Multispecific molecules binding trailr2 and CDH17 and uses thereof

Abstract

The present application provides multispecific antigen-binding proteins (e.g., antibodies) that specifically bind TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2) and cadherin 17 (CDH17). The application also provides conjugates comprising the multispecific antigen-binding proteins, polynucleotides and recombinant vectors encoding the multispecific antigen-binding proteins, as well as host cells and methods for preparing the multispecific antigen-binding proteins. The application further provides pharmaceutical compositions comprising the multispecific antigen-binding proteins andmethods for treating a disease or disorder using the multispecific antigen-binding proteins.

Description

Attorney Docket No: 260525.000128MULTISPECIFIC MOLECULES BINDING TRAILR2 AND CDH17 AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U. S. Provisional Application No. 63 / 736,465, filed December 19, 2024, and U. S. Provisional Application No. 63 / 787,158, filed April 11, 2025, the disclosures of which are herein incorporated by reference in their entireties.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on December 16, 2025, is named 26052_000128_SL.xml and is 4,491,965 bytes in size.FIELD OF THE INVENTION

[0003] The present invention relates to multispecific antigen-binding proteins, and methods of use thereof. The multispecific antigen-binding proteins, including bispecific molecules, may comprise one or more antigen-binding domains that specifically bind TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2), and one or more antigen-binding domains that specifically bind cadherin 17 (CDH17).BACKGROUND

[0004] The development of therapies promoting the elimination of cancer cells through apoptosis has been investigated for over three decades. Apoptosis controls cellular homeostasis of normal tissue compartments and is tightly regulated on multiple levels. Cancer cells often evade apoptotic cell death through downregulation of pro-apoptotic proteins and / or overexpression of anti-apoptotic proteins which leads to intrinsic resistance to anticancer therapies.

[0005] Induction of apoptosis through TRAILR1 and TRAILR2 agonists or recombinant TRAIL ligand has been explored preclinically and many therapeutic molecules have been tested in clinical trials (Walczak, H. et al. Tumoricidal activity of tumor necrosis factor-related apoptosis-inducing ligand in vivo. Nat Med 5, 157-63 (1999); Ashkenazi, A. et al. Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Invest 104, 155-62 (1999); Ashkenazi, A. & Herbst, R. S. To kill a tumor cell: the potential of proapoptotic receptor agonists. J Clin Invest 118, 1979-90 (2008)). High TRAILR2 expression is observed across multiple indications while TRAILR1 is generally expressed at lower levels (Diaz Arguello, O. A. &1323195900v2Attorney Docket No: 260525.000128Haisma, H. J. Apoptosis-Inducing TNF Superfamily Ligands for Cancer Therapy. Cancers (Basel) 13, (2021)).

[0006] Clustering of multiple TRAILR2 trimeric receptors has been found to correlate with potent pathway activation and current TRAILR2 agonists are attempting to address this need through multivalent targeting of TRAILR2 or optimizing crosslinking of TRAILR2 receptors through targeting arms in an IgG format. Based on recent clinical data (Papadopoulos, K. P. et al. Unexpected hepatotoxicity in a phase I study of TAS266, a novel tetravalent agonistic Nanobody® targeting the DR5 receptor. Cancer Chemother Pharmacol 75, 887-95 (2015)), engagement of TRAILR2 in the liver has been identified as a significant safety issