24760), xi) Sequence ID 94 (XenD24761), xii) Sequence ID 95 (XenD24762), xiii) Sequence ID 96 (XenD24763), xiv) Sequence ID 97 (XenD24764), xv) Sequence ID 98 (XenD24765), xvi) Sequence ID 99 (XenD24766), xvii) Sequence ID 100 (XenD24767), xviiii) Sequence ID 101 (XenD24768), xix) Sequence ID 102 (XenD24769), xx) Sequence ID 103 (XenD24770), xxi) Sequence ID 104 (XenD24771), xxii) Sequence ID 105 (XenD24772), x xiiii) Sequence ID 106 (XenD24773), xxiv) Sequence ID 107 (XenD24774), xxv) Sequence ID 108 (XenD24775), xxvi) Sequence ID 109 (XenD24776), xxvii) Sequence ID 110 (XenD24777), xxviiii) Sequence ID 111 (XenD24778), xxix) Sequence ID 112 (XenD24792), xxx) Sequence ID 215 (XenD25922), xxxi) Sequence ID 216 (XenD25923), xxxii) Sequence ID 217 (XenD25924), xx xiii) Sequence ID 218 (XenD25925), xxxiv) Sequence ID 219 (XenD25926), xxxv) Sequence ID 220 (XenD25927), xxxvi) Sequence ID 221 (XenD25928), xxxvii) Sequence ID 222 (XenD25929), xxxviiii) Sequence ID 223 (XenD25930), xxxix) Sequence ID 224 (XenD25931), xl) Sequence ID XXX (XenD26411), xli) Sequence ID XXX (XenD26412), xlii) Sequence ID XXX (XenD26413),xliiii) Sequence ID XXX (XenD26414), xliv) Sequence ID XXX (XenD26415), xlv) Sequence ID XXX (XenD26416), xlvi) Sequence ID XXX (XenD26417), xlvii) Sequence ID XXX (XenD26418), xlviiii) Sequence ID XXX (XenD2, 7070), xlix) Sequence ID XXX (XenD27071), l) Sequence ID XXX (XenD27072), li) Sequence ID XXX (XenD27073), lii) Sequence ID XXX (XenD27074), liiii) Sequence ID XXX (XenD27075), liv) Sequence ID XXX (XenD27076), lv) Sequence ID XXX (XenD27077), lvi) Sequence ID XXX (XenD27078), lvii) Sequence ID XXX (XenD28173), lviiii) Sequence ID XXX (XenD24876), lix) Sequence number XXX (XenD27162), lx) SEQ ID NO: XXX (XenD27163), lxi) SEQ ID NO: XXX (XenD27164), lxi) SEQ ID NO: XXX (XenD27165), lxi) SEQ ID NO: XXX (XENP31582, chain 1), lxi) SEQ ID NO: XXX (XENP31583, chain 1), lxv) SEQ ID NO: XXX (XENP31584, chain 1), lxvi) SEQ ID NO: XXX (XENP32187, chain 1), lxvii) SEQ ID NO: XXX (XENP32188, chain 1), lxviiii) SEQ ID NO: XXX (XENP32189, chain 1), lxix) SEQ ID NO: XXX (XENP32190 chain 1), lxx) SEQ ID NO: XXX (XENP32191 chain 1), lxxi) SEQ ID NO: XXX (XENP32991 chain 1), lxxii) SEQ ID NO: XXX (XENP32992 chain 1), lxxiii) SEQ ID NO: XXX (XENP32993 chain 1), lxxiv) SEQ ID NO: XXX (XENP32994 chain 1), lxxv) SEQ ID NO: XXX (XENP32995 chain 1), lxxvi) SEQ ID NO: XXX (XENP32996 chain 1), lxxvi) SEQ ID NO: XXX (XENP32997 chain 1), lxxvi i) SEQ ID NO: XXX (XENP32998 chain 1), lxxvii) SEQ ID NO: XXX (XENP32999 chain 1), lxxviiii) SEQ ID NO: XXX (XENP33000 chain 1), lxxix) SEQ ID NO: XXX (XENP33001 chain 1), lxxx) SEQ ID NO: XXX (XENP33002 chain 1), lxxxi) SEQ ID NO: XXX (XENP33003 chain 1), lxxxii) SEQ ID NO: XXX (XENP33004 chain 1), lxxxiiii) SEQ ID NO: XXX (XENP33005 chain 1), lxxxiv) SEQ ID NO: XXX (XENP33006 chain 1),The first fusion protein having a polypeptide sequence selected from the group consisting of lxxxv) SEQ ID NO: XXX (XENP33007 chain 1), lxxxvi) SEQ ID NO: XXX (XENP33008 chain 1), lxxxvii) SEQ ID NO: XXX (XENP33008 chain 1), lxxxviiii) SEQ ID NO: XXX (XENP33009 chain 1), lxxxix) SEQ ID NO: XXX (XENP33010 chain 1), and xc) SEQ ID NO: XXX (XENP33011 chain 1), b) i) SEQ ID NO: 48 (XENP27201 chain 2), ii) SEQ ID NO: 126 (XenD24779), iii) SEQ ID NO: 127 (XenD24780), iv) SEQ ID NO: 128 (XenD24781), v) SEQ ID NO: 129 (XenD24782), vi) SEQ ID NO: 130 (XenD24783), vii) SEQ ID NO: 131 (XenD24784), viiii) SEQ ID NO: 132 (XenD24785), ix) SEQ ID NO: 133 (XenD247 86), x) Sequence ID 134 (XenD24787), xi) Sequence ID 135 (XenD24788), xii) Sequence ID 136 (XenD24789), xiiii) Sequence ID 137 (XenD24790), xiv) Sequence ID 138 (XenD24791), xv) Sequence ID 236 (XenD25911), xvi) Sequence ID 237 (XenD25912), xvii) Sequence ID 238 (XenD25913), xviiii) Sequence ID 239 ( XenD25914), xix) Sequence ID 240 (XenD25915), xx) Sequence ID 241 (XenD25916), xxi) Sequence ID 242 (XenD25917), xxii) Sequence ID 243 (XenD25918), xxiii) Sequence ID 244 (XenD25919), xxiv) Sequence ID 245 (XenD25920), xxv) Sequence ID 246 (XenD25921), xxvi) Sequence ID XXX (XenD26427), xxvii) Sequence ID XXX (XenD26428), xxviiii) Sequence ID XXX (XenD27089), xxix) Sequence ID XXX (XenD27090), xxx) Sequence ID XXX (XenD27091), xxxi) Sequence ID XXX (XenD27092), xxxii) Sequence ID XXX (XenD27093), xxxiiii) Sequence ID XXX (XenD27094), xxxix) Sequence ID XXX (XenD24877), xl ) Sequence ID XXX (XenD27166), xli) Sequence ID XXX (XENP31582, chain 2), xlii) Sequence ID XXX (XENP31583, chain 2), xliiii) Sequence ID XXX (XENP31584, chain 2), xliv) Sequence ID XXX (XENP32187, chain 2), xlv) Sequence ID XXX (XENP32188, chain 2), xlvi) Sequence ID XXX (XENP32189, chain 2), xlvii) Sequence ID XXX (XENP32190, chain 2), xlviii) Sequence ID XXX (XENP32191 chain 2), xlix) Sequence ID XXX (XENP32991 chain 2), l) Sequence ID XXX (XENP32992 chain 2), li) Sequence ID XXX (XENP32993 chain 2), lii) Sequence ID XXX (XENP32994 chain 2), liiii) Sequence ID XXX (XENP32995 chain 2), liv) Sequence ID XXX (XENP32996 chain 2), lv) Sequence ID XXX (XENP32997 chain 2), lvi) Sequence ID XXX (XENP32998 chain 2), lvii) Column number XXX (XENP32999 chain 2), lviiii) Sequence number XXX (XENP33000 chain 2), lix) Sequence number XXX (XENP33001 chain 2), lx) Sequence number XXX (XENP33002 chain 2), lxi) Sequence number XXX (XENP33003 chain 2), lxi) Sequence number XXX (XENP33004 chain 2), lxiiii) Sequence number XXX (XENP33005 chain 2), lxiv) Sequence number XXX (XENP33006 chain 2), lxv) Sequence number XXX (XENP3 A heterodimer Fc fusion protein according to any one of claims 1 to 15, comprising the second fusion protein having a polypeptide sequence selected from the group consisting of 3007-chain 2), lxvi) SEQ ID NO: XXX (XENP33008-chain 2), lxvii) SEQ ID NO: XXX (XENP33008-chain 2), lxviiii) SEQ ID NO: XXX (XENP33009-chain 2), lxix) SEQ ID NO: XXX (XENP33010-chain 2), and lxx) SEQ ID NO: XXX (XENP33011-chain 2).

17. The aforementioned heterodimer Fc fusion proteins are XENP27201, XENP28820, XENP28821, XENP28822, XENP28823, XENP28824, XENP28825, XENP28826, XENP28827, XENP28828, XENP28829, XENP28830, XENP28831, XENP28832, XENP28833, XENP28834, XENP28835, XENP28836, XENP28837, XENP28838, XENP28839, XENP28840, XENP28841, XENP28842, XENP28843, XENP28844, XENP28845, XENP28846, XENP28847, XE NP28848, XENP28849, XENP28850, XENP28851, XENP28852, XENP29949, XENP2 9950, XENP29951, XENP29952, XENP30597, XENP30598, XENP30599, XENP306 00, XENP30601, XENP30602, XENP30603, XENP30604, XENP30605, XENP30606, XENP30307, XENP30308, XENP30609, XENP31250, XENP31251, XENP31252, XE NP31253, XENP31254, XENP31255, XENP31256, XENP31257, XENP31258, XENP3 1259, XENP31260, XENP31261, XENP31262, XENP31263, XENP31264, XENP312 65, XENP31286, XENP31142, XENP31143, XENP31144, XENP31145, XENP31146, XENP31582, XENP31583, XENP31584, XENP32186, XENP32187, XENP32188, XE NP32189, XENP32190, XENP32191, XENP32991, XENP32992, XENP32993, XENP3 2994, XENP32995, XENP32996, XENP32997, XENP32998, XENP32999, XENP330 00, XENP33001, XENP33002, XENP33003, XENP33004, XENP33005, XENP33006,Claims 1 to 1, selected from the group consisting of XENP33007, XENP33008, XENP33008, XENP33009, XENP33010, and XENP33011, A heterodimer Fc fusion protein as described in any one of item 16.

18. The heterodimer Fc fusion according to any one of claims 1 to 17, wherein the heterodimer Fc fusion protein is selected from the group consisting of XENP31251, XENP31254, XENP31258, XENP32186, XENP32187, XENP32188, XENP32189, XENP32190, and XENP32191.

19. The heterodimer Fc fusion protein according to any one of claims 17 to 18, further comprising one or more modifications to the IL-12p40 subunit selected from the group consisting of N103Q, N113Q, N200Q, and N281Q.

20. A composition comprising a heterodimer Fc fusion protein according to any one of claims 1 to 19 for use in the treatment of cancer in a subject.

21. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 1 to 19.

22. A host cell comprising one or more nucleic acids as described in claim 21.

23. A method for producing a heterodimer Fc fusion protein, the method comprising culturing the host cell described in claim 22 under conditions in which the heterodimer Fc fusion protein is produced.

24. A method for purifying a heterodimer Fc fusion protein according to any one of claims 1 to 19, wherein the method is a) To provide a composition containing the heterodimer Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the heterodimer Fc fusion protein.

25. A heterodimer Fc fusion protein, a) A first fusion protein comprising a first protein domain and a first Fc domain, wherein the first protein domain is covalently bonded to the C-terminus of the first Fc domain, b) A second fusion protein comprising a second protein domain and a second Fc domain, wherein the second protein domain is covalently bonded to the C-terminus of the Fc domain, A heterodimerized Fc fusion protein wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain includes an IL-12p40 subunit, and the second protein domain includes an IL-12p35 subunit.

26. The heterodimerized Fc fusion protein according to claim 25, wherein the modification that promotes heterodimerization of the first and second Fc domains is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including a cross-linked disulfide, T366S / L368A / Y407V / Y349C:T366W / S354C).

27. The heterodimer Fc fusion protein according to claim 25 or 26, wherein the first protein domain is bound to the first Fc domain using a first domain linker, and / or the second protein domain is bound to the second Fc domain using a second domain linker.

28. The heterodimer Fc fusion protein according to any one of claims 25 to 27, wherein the first and / or second Fc domain has an additional set of amino acid substitutions, including Q295E / N384D / Q418E / N421D, according to EU numbering.

29. The heterodimer Fc fusion protein according to any one of claims 25 to 28, wherein the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

30. The heterodimer Fc fusion protein according to any one of claims 25 to 29, wherein the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

31. The heterodimer Fc fusion protein according to any one of claims 25 to 30, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

32. The heterodimer Fc fusion protein according to any one of claims 25 to 31, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

33. The heterodimer Fc fusion protein according to any one of claims 25 to 32, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or the IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or the IL-12 receptor complex.

34. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P 185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S24 5. A heterodimer Fc fusion protein according to any one of claims 25 to 33, having one or more amino acid modifications with amino acid residues selected from the group consisting of Y246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

35. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K1 A heterodimer Fc fusion protein according to any one of claims 25 to 34, having one or more amino acid substitutions selected from the group consisting of 63E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

36. The IL-12p40 subunit is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E5 9Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K 99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99 E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N 281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N2 00D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281 Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N 200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E、E59K / K99E / N103Q / N200Q / C252S / K264E、E59K / K99E / N103Q / N281Q / C252S / K264E、E59K / K99E / N113Q / N200Q / C252S / K264E、E59K / K99E / N113Q / N281Q / C252S / K264E、E59K / K99E / N200Q / N281Q / C252S / K264E、E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E、E59K / K99E / N1、 A heterodimer Fc fusion protein according to any one of claims 25 to 35, having amino acid substitutions selected from the group consisting of 03Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

37. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E) ), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), iii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59 Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) SEQ ID NO: 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) SEQ ID NO: 206 (IL-12p40 (E32Q / E59Q)), xli) SEQ ID NO: 207 (IL-12 p40 (D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40 (E59Q / E187Q)), xliiii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xl vii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviiii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), li) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12 p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59, K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103 D / N281D), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)) ), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)), lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)),lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N28 1Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E 32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N c) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N281Q / C252S / K264E)),civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XX, X (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID X A heterodimer Fc fusion protein according to any one of claims 25 to 36, having a polypeptide sequence selected from the group consisting of XX (IL-12p40 (E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

38. The IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N1 A heterodimer Fc fusion protein according to any one of claims 25 to 37, having one or more amino acid modifications as amino acid residues selected from the group consisting of 51, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

39. The heterodimer Fc fusion protein according to any one of claims 25 to 38, wherein the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

40. The heterodimer Fc fusion protein according to any one of claims 25 to 39, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

40. The IL-12p35 subunits are: i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Sequence ID 118 (IL-12p35 (E38Q)), vii) Sequence ID 119 (IL-12p35 (N151D)), viiii) Sequence ID 120 (IL-12p35 (Q135E)), ix) Sequence ID 121 (IL-12p35 (Q35D)), x ) Sequence ID 122 (IL-12p35 (Q146E)), xi) Sequence ID 123 (IL-12p35 (N76D)), xi) Sequence ID 124 (IL-12p35 (E162Q)), xiiii) Sequence ID 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) Sequence ID XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL-12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 ( IL-12p35(E153K)), xxii) IL-12p35(K158E), xxiii) IL-12p35(D165N), xxiv) Sequence ID 225 (IL-12p35(N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35(N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35(Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35(N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35(E79Q / N151D)), xxx) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)), xxxiv) Sequence ID XXX (IL-12p35 (M97A)), xxxv) Sequence ID XXX (IL-12p35 (L89A)), xxxvi) Sequence ID XXX (IL-12p35 (L124A)), xxxvii) Sequence ID XXX (IL-12p35 (M125A)), xxxviiii) Sequence ID XXX (IL-12p35 (L75A)), xxxiv) Sequence ID XXX (IL-12p35 (I171A)), xxxv) Sequence ID XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q )), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Sequence ID XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)), xliiii) Sequence ID XXX (IL-12p35 (N71D / N195D), xliv) Sequence ID XXX (IL-12p35 (N85D / N195D)), xlv) Sequence ID XXX (IL-12p35 (D55Q)),A heterodimer Fc fusion protein according to any one of claims 25 to 40, having a polypeptide sequence selected from the group consisting of and xlvi) SEQ ID NO XXX (IL-12p35 (D55K)).

41. The aforementioned heterodimer Fc fusion protein, a) The first fusion protein having the polypeptide sequence of Sequence ID No. 49 (XENP27202 chain 1), b) The heterodimer Fc fusion protein according to any one of claims 25 to 40, comprising the second fusion protein having the polypeptide sequence of Sequence ID No. 50 (XENP27202 chain 2).

42. The heterodimer Fc fusion protein according to any one of claims 25 to 41, wherein the heterodimer Fc fusion protein is XENP27202.

43. A composition comprising a heterodimer Fc fusion protein according to any one of claims 25 to 42 for use in the treatment of cancer in a subject.

44. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 25 to 42.

45. A host cell comprising one or more nucleic acids as described in claim 44.

46. A method for producing a heterodimer Fc fusion protein, the method comprising culturing the host cell described in claim 45 under conditions that produce the heterodimer Fc fusion protein.

47. A method for purifying a heterodimer Fc fusion protein according to any one of claims 25 to 42, wherein the method is a) To provide a composition containing the heterodimer Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the heterodimer Fc fusion protein.

48. A heterodimer Fc fusion protein, a) A fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is covalently bonded to the second protein domain, and the second protein domain is covalently bonded to the N-terminus of the first Fc domain, b) Including a second Fc domain, A heterodimerized Fc fusion protein wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain includes an IL-12p40 subunit, and the second protein domain includes an IL-12p35 subunit.

49. The heterodimerized Fc fusion protein according to claim 48, wherein the modification that promotes heterodimerization of the first and second Fc domains is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including a cross-linked disulfide, T366S / L368A / Y407V / Y349C:T366W / S354C).

50. The heterodimer Fc fusion protein according to claim 48 or 49, wherein the first protein domain is bound to the second protein domain using a first domain linker, and / or the second protein domain is bound to the first Fc domain using a second domain linker.

51. The heterodimer Fc fusion protein according to any one of claims 48 to 50, wherein the first and / or second Fc domain has an additional set of amino acid substitutions comprising Q295E / N384D / Q418E / N421D according to EU numbering.

52. The heterodimer Fc fusion protein according to any one of claims 48 to 51, wherein the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

53. The IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence). A heterodimer Fc fusion protein according to any one of claims 48 to 52, having a polypeptide sequence.

54. The heterodimer Fc fusion protein according to any one of claims 48 to 53, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

55. The heterodimer Fc fusion protein according to any one of claims 48 to 53, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

56. The heterodimer Fc fusion protein according to any one of claims 48 to 55, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

57. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y 66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E11 0, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174 A heterodimer Fc fusion protein according to any one of claims 48 to 56, having one or more amino acid modifications with amino acid residues selected from the group consisting of A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, Q256, K158, C252, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

58. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K1 A heterodimer Fc fusion protein according to any one of claims 48 to 57, having one or more amino acid substitutions selected from the group consisting of 63E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

59. The IL-12p40 subunit is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K9 9E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K - 44E、EK9K / E99E / Q14KEEK9K / K99E / R159E、E59K / K99E / 265E、D18K / ED9E / K99E / K264E、DI8K / E59K / K99E / C252S、D18K / E59K / K99E / C252E / G251E、ED 9K / K99Y / C252S、E59K / K99E / C252S / K264E、EK9K / K99E / C252S、N103D / 1113 D、1103D / 200D、1100 / 200D、1110 / 2000、1110 / 2000、1000 / / 2000、 N103D / N113D / N200D、N103 / / N113D / N281D、N103 / / N200 / / 200D、N113D / 2 00D / N281D、103Q / N113Q、103Q / .200Q、103Q / .281Q、1113Q / .200Q、11 3Q / N281Q、N200Q / N281Q、103Q / / 1111 / 2001、1101 / / 11111 / .2811、1111 / N200Q / N281Q、N113Q / / 200Q / N201Q、N103Q / N113Q / N2001 / 720119E99999 9E / N103Q / C252S / K26EEEE9K / K99E / N113Q / C252S / K264E、ED9K / .99E / N2E 0Q / C252S / 526EEE59K / E99E / N281Q / / C252S / 5261E、E59K / K99E / N103Q / 1 13Q / C252S / K261E、E59K / 999EN103Q / N200Q / C252S / K261E、E19K / 999E / 11 03Q / N281Q / C252S / 5261E、E19K / 999E / N113Q / N200Q / C252S / 5251E 99E / N113Q / N281Q / C252S / K26EE、E9E / / 99E / N200Q / N281Q / / ND252.A heterodimer Fc fusion protein according to any one of claims 48 to 58, having amino acid substitutions selected from the group consisting of E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

60. The IL-12p40 subunit is i) SEQ ID NO: 57 (IL-12p40 (N103D)), ii) SEQ ID NO: 58 (IL-12p40 (N113D)), iii) SEQ ID NO: 59 (IL-12p40 (N200D)), iv) SEQ ID NO: 60 (IL-12p40 (N281D)), v) SEQ ID NO: 61 (IL-12p40 (N103D / N113D / N200D / N281D)), vi) SEQ ID NO: 62 (IL-12p4 0 (Q42E)), vii) SEQ ID NO: 63 (IL-12p40 (E45Q)), viiii) SEQ ID NO: 64 (IL-12p40 (Q56E)), ix) SEQ ID NO: 65 (IL-12p40 (E59Q)), x) SEQ ID NO: 66 (IL-12p40 (D62N)), xi) SEQ ID NO: 67 (IL-12p40 (Q42E / E45Q)), xi) SEQ ID NO: 68 (IL-12p40 (E45Q) / Q56E)), xiii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40 (D16 1N)), xviiii) SEQ ID NO: 74 (IL-12p40(E73Q)), xix) SEQ ID NO: 75 (IL-12p40(Q144E)), xx) SEQ ID NO: 76 (IL-12p40(E262Q)), xxi) SEQ ID NO: 77 (IL-12p40(E100Q)), xxi) SEQ ID NO: 78 (IL-12p40(D18N)), xiii) SEQ ID NO: 79 (IL-12p40(E33 Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q)), xxvi) SEQ ID NO: 82 (IL-12p40 (Q256N)), xxvii) SEQ ID NO: 83 (IL-12p40 (E299Q)), xxviiii) SEQ ID NO: 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) SEQ ID NO: XXX (IL-1 2p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) Sequence ID 206 (IL-12p40 (E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40 (D34 N / E59Q)), xlii) Sequence ID 208 (IL-12p40 (E59Q / E187Q)), xliiii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviiii) Sequence ID 214 (IL-12p40 (E59Q / K26 0E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), li) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)),lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxii) Sequence ID XX X (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N11 3D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID X XX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)),lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N11, 3Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xc iv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E c) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)),ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113 A heterodimer Fc fusion protein according to any one of claims 48 to 59, having a polypeptide sequence selected from the group consisting of Q / N200Q / C252S / K264E), cviii) SEQ ID NO: XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) SEQ ID NO: XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

61. The IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N1 A heterodimer Fc fusion protein according to any one of claims 48 to 60, having one or more amino acid modifications as amino acid residues selected from the group consisting of 51, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

62. The IL-12p35 subunits are N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N1 A heterodimer Fc fusion protein according to any one of claims 48 to 61, having one or more amino acid substitutions selected from the group consisting of 36D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

63. The heterodimer Fc fusion protein according to any one of claims 48 to 62, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

64. The aforementioned IL-12p35 subunit is i) SEQ ID NO: 113 (IL-12p35 (N71D)), ii) SEQ ID NO: 114 (IL-12p35 (N85D)), iii) SEQ ID NO: 115 (IL-12p35 (N195D)), iv) SEQ ID NO: 116 (IL-12p35 (N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35 (E153Q)), vi) SEQ ID NO: 118 (IL-12p35 (E38Q)), vii) SEQ ID NO: 119 (IL-12p35 (N151D)), viiii) SEQ ID NO: 120 (IL-12p35 (Q135E)), ix) SEQ ID NO: 121 (IL -12p35 (Q35D)), x) SEQ ID NO: 122 (IL-12p35 (Q146E)), xi) SEQ ID NO: 123 (IL-12p35 (N76D)), xii) SEQ ID NO: 124 (IL-12p35 (E162Q)), xiiii) SEQ ID NO: 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) SEQ ID NO: XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL-12p35 (N136D), xix) IL-12p3 5 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35 (N151D / K 158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)), xxxiv) Sequence ID XXX (IL-12p35 (M97A)),xxxv) SEQ ID NO: XXX (IL-12p35 (L89A)), xxxvi) SEQ ID NO: XXX (IL-12p35 (L124A)), xxxvii) SEQ ID NO: XXX (IL-12p35 (M125A)), xxxviiii) SEQ ID NO: XXX (IL-12p35 (L75A)), xxxiv) SEQ ID NO: XXX (IL-12p35 (I171A)), xxxv) SEQ ID NO: XXX (IL-12p35 (N71Q)), xxxvi) SEQ ID NO: XXX (IL-12p35 (N85Q)), xxxvii) SEQ ID NO: XX X (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Sequence ID XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)), xliiii) Sequence ID XXX (IL-12p35 (N71D / N1, A heterodimer Fc fusion protein according to any one of claims 48 to 63, having a polypeptide sequence selected from the group consisting of 95D), xlv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

65. The aforementioned heterodimer Fc fusion protein, a) The fusion protein having the polypeptide sequence of Sequence ID No. 51 (XENP27203 chain 1), b) The heterodimer Fc fusion protein according to any one of claims 48 to 64, comprising the second Fc domain having the polypeptide sequence of Sequence ID No. 52 (XENP27203 chain 2).

66. The heterodimer Fc fusion protein according to any one of claims 48 to 65, wherein the heterodimer Fc fusion protein is XENP27203.

67. The heterodimer Fc fusion protein according to any one of claims 48 to 65, wherein the heterodimer Fc fusion protein is XENP31290.

68. A composition comprising a heterodimer Fc fusion protein according to any one of claims 48 to 66 for use in the treatment of cancer in a subject.

69. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 48 to 67.

70. A host cell comprising one or more nucleic acids as described in claim 69.

71. A method for producing a heterodimer Fc fusion protein, the method comprising culturing the host cell described in claim 70 under conditions that produce the heterodimer Fc fusion protein.

72. A method for purifying a heterodimer Fc fusion protein according to any one of claims 48 to 67, wherein the method is a) To provide a composition containing the heterodimer Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the heterodimer Fc fusion protein.

73. A heterodimer Fc fusion protein, a) A fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is covalently bonded to the second protein domain, and the second protein domain is covalently bonded to the N-terminus of the first Fc domain, b) Including a second Fc domain, A heterodimerized Fc fusion protein wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain includes an IL-12p35 subunit, and the second protein domain includes an IL-12p40 subunit.

74. The modifications that promote heterodimerization of the first and second Fc domains are, according to EU numbering, L368D / K370S and S364K;L368D / K370S and The heterodimer Fc fusion protein according to claim 73, which is a set of amino acid substitutions selected from the group consisting of S364K / E357L;L368D / K370S and S364K / E357Q;T411E / K360E / Q362E and D401K;L368E / K370S and S364K;K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including a cross-linked disulfide, T366S / L368A / Y407V / Y349C:T366W / S354C).

75. The heterodimer Fc fusion protein according to claim 73 or 74, wherein the first protein domain is bound to the second protein domain using a first domain linker, and / or the second protein domain is bound to the first Fc domain using a second domain linker.

76. The heterodimer Fc fusion protein according to any one of claims 73 to 75, wherein the first and / or second Fc domain has an additional set of amino acid substitutions comprising Q295E / N384D / Q418E / N421D according to EU numbering.

77. The heterodimer Fc fusion protein according to any one of claims 73 to 76, wherein the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

78. The heterodimer Fc fusion protein according to any one of claims 73 to 77, wherein the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and / or SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

79. The heterodimer Fc fusion protein according to any one of claims 73 to 78, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

80. The heterodimer Fc fusion protein according to any one of claims 73 to 79, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

81. The heterodimer Fc fusion protein according to any one of claims 73 to 80, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

82. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y 66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E11 0, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174 A heterodimer Fc fusion protein according to any one of claims 73 to 81, having one or more amino acid modifications with amino acid residues selected from the group consisting of A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

83. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K1 A heterodimer Fc fusion protein according to any one of claims 73 to 82, having one or more amino acid substitutions selected from the group consisting of 63E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

84. The IL-12p40 subunits are N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K 163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E5 9K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N1 03D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N200D / N281D, N 113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200 Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59 K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N 200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N11 A heterodimer Fc fusion protein according to any one of claims 73 to 83, having amino acid substitutions selected from the group consisting of 3Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

85. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E) ), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), iii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59 Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) SEQ ID NO: 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) SEQ ID NO: 206 (IL-12p40 (E32Q / E59Q)), xli) SEQ ID NO: 207 (IL-12p40 (D34N / E59Q)), xliii) SEQ ID NO: 208 (IL-12p40 (E59Q / E187Q)), xliiii) SEQ ID NO: 209 (IL-12 p40 (S43E / E59Q)), xlv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), l) Distribution, Column number XXX (IL-12p40 (D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E) 9E / Q144K), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99 E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)),lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)), lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), l xxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59 K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)),c) Sequence ID XXX (IL-12p40(E59K / K99E / N281Q / C252S / K264E, )), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N 103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX ( IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N A heterodimer Fc fusion protein according to any one of claims 73 to 84, having a polypeptide sequence selected from the group consisting of (200Q / N281Q / C252S / K264E)) and (cix) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

86. The IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N1 A heterodimer Fc fusion protein according to any one of claims 73 to 85, having one or more amino acid modifications as amino acid residues selected from the group consisting of 51, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

87. The heterodimer Fc fusion protein according to any one of claims 73 to 86, wherein the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

88. The heterodimer Fc fusion protein according to any one of claims 73 to 87, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

89. The IL-12p35 subunits are: i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E1 53Q)), vi) Sequence ID 118 (IL-12p35(E38Q)), vii) Sequence ID 119 (IL-12p35(N151D)), viiii) Sequence ID 120 (IL-12p35(Q135E)), ix) Sequence ID 121 (IL-12p35(Q35D)), x) Sequence ID 122 (IL-12p35(Q146E)), xi) Sequence ID 123 (IL- 12p35 (N76D)), xii) Sequence ID 124 (IL-12p35 (E162Q)), xiii) Sequence ID 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) Sequence ID XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL-12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (I L-12p35 (E153K), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xx ix) SEQ ID NO: 232 (IL-12p35 (D55Q / N151D)), xxx) SEQ ID NO: 233 (IL-12p35 (N136D / N151D)), xxxi) SEQ ID NO: 234 (IL-12p35 (N21D / N151D)), xxxii) SEQ ID NO: 235 (IL-12p35 (E143Q / N151D)), xxxiii) SEQ ID NO: XXX (IL-12p35 (F96A)), xxxiv) SEQ ID NO: XXX (IL-12p35 (M97A)), xxxv) SEQ ID NO: XXX (IL-12p35 (L89A)), xxxvi) SEQ ID NO: XX X (IL-12p35 (L124A)), xxxvii) Sequence ID XXX (IL-12p35 (M125A)), xxxviiii) Sequence ID XXX (IL-12p35 (L75A)), xxxiv) Sequence ID XXX (IL-12p35 (I171A)), xxxv) Sequence ID XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)),xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q)), xli) Sequence ID XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)), xliiii) Sequence ID XXX (IL-12p35 (N A heterodimer Fc fusion protein according to any one of claims 73 to 88, having a polypeptide sequence selected from the group consisting of 71D / N195D), xlv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

90. The aforementioned heterodimer Fc fusion protein, a) The fusion protein having the polypeptide sequence of Sequence ID No. 53 (XENP27204 chain 1), b) The heterodimer Fc fusion protein according to any one of claims 73 to 89, comprising the second Fc domain having the polypeptide sequence of Sequence ID No. 54 (XENP27204 chain 2).

91. The heterodimer Fc fusion protein according to any one of claims 73 to 90, wherein the heterodimer Fc fusion protein is XENP27204.

92. A composition comprising a heterodimer Fc fusion protein according to any one of claims 73 to 91 for use in the treatment of cancer in a subject.

93. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 73 to 91.

94. A host cell comprising one or more nucleic acids as described in claim 93.

95. A method for producing a heterodimer Fc fusion protein, the method comprising culturing the host cell described in claim 94 under conditions that produce the heterodimer Fc fusion protein.

96. A method for purifying a heterodimer Fc fusion protein according to any one of claims 73 to 91, wherein the method is a) To provide a composition containing the heterodimer Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the heterodimer Fc fusion protein.

97. A heterodimer Fc fusion protein, a) A fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is bound to the C-terminus of the first Fc domain, and the second protein domain is covalently bound to the first protein domain, b) Including a second Fc domain, A heterodimerized Fc fusion protein wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain includes an IL-12p40 subunit, and the second protein domain includes an IL-12p35 subunit.

98. The heterodimerized Fc fusion protein according to claim 97, wherein the modification that promotes heterodimerization of the first and second Fc domains is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including a cross-linked disulfide, T366S / L368A / Y407V / Y349C:T366W / S354C).

99. The heterodimer Fc fusion protein according to claim 97 or 98, wherein the first protein domain is bound to the second protein domain using a first domain linker, and / or the second protein domain is bound to the first Fc domain using a second domain linker.

100. The heterodimer Fc fusion protein according to any one of claims 97 to 99, wherein the first and / or second Fc domain has an additional set of amino acid substitutions comprising Q295E / N384D / Q418E / N421D according to EU numbering.

101. The heterodimer Fc fusion protein according to any one of claims 97 to 100, wherein the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

102. The heterodimer Fc fusion protein according to any one of claims 97 to 101, wherein the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

103. The heterodimer Fc fusion protein according to one of claims 97 to 102, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

104. The heterodimer Fc fusion protein according to any one of claims 97 to 103, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

105. The heterodimer Fc fusion protein according to any one of claims 97 to 104, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

106. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y 66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E11 0, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174 A heterodimer Fc fusion protein according to any one of claims 97 to 105, having one or more amino acid modifications with amino acid residues selected from the group consisting of A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

107. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K16 A heterodimer Fc fusion protein according to any one of claims 97 to 106, having one or more amino acid substitutions selected from the group consisting of 3E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

108. The IL-12p40 subunit is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E5 9Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K 99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99 E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N 281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N2 00D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281 Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N 200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N A heterodimer Fc fusion protein according to any one of claims 97 to 107, having amino acid substitutions selected from the group consisting of 103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

109. The IL-12p40 subunit is: i) Sequence ID 57 (IL-12p40 (N103D)), ii) Sequence ID 58 (IL-12p40 (N113D)), iii) Sequence ID 59 (IL-12p40 (N200D)), iv) Sequence ID 60 (IL-12p40 (N281D)), v) Sequence ID 61 (IL-12p40 (N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40 (Q42E)), vii) Sequence ID 63 (IL-12p40 (E45Q)), viiii) Sequence ID 64 (IL-1 2p40 (Q56E)), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xi) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), xiiii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL -12p40 (E45Q / Q56E / E59Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) Sequence ID 81 (IL-12p40 (E235Q)), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N10 3Q / N113Q / N200Q / N281Q / C252S / K264E), xxxiv) Sequence ID 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) Sequence ID 206 (IL-12p40 (E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40 (D34N / E59Q)), xl ii) Sequence ID 208 (IL-12p40 (E59Q / E187Q)), xliii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xlv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvii) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40 (E59Q / K260E)),xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), li) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)), liii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K9 9E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi ) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)),lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N10, 3D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)), lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40 (N 103Q / N200Q), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N 281Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)),c) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)) , ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252 S / K264E)), cvi) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID XXX (IL-12p40 (E59K / K9 A heterodimer Fc fusion protein according to any one of claims 97 to 1081, having a polypeptide sequence selected from the group consisting of 9E / N103Q / N200Q / N281Q / C252S / K264E)) and cix) SEQ ID NO XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

110. The IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143 The present invention relates to any one of claims 97 to 109, having one or more amino acid modifications as amino acid residues selected from the group consisting of Q146, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196. Heterodimer Fc fusion protein.

111. The heterodimer Fc fusion protein according to any one of claims 97 to 110, wherein the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

112. The heterodimer Fc fusion protein according to any one of claims 97 to 111, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

113. The IL-12p35 subunit is i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Sequence ID 118 (IL-12p35 (E38Q)), vii) Sequence ID 119 (IL-12p35 (N151D)), viiii) Sequence ID 120 (IL-12p35 ( Q135E), ix) SEQ ID NO: 121 (IL-12p35 (Q35D)), x) SEQ ID NO: 122 (IL-12p35 (Q146E)), xi) SEQ ID NO: 123 (IL-12p35 (N76D)), xi) SEQ ID NO: 124 (IL-12p35 (E162Q)), xiiii) SEQ ID NO: 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) SEQ ID NO: XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL- 12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxv ii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)),xxxiv) SEQ ID NO: XXX (IL-12p35 (M97A)), xxxv) SEQ ID NO: XXX (IL-12p35 (L89A)), xxxvi) SEQ ID NO: XXX (IL-12p35 (L124A)), xxxvii) SEQ ID NO: XXX (IL-12p35 (M125A)), xxxviiii) SEQ ID NO: XXX (IL-12p35 (L75A)), xxxiv) SEQ ID NO: XXX (IL-12p35 (I171A)), xxxv) SEQ ID NO: XXX (IL-12p35 (N71Q)), xxxvi) SEQ ID NO: XXX (IL-12p35 (N85Q)), xxxvii) SEQ ID NO: XXX (IL-12p3, 5 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Sequence ID XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)), x A heterodimer Fc fusion protein according to any one of claims 97 to 112, having a polypeptide sequence selected from the group consisting of liiii) SEQ ID NO: XXX (IL-12p35 (N71D / N195D), xlv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

114. A composition comprising a heterodimer Fc fusion protein according to any one of claims 97 to 113 for use in the treatment of cancer in a subject.

115. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 97 to 113.

116. A host cell comprising one or more nucleic acids as described in claim 115.

117. A method for producing a heterodimer Fc fusion protein, the method comprising culturing the host cell described in claim 116 under conditions that produce the heterodimer Fc fusion protein.

118. A method for purifying a heterodimer Fc fusion protein according to any one of claims 97 to 113, wherein the method is a) To provide a composition containing the heterodimer Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the heterodimer Fc fusion protein.

119. A heterodimer Fc fusion protein, a) A fusion protein comprising a first protein domain, a second protein domain, and a first Fc domain, wherein the first protein domain is bound to the C-terminus of the first Fc domain, and the second protein domain is covalently bound to the first protein domain, b) Including a second Fc domain, A heterodimerized Fc fusion protein wherein the first and second Fc domains include modifications that promote heterodimerization of the first and second Fc domains, the first protein domain includes an IL-12p35 subunit, and the second protein domain includes an IL-12p40 subunit.

120. The heterodimerized Fc fusion protein according to claim 119, wherein the modification that promotes heterodimerization of the first and second Fc domains is a set of amino acid substitutions selected from the group consisting of L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q and T366S / L368A / Y407V:T366W (optionally including a cross-linked disulfide, T366S / L368A / Y407V / Y349C:T366W / S354C).

121. The heterodimer Fc fusion protein according to claim 119 or 120, wherein the first protein domain is bound to the second protein domain using a first domain linker, and / or the second protein domain is bound to the first Fc domain using a second domain linker.

122. The heterodimer Fc fusion protein according to any one of claims 119 to 121, wherein the first and / or second Fc domain has an additional set of amino acid substitutions comprising Q295E / N384D / Q418E / N421D according to EU numbering.

123. The heterodimer Fc fusion protein according to any one of claims 119 to 122, wherein the first and / or second Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

124. The heterodimer Fc fusion protein according to any one of claims 119 to 123, wherein the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

125. The heterodimer Fc fusion protein according to any one of claims 119 to 124, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

126. The heterodimer Fc fusion protein according to any one of claims 119 to 125, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

127. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, A heterodimer Fc fusion protein according to any one of claims 119 to 126, having one or more amino acid modifications with amino acid residues selected from the group consisting of S245, Y246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

128. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K16 A heterodimer Fc fusion protein according to any one of claims 119 to 127, having one or more amino acid substitutions selected from the group consisting of 3E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

129. The IL-12p40 subunit is N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E5 9Q / K163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K 99E, E59K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99 E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N103D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N 281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N2 00D / N281D, N113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281 Q, N113Q / N200Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N 200Q / N281Q, E59K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E,E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E5 9K / K99E / N113Q / N200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K The amino acid substitutions selected from the group consisting of / K99E / N103Q / N113Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E, according to any one of claims 119 to 128, Heterodimer Fc fusion protein.

130. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E) ), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), iii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59 Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) SEQ ID NO: 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) SEQ ID NO: 206 (IL-12p40 (E32Q / E59Q)), xli) SEQ ID NO: 207 (IL-12p40 (D34N / E59Q)), xlii) SEQ ID NO: 208 (IL-12p40 (E5 9Q / E187Q), xliiii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviiii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), l) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)), liii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K 99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C, 252S)), lxii) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX ( IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N2 00D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxvii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX ( IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40(N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40(N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40(N200Q / N281Q)),lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p4 0 (D34N / E59K / K99E)), xciii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence number Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)),cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / C252S / , A heterodimer Fc fusion protein according to any one of claims 119 to 129, having a polypeptide sequence selected from the group consisting of K264E)), cviii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

131. The IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N15 1. A heterodimer Fc fusion protein according to any one of claims 119 to 130, having one or more amino acid modifications as amino acid residues selected from the group consisting of E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

132. The heterodimer Fc fusion protein according to any one of claims 119 to 131, wherein the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

133. The heterodimer Fc fusion protein according to any one of claims 119 to 132, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

134. The IL-12p35 subunit is i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Distribution Column number 118 (IL-12p35 (E38Q)), vii) Sequence number 119 (IL-12p35 (N151D)), viiii) Sequence number 120 (IL-12p35 (Q135E)), ix) Sequence number 121 (IL-12p35 (Q35D)), x) Sequence number 122 (IL-12p35 (Q146E)), xi) Sequence number 123 (IL-12p35 (N76D)), xi) Sequence number Sequence ID 124 (IL-12p35 (E162Q)), xiii) Sequence ID 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) Sequence ID XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL-12p35 (N136D), xix) IL-12p 35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N15 1D / D165N), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID Number 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)), xxxiv) Sequence ID XXX (IL-12p35 (M97A)), xxxv) Sequence ID XXX (IL-12p35 (L89A)), xxxvi) Sequence ID XXX (IL-12p35 (L124A)), xxxvii) Sequence ID XXX (IL-12p35 (M125A)), xxxviiii) Sequence ID XXX (IL-12p35 (L75A)), xxxiv) Sequence ID XXX (IL-12p3 5 (I171A)), xxxv) Sequence ID XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Sequence ID XXX (IL-12p35 (N71Q / N85 A heterodimer Fc fusion protein according to any one of claims 119 to 133, having a polypeptide sequence selected from the group consisting of Q / N195Q), xlii)Sequence ID XXX (IL-12p35 (N71D / N85D)), xliiii)Sequence ID XXX (IL-12p35 (N71D / N195D), xliv)Sequence ID XXX (IL-12p35 (N85D / N195D)), xlv)Sequence ID XXX (IL-12p35 (D55Q)), and xlvi)Sequence ID XXX (IL-12p35 (D55K)).

135. A composition comprising a heterodimer Fc fusion protein according to any one of claims 119 to 134 for use in the treatment of cancer in a subject.

136. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 119 to 134.

137. A host cell comprising one or more nucleic acids as described in claim 136.

138. A method for producing a heterodimer Fc fusion protein, the method comprising culturing the host cell described in claim 137 under conditions that produce the heterodimer Fc fusion protein.

139. A method for purifying a heterodimer Fc fusion protein according to any one of claims 119 to 134, wherein the method is a) To provide a composition containing the heterodimer Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the heterodimer Fc fusion protein.

140. a) IL-12p40 subunit, b) A heterodimer complex containing the IL-12p35 subunit.

141. The heterodimer complex according to claim 140, wherein the IL-12p40 subunit is covalently bonded to the IL-12p35 subunit using a domain linker.

142. The heterodimer complex according to claim 140 or 141, wherein the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and / or the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

143. The heterodimer complex according to any one of claims 140 to 142, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

144. The heterodimer complex according to any one of claims 140 to 143, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

145. The heterodimer complex according to any one of claims 140 to 144, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

146. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E 110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D1 A heterodimer complex according to any one of claims 140 to 145, having one or more amino acid modifications with amino acid residues selected from the group consisting of 74, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, C252, Q256, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

147. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, A heterodimer complex according to any one of claims 140 to 146, having one or more amino acid substitutions selected from the group consisting of K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

148. The IL-12p40 subunit is N103D / N113D / N200D / N28 1D、Q42E / E45Q、E45Q / Q56E、Q42E / E59Q、Q56E / E59Q、Q42E / E45Q / Q56E、E45Q / Q56E / E59Q、E32Q / E59Q、D34N / E59K、D34N / E59K / K99E、D34K / E59K / K99E、E32Q / D34N / E59K / K99E、E32K / D34N / E59K / K99E、D34N / E59Q、E59Q / E187Q、S43E / E59Q、S43K / E49Q、E59Q / K163E、E59Q / K99E、E59Q / K258E、E59Q / K260E、E59K / K99E、D18K / E59K / K99E、E59K / K99E / K264E、E59K / K99Y、E59Y / K99Y、E59Y / K99E、E45K / E59K / K99E、E59K / K99E / Q144E、E59K / K99E / Q144K、E59K / K99E / R159E、E59K / K99E / K264E、D18K / E59K / K99E / K264E、DI8K / E59K / K99E / C252S、D18K / E59K / K99E / C252S / K264E、E59K / K99Y / C252S、E59K / K99E / C252S / K264E、E59K / K99E / C252S、N103D / N113D、N103D / N200D、N103D / N281D、N113D / N200D、N113D / N281D、N200D / N281D、N103D / N113D / N200D、N103D / N113D / N281D、N103D / N200D / N281D、N113D / N200D / N281D、N103Q / N113Q、N103Q / N200Q、N103Q / N281Q、N113Q / N200Q、N113Q / N281Q、N200Q / N281Q、N103Q / N113Q / N200Q、N103Q / N113Q / N281Q、N103Q / N200Q / N281Q、N113Q / N200Q / N281Q、N103Q / N113Q / N200Q / N281Q、E59K / K99E / N103Q / C252S / K264E、E59K / K99E / N113Q / C252S / K264E、E59K / K99E / N200Q / C252S / K264E、E59K / K99E / N281Q / C252S / K264E、E59K / K99E / N103Q / N113Q / C252S / K264E、E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K2 64E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200 A heterodimer complex according to any one of claims 140 to 147, having amino acid substitutions selected from the group consisting of Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

149. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E)), ix) Sequence ID 65 (IL-12p40(E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xi) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), xiiii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59Q)), xvii) Sequence ID 73 (IL-12p40 (D161N)), xviiii) Sequence ID 74 (IL-12p40 (E73Q) ), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q)), xxvi) SEQ ID NO: 82 (IL-12p40 (Q256N)), xxvii) SEQ ID NO: 83 (IL-12p4 0 (E299Q), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40(K163E), xxxvii)IL-12p40(E187Q), xxxviiii)IL-12p40(K258E), xxxix)IL-12p40(K260E), xl)Sequence ID 206(IL-12p40(E32Q / E59Q)), xli)Sequence ID 207(IL-12p40(D34N / E59Q)), xlii)Sequence ID 208(IL-12p40(E59Q / E187Q)), xliiii)Sequence ID 209(IL-12p40(S43E / E59Q)), xliv)Sequence ID 210(IL-12p40(S43K / E 49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvii) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), l) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)),lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E 59K / K99E / R159E)), lx) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 ( E59K / K99Y / C252S), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D) ), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)),lxxvii) Sequence ID XXX (IL-12, p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)), lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12 p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL -12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), x cvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)),ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N1 03Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q A heterodimer complex according to any one of claims 140 to 148, having a polypeptide sequence selected from the group consisting of ( / N200Q / N281Q / C252S / K264E)) and (cix) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

150. The IL-12p35 subunit comprises Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, A heterodimer complex according to any one of claims 140 to 149, having one or more amino acid modifications as amino acid residues selected from the group consisting of N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

151. The heterodimer complex according to any one of claims 140 to 150, wherein the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

152. The heterodimer complex according to any one of claims 140 to 151, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

153. The IL-12p35 subunit is i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Sequence ID 118 (IL-12p35 (E38Q)), vii) Sequence ID 119 (IL-12p35 (N151D)), viiii) Sequence ID 120 (IL-12p35 ( Q135E), ix) SEQ ID NO: 121 (IL-12p35 (Q35D)), x) SEQ ID NO: 122 (IL-12p35 (Q146E)), xi) SEQ ID NO: 123 (IL-12p35 (N76D)), xi) SEQ ID NO: 124 (IL-12p35 (E162Q)), xiiii) SEQ ID NO: 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) SEQ ID NO: XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL- 12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxv ii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)),xxxiv) Sequence ID XXX (IL-12p35 (M97A)), xxxv) Sequence ID XXX (IL-12p35 (L89A)), xxxvi) Sequence ID XXX (IL-12p35 (L124A)), xxxvii) Sequence ID XXX (IL-12p35 (M125A)), xxxviiii) Sequence ID XXX (IL-12p35 (L75A)), xxxiv) Sequence ID XXX (IL-12p35 (I171A)), xxxv) Sequence number Number XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Distribution, A heterodimer complex according to any one of claims 140 to 152, having a polypeptide sequence selected from the group consisting of sequence number XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)), xliiii) Sequence ID XXX (IL-12p35 (N71D / N195D), xliv) Sequence ID XXX (IL-12p35 (N85D / N195D)), xlv) Sequence ID XXX (IL-12p35 (D55Q)), and xlvi) Sequence ID XXX (IL-12p35 (D55K)).

154. A composition comprising a heterodimer complex according to any one of claims 140 to 153 for use in the treatment of cancer in a subject.

155. One or more nucleic acids encoding a heterodimer complex according to any one of claims 140 to 153.

156. A host cell comprising one or more nucleic acids as described in claim 155.

157. A method for producing a heterodimer complex, wherein the method comprises culturing the host cells described in claim 156 under conditions that produce the heterodimer complex.

158. IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or variant IL-12p40 subunit having modified affinity for the IL-12 receptor complex.

159. The aforementioned variant IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y66, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F1 06, E110, N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q17 2, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R20 8, D209, D214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248 The variant IL-12p40 subunit according to claim 1, having one or more amino acid modifications with amino acid residues selected from the group consisting of Q256, C252, K258, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

160. The variant IL-12p40 subunit is D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, The variant Il-12p40 subunit according to claim 158 or 159, having one or more amino acid substitutions selected from the group consisting of K163E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

161. The aforementioned variant IL-12p40 subunit is N103D / N113D / N200 D / N281D、Q42E / E45Q、E45Q / Q56E、Q4 2E / E59Q、Q56E / E59Q、Q42E / E45Q / Q56 E、E45Q / Q56E / E59Q、E32Q / E59Q、D34N / E59K、D34K / E59K / K99E、E32Q / D34N / E59K / K99E、E32K / D34N / E59K / K99E、D34N / E59Q、E59Q / E187Q、S43E / E59Q、S43K / E49Q、E59Q / K163E、E59Q / K99E、E59Q / K258E、E59Q / K260E、E59K / K99E、D18K / E59K / K99E、E59K / K99E / K264E、E59K / K99Y、E59 Y / K99Y、E59Y / K99E、E45K / E59K / K99E、E59K / K99E / Q144E、E59K / K99E / Q144 K、E59K / K99E / R159E、E59K / K99E / K264E、D18K / E59K / K99E / K264E、DI8K / E 59K / K99E / C252S、D18K / E59K / K99E / C252S / K264E、E59K / K99Y / C252S、E59K / K99E / C252S / K264E、E59K / K99E / C2 52S、N103D / N113D、N103D / N200D、N10 3D / N281D、N113D / N200D、N113D / N281 D、N200D / N281D、N103D / N113D / N200D 、N103D / N113D / N281D、N103D / N200D / N281D、N113D / N200D / N281D、N103Q / N113Q、N103Q / N200Q、N103Q / N281Q、N 113Q / N200Q、N113Q / N281Q、N200Q / N2 81Q、N103Q / N113Q / N200Q、N103Q / N1 13Q / N281Q、N103Q / N200Q / N281Q、N11 3Q / N200Q / N281Q、N103Q / N113Q / N200 Q / N281Q、E59K / K99E / N103Q / C252S / K 264E、E59K / K99E / N113Q / C252S / K264E、E59K / K99E / N200Q / C252S / K264E、 E59K / K99E / N281Q / C252S / K264E、E59 K / K99E / N103Q / N113Q / C252S / K264E、E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / C252S / K264 E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / N200Q / C25 A variant IL-12p40 subunit according to any one of claims 158 to 160, having an amino acid substitution selected from the group consisting of 2S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

162. The variant IL-12p40 subunits are i) SEQ ID NO: 57 (IL-12p40(N103D)), ii) SEQ ID NO: 58 (IL-12p40(N113D)), iii) SEQ ID NO: 59 (IL-12p40(N200D)), iv) SEQ ID NO: 60 (IL-12p40(N281D)), v) SEQ ID NO: 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) SEQ ID NO: 62 (IL-12p40(Q42E)), vii) SEQ ID NO: 63 (IL-12p40(E45Q)), viiii) SEQ ID NO: 64 (IL-12p40(Q56E)), ix) SEQ ID NO: 65 (IL-12p40(E59Q) )), x) SEQ ID NO: 66 (IL-12p40 (D62N)), xi) SEQ ID NO: 67 (IL-12p40 (Q42E / E45Q)), xi) SEQ ID NO: 68 (IL-12p40 (E45Q / Q56E)), xiiii) SEQ ID NO: 69 (IL-12p40 (Q42E / E59Q)), xiv) SEQ ID NO: 70 (IL-12p40 (Q56E / E59Q)), xv) SEQ ID NO: 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) SEQ ID NO: 72 (IL-12p40 (E45Q / Q56E / E59Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 ( E73Q), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q)), xxvi) SEQ ID NO: 82 (IL-12p40 (Q256N)), xxvii) SEQ ID NO: 83 (IL- 12p40 (E299Q), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)), xxxiv) Sequence ID 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxv i) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) Sequence ID 206 (IL-12p40 (E32Q / E59Q)), xli) Sequence ID 207 (IL-12p40 (D34N / E59Q)), xlii) Sequence ID 208 (IL-12p40 (E59Q / E187Q)), xliiii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvii) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), l) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)),lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E 59K / K99E / R159E)), lx) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 ( E59K / K99Y / C252S), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D) ), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)),lxxvii) Sequence ID XXX (, IL-12p40(N103Q)), lxxviiii) Sequence ID XXX (IL-12p40(N113Q)), lxxix) Sequence ID XXX (IL-12p40(N200Q)), lxxx) Sequence ID XXX (IL-12p40(N281Q)), lxxxi) Sequence ID XXX (IL-12p40(N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40(N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40(N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL -12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX ( IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)),ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N10 3Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX (I L-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N2 A variant IL-12p40 according to any one of claims 158 to 161, having a polypeptide sequence selected from the group consisting of (00Q / N281Q / C252S / K264E)) and cix) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

163. A composition comprising the variant IL-12p40 subunit according to any one of claims 158 to 162.

164. A nucleic acid encoding the variant IL-12p40 subunit according to any one of claims 158 to 162.

165. A host cell containing the nucleic acid according to claim 164.

166. A method for manufacturing a variant IL-12p40 subunit, wherein the method is A method comprising culturing the host cells according to claim 165 under conditions that produce the variant IL-12p40 subunit.

167. IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or variant IL-12p35 subunit having altered affinity for the IL-12 receptor complex.

168. The variant IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q1 The variant IL-12p35 subunit according to claim 167, having one or more amino acid modifications as amino acid residues selected from the group consisting of 46, N151, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

169. The variant IL-12p35 subunit according to claim 167 or 168, wherein the variant IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

170. The aforementioned variant IL-12p35 subunits are N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N A variant IL-12p35 subunit according to any one of claims 167 to 169, having an amino acid substitution selected from the group consisting of 71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

171. The variant IL-12p35 subunit according to any one of claims 167 to 170, wherein the variant IL-12p35 subunit has amino acid substitutions N71D / N85D / N195D.

172. The aforementioned variant IL-12p35 subunit is i) SEQ ID NO: 113 (IL-12p35 (N71D)), ii) SEQ ID NO: 114 (IL-12p35 (N85D)), iii) SEQ ID NO: 115 (IL-12p35 (N195D)), iv) SEQ ID NO: 116 (IL-12p35 (N71D / N85D / N195D)), v) SEQ ID NO: 117 (IL-12p35 (E153Q)), vi) SEQ ID NO: 118 (IL-12p35 (E38Q)), vii) SEQ ID NO: 119 (IL-12p35 (N151D)), viiii) SEQ ID NO: 120 (IL- 12p35 (Q135E), ix) Sequence ID 121 (IL-12p35 (Q35D)), x) Sequence ID 122 (IL-12p35 (Q146E)), xi) Sequence ID 123 (IL-12p35 (N76D)), xi) Sequence ID 124 (IL-12p35 (E162Q)), xiiii) Sequence ID 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) Sequence ID XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), x vii) IL-12p35 (Q130E), xviiii) IL-12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) SEQ ID NO: XXX (IL-12p35 (F96A)), xxxiv) SEQ ID NO: XXX (IL-12p35 (M97A)), xxxv) SEQ ID NO: XXX (IL-12p35 (L89A)), xxxvi) SEQ ID NO: XXX (IL-12p35 (L124A)), xxxvii) SEQ ID NO: XXX (IL-12p35 (M125A)), xxxviiii) SEQ ID NO: XXX (IL-12p35 (L75A)), xxxiv) SEQ ID NO: XXX (IL-12p35 (I171A)), xxxv) SEQ ID NO: XXX (IL-12p35 (N71Q) )), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Sequence ID XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)),A variant IL-12p35 subunit according to any one of claims 167 to 171, having a polypeptide sequence selected from the group consisting of xliiii) SEQ ID NO: XXX (IL-12p35 (N71D / N195D), xliv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

173. A composition comprising the variant IL-12p35 subunit according to any one of claims 167 to 172.

174. A nucleic acid encoding the variant IL-12p35 subunit according to any one of claims 167 to 172.

175. A host cell containing the nucleic acid according to claim 174.

176. A method for producing a variant IL-12p35 subunit, the method comprising culturing the host cells described in claim 175 under conditions that produce the variant IL-12p35 subunit.

177. It is a heterodimer protein, a) A first fusion protein containing an IL-12p40 subunit domain covalently bonded to the first Fc domain, b) A second fusion protein comprising an IL-12p35 subunit domain covalently bonded to a second Fc domain, The first and second Fc domains are, according to EU numbering, L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K A heterodimeric protein, which is a set of amino acid substitutions selected from the group consisting of 370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q; T366S / L368A / Y407V and T366W; and T366S / L368A / Y407V / Y349C and T366W / S354C.

178. The heterodimer protein according to claim 177, wherein the IL-12p40 subunit domain is bound to the N-terminus of the first Fc domain, and the IL-12p35 subunit domain is bound to the N-terminus of the second Fc domain.

179. The heterodimer protein according to claim 177, wherein the IL-12p40 subunit domain is bound to the C-terminus of the first Fc domain, and the IL-12p35 subunit domain is bound to the C-terminus of the second Fc domain.

180. The heterodimer protein according to any one of claims 177 to 179, wherein the Il-12p40 subunit domain is bound to the first Fc domain using a domain linker, and the IL-12p35 subunit domain is bound to the second Fc domain using a domain linker.

181. The heterodimer protein according to any one of claims 177 to 180, wherein the first and second Fc domains comprise IgG1 hinge-CH2-CH3.

182. The heterodimer protein according to claim 177, wherein the first fusion protein comprises an IL-12p40 subunit domain-domain linker-hinge-CH2-CH3 from the N-terminus to the C-terminus, and the second fusion protein comprises an IL-12p35 subunit domain-domain linker-hinge-CH2-CH3 from the N-terminus to the C-terminus.

183. The heterodimer protein according to claim 177, wherein the first fusion protein comprises a hinge-CH2-CH3-domain linker-IL-12p40 subunit domain from the N-terminus to the C-terminus, and the second fusion protein comprises a hinge-CH2-CH3-domain linker-IL-12p35 subunit domain from the N-terminus to the C-terminus.

184. It is a heterodimer protein, a) The first FC domain and b) A fusion protein, i) The second FC domain, ii) IL-12p40 subunit domain and iii) A fusion protein comprising the IL-12p35 subunit domain, A heterodimer protein in which the first and second Fc domains are sets of amino acid substitutions selected from the group consisting of L368D / K370S and S364K; L368D / K370S and S364K / E357L; L368D / K370S and S364K / E357Q; T411E / K360E / Q362E and D401K; L368E / K370S and S364K; K370S and S364K / E357Q; T366S / L368A / Y407V and T366W; and T366S / L368A / Y407V / Y349C and T366W / S354C, according to EU numbering.

185. The heterodimer protein according to claim 184, wherein the fusion protein comprises, from the N-terminus to the C-terminus, an IL-12p40 subunit domain - domain linker - IL-12p35 subunit domain - domain linker - hinge - CH2 - CH3.

186. The heterodimer protein according to claim 184, wherein the fusion protein comprises, from the N-terminus to the C-terminus, IL-12p35 subunit domain - domain linker - IL-12p40 subunit domain - domain linker - hinge - CH2 - CH3.

187. The heterodimer protein according to claim 184, wherein the fusion protein comprises a hinge-CH2-CH3 domain linker-IL-12p35 subunit domain-domain linker-IL-12p40 subunit domain from the N-terminus to the C-terminus.

188. The heterodimer protein according to claim 184, wherein the fusion protein comprises a hinge-CH2-CH3 domain linker IL-12p40 subunit domain-domain linker-IL-12p35 subunit domain from the N-terminus to the C-terminus.

189. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E) ), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), iii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59 Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) SEQ ID NO: 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) SEQ ID NO: 206 (IL-12p40 (E32Q / E59Q)), xli) SEQ ID NO: 207 (IL-12p40 (D34N / E59Q)), xlii) SEQ ID NO: 208 (IL-12p40 (E59Q / E187Q)) , xliiii) Sequence ID 209 (IL-12p40 (S43E / E59Q)), xliv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviiii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E5, 9K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), li) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K 99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144K)), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C25 2S)), lxiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N 103D / N200D), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)),lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)), lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)), lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N 113Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), lxxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34 K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)),xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)), c) Sequence ID, XXX (IL-12p40 (E59K / K99E / N281Q / C252S / K264E)), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), c iii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C 252S / K264E)), cvi) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID XXX (IL-12p40 ( A heterodimer protein according to any one of claims 177 to 188, having a sequence selected from the group consisting of E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E)), and cix) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

190. The IL-12p35 subunit is i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Sequence ID 118 (IL-12p35 (E38Q)), vii) Sequence ID 119 (IL-12p35 (N151D)), viiii) Sequence ID 120 (IL-12p35 ( Q135E), ix) SEQ ID NO: 121 (IL-12p35 (Q35D)), x) SEQ ID NO: 122 (IL-12p35 (Q146E)), xi) SEQ ID NO: 123 (IL-12p35 (N76D)), xi) SEQ ID NO: 124 (IL-12p35 (E162Q)), xiiii) SEQ ID NO: 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) SEQ ID NO: XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL- 12p35 (N136D), xix) IL-12p35 (E143Q), xx) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxv ii) Sequence ID 230 (IL-12p35 (N151D / K158E)), xxviiii) Sequence ID 231 (IL-12p35 (E79Q / N151D)), xxix) Sequence ID 232 (IL-12p35 (D55Q / N151D)), xxx) Sequence ID 233 (IL-12p35 (N136D / N151D)), xxxi) Sequence ID 234 (IL-12p35 (N21D / N151D)), xxxii) Sequence ID 235 (IL-12p35 (E143Q / N151D)), xxxiii) Sequence ID XXX (IL-12p35 (F96A)),xxxiv) SEQ ID NO: XXX (IL-12p35 (M97A)), xxxv) SEQ ID NO: XXX (IL-12p35 (L89A)), xxxvi) SEQ ID NO: XXX (IL-12p35 (L124A)), xxxvii) SEQ ID NO: XXX (IL-12p35 (M125A)), xxxviiii) SEQ ID NO: XXX (IL-12p35 (L75A)), xxxiv) SEQ ID NO: XXX (IL-12 p35 (I171A)), xxxv) Sequence ID XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)), xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q, A heterodimer protein according to any one of claims 177 to 188, having a sequence selected from the group consisting of )), lx) SEQ ID NO: XXX (IL-12p35 (N85Q / N195Q), lxi) SEQ ID NO: XXX (IL-12p35 (N71Q / N85Q / N195Q)), lxi) SEQ ID NO: XXX (IL-12p35 (N71D / N85D)), lxiiii) SEQ ID NO: XXX (IL-12p35 (N71D / N195D), lxiv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), lxv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and lxvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

191. The heterodimer Fc fusion protein according to any one of claims 177 to 190, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.

192. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an IL-12p40 subunit domain - an arbitrary first domain linker - an IL-12p35 subunit domain - an arbitrary second domain linker - an Fc domain.

193. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an IL-12p35 subunit domain - an arbitrary first domain linker - an IL-12p40 subunit domain - an arbitrary second domain linker - an Fc domain.

194. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an Fc domain - an arbitrary first domain linker - an IL-12p40 subunit domain - an arbitrary second domain linker - an IL-12p35 subunit domain.

195. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an Fc domain – an arbitrary first domain linker – an IL-12p35 subunit domain – an arbitrary second domain linker – an IL-12p40 subunit domain.

196. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an IL-12p40 subunit domain - first domain linker - IL-12p35 subunit domain - second domain linker - Fc domain.

197. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an IL-12p35 subunit domain - first domain linker - IL-12p40 subunit domain - second domain linker - Fc domain.

198. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each comprising, from the N-terminus to the C-terminus, an Fc domain - first domain linker - IL-12p40 subunit domain - second domain linker - IL-12p35 subunit domain.

199. A homodimer Fc fusion protein comprising a first monomer and a second monomer, each having, from N-terminus to C-terminus, an Fc domain - first domain linker - IL-12p35 subunit domain - second domain linker - IL-12p40 subunit domain A homodimeric Fc fusion protein containing this protein.

200. The homodimerized Fc fusion protein according to claims 192 to 199, wherein the modification that promotes homodimerization of the Fc domain is a set of amino acid substitutions selected from the group consisting of L368D / K370S;S364K;S364K / E357L;S364K / E357Q;T411E / K360E / Q362E;D401K;T366S / L368A / Y407V;T366W;T366S / L368A / Y407V / Y349C; and T366W / S354C, according to EU numbering.

201. The homodimeric Fc fusion protein according to claims 192-200, wherein the first domain linker and the second domain linker have the same amino acid sequence.

202. The homodimeric Fc fusion protein according to any one of claims 192 to 201, wherein the Fc domain has an additional set of amino acid substitutions including Q295E / N384D / Q418E / N421D according to EU numbering.

203. The homodimeric Fc fusion protein according to any one of claims 192 to 202, wherein the Fc domain has an additional set of amino acid substitutions selected from the group consisting of G236R / L328R, E233P / L234V / L235A / G236_ / S239K, E233P / L234V / L235A / G236_ / S239K / A327G, E233P / L234V / L235A / G236_, and E233P / L234V / L235A / G236_ / S267K, according to EU numbering.

204. The homodimeric Fc fusion protein according to any one of claims 192 to 203, wherein the IL-12p40 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 3 (human IL-12 subunit beta (IL-12p40) precursor sequence) and SEQ ID NO: 4 (human IL-12 subunit beta (IL-12p40) mature form sequence), and the IL-12p35 subunit has a polypeptide sequence selected from the group consisting of SEQ ID NO: 1 (human IL-12 subunit alpha (IL-12p35) precursor sequence) and SEQ ID NO: 2 (human IL-12 subunit alpha (IL-12p35) mature form sequence).

205. The homodimeric Fc fusion protein according to any one of claims 192 to 204, wherein each of the Fc domains further comprises the amino acid substitution M428L / N424S.

206. The homodimer Fc fusion protein according to any one of claims 192 to 205, wherein the IL-12p40 subunit is a variant IL-12p40 subunit and / or the IL-12p35 subunit is a variant IL-12p35 subunit.

207. The homodimeric Fc fusion protein according to any one of claims 192 to 206, wherein the IL-12p40 subunit is a variant IL-12p40 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex, and / or the IL-12p35 subunit is a variant IL-12p35 subunit having a modified affinity for IL-12 receptor subunit beta-1 (IL-12Rβ1), IL-12 receptor subunit beta-2 (IL-12Rβ2), and / or IL-12 receptor complex.

208. The IL-12p40 subunit is E3, D7, E12, D14, W15, P17, D18, A19, P20, G21, E22, M23, D29, E32, E33, D34, L40, D41, Q42, S43, E45, L47, T54, I55, Q56, K58, E59, F60, G61, D62, Q65, Y6 6, E73, K84, E86, D87, G88, I89, W90, D93, D97, K99, E100, K102, N103, K104, F106, E110 , N113, Y114, D129, D142, Q144, E156, R159, D161, N162, K163, D166, D170, Q172, D174, A176, C177, P178, A179, A180, E181, S183, P185, E187, N200, S204, F206, R208, D209, D 214, N218, Q220, N226, Q229, E231, E235, T242, P243, S245, Y246, F247, S248, C252S, Q A homodimer Fc fusion protein according to any one of claims 192 to 207, having one or more amino acid modifications with amino acid residues selected from the group consisting of 256, K158, K260, E262, K264, D265, D270, N281, Q289, D290, R291, Y292, Y293, and E299.

209. The IL-12p40 subunits are D18N, D18K, E32Q, E33Q, D34N, D34K, Q42E, S43E, S43K, E45Q, Q56E, E59Q, E59K, D62N, E73Q, D87N, K99E, K99Y, E100Q, N103D, N103Q, N113D, N113Q, Q144E, D161N, R159E, K16 A homodimer Fc fusion protein according to any one of claims 192 to 208, having one or more amino acid substitutions selected from the group consisting of 3E, E187Q, N200D, N200Q, N218Q, Q229E, E235Q, C252S, Q256N, K258E, K260E, E262Q, K264E, N281D, N281Q, and E299Q.

210. The IL-12p40 subunits are N103D / N113D / N200D / N281D, Q42E / E45Q, E45Q / Q56E, Q42E / E59Q, Q56E / E59Q, Q42E / E45Q / Q56E, E45Q / Q56E / E59Q, E32Q / E59Q, D34N / E59K, D34N / E59K / K99E, D34K / E59K / K99E, E32Q / D34N / E59K / K99E, E32K / D34N / E59K / K99E, D34N / E59Q, E59Q / E187Q, S43E / E59Q, S43K / E49Q, E59Q / K 163E, E59Q / K99E, E59Q / K258E, E59Q / K260E, E59K / K99E, D18K / E59K / K99E, E59K / K99E / K264E, E59K / K99Y, E59Y / K99Y, E59Y / K99E, E45K / E59K / K99E, E5 9K / K99E / Q144E, E59K / K99E / Q144K, E59K / K99E / R159E, E59K / K99E / K264E, D18K / E59K / K99E / K264E, DI8K / E59K / K99E / C252S, D18K / E59K / K99E / C252S / K264E, E59K / K99Y / C252S, E59K / K99E / C252S / K264E, E59K / K99E / C252S, N1 03D / N113D, N103D / N200D, N103D / N281D, N113D / N200D, N113D / N281D, N200D / N281D, N103D / N113D / N200D, N103D / N113D / N281D, N103D / N200D / N281D, N 113D / N200D / N281D, N103Q / N113Q, N103Q / N200Q, N103Q / N281Q, N113Q / N200 Q, N113Q / N281Q, N200Q / N281Q, N103Q / N113Q / N200Q, N103Q / N113Q / N281Q, N103Q / N200Q / N281Q, N113Q / N200Q / N281Q, N103Q / N113Q / N200Q / N281Q, E59 K / K99E / N103Q / C252S / K264E, E59K / K99E / N113Q / C252S / K264E, E59K / K99E / N200Q / C252S / K264E, E59K / K99E / N281Q / C252S / K264E, E59K / K99E / N103Q / N113Q / C252S / K264E, E59K / K99E / N103Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N 200Q / C252S / K264E, E59K / K99E / N113Q / N281Q / C252S / K264E, E59K / K99E / N200Q / N281Q / C252S / K264E, E59K / K99E / N103Q / N11 A homodimer Fc fusion protein according to any one of claims 192 to 209, having amino acid substitutions selected from the group consisting of 3Q / N200Q / C252S / K264E, E59K / K99E / N103Q / N200Q / N281Q / C252S / K264E, E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E, and E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E.

211. The IL-12p40 subunits are: i) Sequence ID 57 (IL-12p40(N103D)), ii) Sequence ID 58 (IL-12p40(N113D)), iii) Sequence ID 59 (IL-12p40(N200D)), iv) Sequence ID 60 (IL-12p40(N281D)), v) Sequence ID 61 (IL-12p40(N103D / N113D / N200D / N281D)), vi) Sequence ID 62 (IL-12p40(Q42E)), vii) Sequence ID 63 (IL-12p40(E45Q)), viiii) Sequence ID 64 (IL-12p40(Q56E) ), ix) Sequence ID 65 (IL-12p40 (E59Q)), x) Sequence ID 66 (IL-12p40 (D62N)), xi) Sequence ID 67 (IL-12p40 (Q42E / E45Q)), xii) Sequence ID 68 (IL-12p40 (E45Q / Q56E)), iii) Sequence ID 69 (IL-12p40 (Q42E / E59Q)), xiv) Sequence ID 70 (IL-12p40 (Q56E / E59Q)), xv) Sequence ID 71 (IL-12p40 (Q42E / E45Q / Q56E)), xvi) Sequence ID 72 (IL-12p40 (E45Q / Q56E / E59 Q)), xvii) SEQ ID NO: 73 (IL-12p40 (D161N)), xviiii) SEQ ID NO: 74 (IL-12p40 (E73Q)), xix) SEQ ID NO: 75 (IL-12p40 (Q144E)), xx) SEQ ID NO: 76 (IL-12p40 (E262Q)), xxi) SEQ ID NO: 77 (IL-12p40 (E100Q)), xxii) SEQ ID NO: 78 (IL-12p40 (D18N)), xxiii) SEQ ID NO: 79 (IL-12p40 (E33Q)), xxiv) SEQ ID NO: 80 (IL-12p40 (Q229E)), xxv) SEQ ID NO: 81 (IL-12p40 (E235Q), xxvi) Sequence ID 82 (IL-12p40 (Q256N)), xxvii) Sequence ID 83 (IL-12p40 (E299Q)), xxviiii) Sequence ID 84 (IL-12p40 (D87N)), xxix) IL-12p40 (E32Q), xxx) IL-12p40 (D34N), xxxi) IL-12p40 (S43E), xxxii) IL-12p40 (S43K), xxxiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / N200Q / N281Q / C252S / K264E)),xxxiv) SEQ ID NO: 205 (IL-12p40 (E59K)), xxxv) IL-12p40 (K99E), xxxvi) IL-12p40 (K163E), xxxvii) IL-12p40 (E187Q), xxxviiii) IL-12p40 (K258E), xxxix) IL-12p40 (K260E), xl) SEQ ID NO: 206 (IL-12p40 (E32Q / E59Q)), xli) SEQ ID NO: 207 (IL-12p40 (D34N / E59Q)), xliii) SEQ ID NO: 208 (IL-12p40 (E59Q / E187Q)), xliiii) SEQ ID NO: 209 (IL-12 p40 (S43E / E59Q)), xlv) Sequence ID 210 (IL-12p40 (S43K / E49Q)), xlv) Sequence ID 211 (IL-12p40 (E59Q / K163E)), xlvi) Sequence ID 212 (IL-12p40 (E59Q / K99E)), xlvii) Sequence ID 213 (IL-12p40 (E59Q / K258E)), xlviii) Sequence ID 214 (IL-12p40 (E59Q / K260E)), xlix) Sequence ID XXX (IL-12p40 (D34N / E59K)), l) Sequence ID XXX (IL-12p40 (E59K / K99E)), l) Distribution, Column number XXX (IL-12p40 (D18K / E59K / K99E)), lii) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), liiii) Sequence ID XXX (IL-12p40 (E59K / K99Y)), liv) Sequence ID XXX (IL-12p40 (E59Y / K99E)), lv) Sequence ID XXX (IL-12p40 (E45K / E59K / K99E)), lvi) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E / Q144E)), lvii) Sequence ID XXX (IL-12p40 (E59K / K99E) 9E / Q144K), lviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / R159E)), lix) Sequence ID XXX (IL-12p40 (E59K / K99E / K264E)), lx) Sequence ID XXX (IL-12p40 (D18K / E59K / K99E / K264E)), lxi) Sequence ID XXX (IL-12p40 (C252S)), lxi) Sequence ID XXX (IL-12p40 (DI8K / E59K / K99E / C252S)), lxiiii) Sequence ID XXX (IL-12p40 (D18K / E59K / K99 E / C252S / K264E)), lxiv) Sequence ID XXX (IL-12p40 (E59K / K99Y / C252S)), lxv) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S / K264E)), lxvi) Sequence ID XXX (IL-12p40 (E59K / K99E / C252S)), lxvii) Sequence ID XXX (IL-12p40 (N103D / N113D)), lxviiii) Sequence ID XXX (IL-12p40 (N103D / N200D)), lxix) Sequence ID XXX (IL-12p40 (N103D / N281D)), lxx) Sequence ID XXX (IL-12p40 (N113D / N200D)), lxxi) Sequence ID XXX (IL-12p40 (N113D / N281D)), lxxii) Sequence ID XXX (IL-12p40 (N200D / N281D)), lxxiiii) Sequence ID XXX (IL-12p50 (N103D / N113D / N200D)), lxxiv) Sequence ID XXX (IL-12p40 (N103D / N113D / N281D)), lxxv) Sequence ID XXX (IL-12p40 (N103D / N200D / N281D)),lxxvi) Sequence ID XXX (IL-12p40 (N113D / N200D / N281D)), lxxvii) Sequence ID XXX (IL-12p40 (N103Q)), lxxviiii) Sequence ID XXX (IL-12p40 (N113Q)), lxxix) Sequence ID XXX (IL-12p40 (N200Q)), lxxx) Sequence ID XXX (IL-12p40 (N281Q)), lxxxi) Sequence ID XXX (IL-12p40 (N103Q / N113Q)), lxxxii) Sequence ID XXX (IL-12p40 (N103Q / N200Q)), lxxxiii) Sequence ID XXX (IL-12p40 (N103Q / N281Q)), lxxxiv) Sequence ID XXX (IL-12p40 (N113Q / N200Q)), lxxxv) Sequence ID XXX (IL-12p40 (N113Q / N281Q)), lxxxvi) Sequence ID XXX (IL-12p40 (N200Q / N281Q)), lxxxvii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q)), lxxxviiii) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N281Q)), l xxxix) Sequence ID XXX (IL-12p40 (N103Q / N200Q / N281Q)), xc) Sequence ID XXX (IL-12p40 (N113Q / N200Q / N281Q)), xci) Sequence ID XXX (IL-12p40 (N103Q / N113Q / N200Q / N281Q)), xcii) Sequence ID XXX (IL-12p40 (D34N / E59K / K99E)), xciiii) Sequence ID XXX (IL-12p40 (D34K / E59K / K99E)), xciv) Sequence ID XXX (IL-12p40 (E32Q / D34N / E59 K / K99E)), xcv) Sequence ID XXX (IL-12p40 (E32K / D34N / E59K / K99E)), xcvi) Sequence ID XXX (IL-12p40 (E59Y / K99Y)), xcvii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / C252S / K264E)), xcviiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / C252S / K264E)), xcix) Sequence ID XXX (IL-12p40 (E59K / K99E / N200Q / C252S / K264E)),c) Sequence ID XXX (IL-12p40(E59K / K99E / N281Q / C252S / K264E, )), ci) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N113Q / C252S / K264E)), cii) Sequence ID XXX (IL-12p40 (E59K / K99E / N103Q / N200Q / C252S / K264E)), ciiii) Sequence ID XXX (IL-12p40 (E59K / K99E / N 103Q / N281Q / C252S / K264E)), civ) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / C252S / K264E)), cv) Sequence ID XXX (IL-12p40 (E59K / K99E / N113Q / N281Q / C252S / K264E)), cvi) Sequence ID XXX ( IL-12p40(E59K / K99E / N200Q / N281Q / C252S / K264E)), cvii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N113Q / N200Q / C252S / K264E)), cviiii) Sequence ID XXX(IL-12p40(E59K / K99E / N103Q / N2 A homodimer Fc fusion protein according to any one of claims 192 to 210, having a polypeptide sequence selected from the group consisting of (00Q / N281Q / C252S / K264E)) and cix) SEQ ID NO XXX (IL-12p40 (E59K / K99E / N113Q / N200Q / N281Q / C252S / K264E)).

212. The IL-12p35 subunit is Q20, N21, Q35, E38, S44, E45, E46, H49, K54, D55, T59, V60, E61, C63, L64, P65, E67, L68, N71, S73, C74, L75, N76, E79, N85, L89, F96, M97, L124, M125, Q130, Q135, N136, E143, Q146, N1 A homodimer Fc fusion protein according to any one of claims 192 to 211, having one or more amino acid modifications as amino acid residues selected from the group consisting of 51, E153, K158, E162, E163, D165, I171, R181, I182, R183, V185, T186, D188, R189, V190, S192, Y193, N195, and A196.

213. The homodimer Fc fusion protein according to any one of claims 192 to 212, wherein the IL-12p35 subunit has one or more amino acid substitutions selected from the group consisting of N21D, Q35D, E38Q, D55Q, D55K, N71D, N71Q, L75A, N76D, E79Q, N85D, N85Q, L89A, F96A, M97A, L124A, M125A, Q130E, Q135E, N136D, E143Q, Q146E, N151D, N151K, E153K, E153Q, K158E, E162Q, E163Q, D165N, I171A, N195D, and N195Q.

214. The homodimer Fc fusion protein according to any one of claims 192 to 213, wherein the IL-12p35 subunit has an amino acid substitution selected from the group consisting of N71D / N85D / N195D, N151D / E153Q, N151D / D165N, Q130E / N151D, N151D / K158E, E79Q / N151D, D55Q / N151D, N136D / N151D, N21D / N151D, E143Q / N151D, N71Q / N85Q, N71Q / N195Q, N85Q / N195Q, N71Q / N85Q / N195Q, N71D / N85D, N71D / N195D, and N85D / N195D.

215. The IL-12p35 subunits are: i) Sequence ID 113 (IL-12p35 (N71D)), ii) Sequence ID 114 (IL-12p35 (N85D)), iii) Sequence ID 115 (IL-12p35 (N195D)), iv) Sequence ID 116 (IL-12p35 (N71D / N85D / N195D)), v) Sequence ID 117 (IL-12p35 (E153Q)), vi) Sequence ID 118 (IL-12p35 (E38Q)), vii) Sequence ID 119 (IL-12p35 (N151D)), viiii) Sequence ID 120 (IL-12p35 (Q135E)), ix) Sequence ID 121 (IL-12p35 (Q35D)), x ) Sequence ID 122 (IL-12p35 (Q146E)), xi) Sequence ID 123 (IL-12p35 (N76D)), xii) Sequence ID 124 (IL-12p35 (E162Q)), xiiii) Sequence ID 125 (IL-12p35 (E163Q)), xiv) IL-12p35 (N21D), xv) Sequence ID XXX (IL-12p35 (D55Q)), xvi) IL-12p35 (E79Q), xvii) IL-12p35 (Q130E), xviiii) IL-12p35 (N136D), xix) IL-12p35 (E143Q), xx ) Sequence ID 227 (IL-12p35 (N151K)), xxi) Sequence ID 226 (IL-12p35 (E153K)), xxii) IL-12p35 (K158E), xxiii) IL-12p35 (D165N), xxiv) Sequence ID 225 (IL-12p35 (N151D / E153Q)), xxv) Sequence ID 228 (IL-12p35 (N151D / D165N)), xxvi) Sequence ID 229 (IL-12p35 (Q130E / N151D)), xxvii) Sequence ID 230 (IL-12p35 (N151D / K158E)), x xviiii) SEQ ID NO: 231 (IL-12p35 (E79Q / N151D)), xxx) SEQ ID NO: 232 (IL-12p35 (D55Q / N151D)), xxx) SEQ ID NO: 233 (IL-12p35 (N136D / N151D)), xxxi) SEQ ID NO: 234 (IL-12p35 (N21D / N151D)), xxxii) SEQ ID NO: 235 (IL-12p35 (E143Q / N151D)), xxxiii) SEQ ID NO: XXX (IL-12p35 (F96A)), xxxiv) SEQ ID NO: XXX (IL-12p35 (M97A)), xxx v) Sequence ID XXX (IL-12p35 (L89A)), xxxvi) Sequence ID XXX (IL-12p35 (L124A)), xxxvii) Sequence ID XXX (IL-12p35 (M125A)), xxxviiii) Sequence ID XXX (IL-12p35 (L75A)), xxxiv) Sequence ID XXX (IL-12p35 (I171A)), xxxv) Sequence ID XXX (IL-12p35 (N71Q)), xxxvi) Sequence ID XXX (IL-12p35 (N85Q)), xxxvii) Sequence ID XXX (IL-12p35 (N195Q)),xxxviiii) Sequence ID XXX (IL-12p35 (N71Q / N85Q)), xxxix) Sequence ID XXX (IL-12p35 (N71Q / N195Q)), xl) Sequence ID XXX (IL-12p35 (N85Q / N195Q), xli) Sequence ID XXX (IL-12p35 (N71Q / N85Q / N195Q)), xlii) Sequence ID XXX (IL-12p35 (N71D / N85D)), xliiii) A homodimer Fc fusion protein according to any one of claims 192 to 214, having a polypeptide sequence selected from the group consisting of SEQ ID NO: XXX (IL-12p35 (N71D / N195D), xlv) SEQ ID NO: XXX (IL-12p35 (N85D / N195D)), xlv) SEQ ID NO: XXX (IL-12p35 (D55Q)), and xlvi) SEQ ID NO: XXX (IL-12p35 (D55K)).

216. The homodimer Fc fusion protein according to claim 192 or 196, wherein the first and second monomers are XENP31289.

217. The homodimer Fc fusion protein according to any one of claims 192 or 196, wherein the first and second monomers are XENP31291.

218. A composition comprising a homodimeric Fc fusion protein according to any one of claims 192 to 217 for use in the treatment of cancer in a subject.

219. One or more nucleic acids encoding a heterodimer Fc fusion protein according to any one of claims 192 to 217.

220. A host cell comprising one or more nucleic acids as described in claim 219.

221. A method for producing a homodimer Fc fusion protein, wherein the method is the homodimer A method comprising culturing the host cells described in claim 220 under conditions that produce an Fc fusion protein.

222. A method for purifying a homodimer Fc fusion protein according to any one of claims 192 to 217, wherein the method is a) To provide a composition containing the homodimeric Fc fusion protein, b) Packing the composition onto an ion exchange column, c) A method comprising collecting a fraction containing the homodimer Fc fusion protein.

223. A method for treating cancer in a patient who requires treatment for cancer, comprising administering to the patient a therapeutically effective amount of a heterodimer or homodimer Fc fusion protein described in any of the prior claims.

224. The method according to claim 223, further comprising administering a therapeutically effective dose of a checkpoint-blocking antibody.

225. The method according to claim 224, wherein the checkpoint blocking antibody is selected from anti-PD-1 antibody, anti-PD-L1 antibody, anti-TIM3 antibody, anti-TIGIT antibody, anti-LAG3 antibody, and anti-CTLA-4 antibody.

226. The method according to claim 225, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, or pizilizumab.

227. The method according to claim 225, wherein the anti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab.

228. The method according to any one of claims 223 to 227, wherein the patient exhibits an increase in lymphocytes after administration.

229. The method according to any one of claims 223 to 228, wherein the patient shows an increase in peripheral CD8+ T cells after administration.

230. A method for inducing T cell proliferation in a patient who requires T cell proliferation, comprising administering to the patient a therapeutically effective amount of a heterodimer or homodimer Fc fusion protein described in any of the prior claims.

231. The method according to claim 230, further comprising administering a therapeutically effective dose of a checkpoint-blocking antibody.

232. The method according to claim 231, wherein the checkpoint blocking antibody is selected from anti-PD-1 antibody, anti-PD-L1 antibody, anti-TIM3 antibody, anti-TIGIT antibody, anti-LAG3 antibody, and anti-CTLA-4 antibody.

233. The method according to claim 232, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, or pizilizumab.

234. The method according to claim 232, wherein the anti-PD-L1 antibody is atezolizumab, avelumab, or durvalumab.

235. The method according to any one of claims 230 to 234, wherein the T cell proliferation is at least a twofold increase in the number of T cells.

236. The method according to any one of claims 224-229 and 230-235, wherein the IL-12Fc fusion protein is administered prior to the checkpoint inhibitor to increase checkpoint expression before treatment.

237. The method according to any one of claims 226, 228, 229, 233, and 235, wherein the IL-12Fc fusion protein is administered prior to the checkpoint inhibitor to increase checkpoint expression before treatment.

238. The heterodimer Fc fusion protein according to any one of claims 119 to 124, wherein the first and second Fc domains further comprise the amino acid substitution M428L / N424S.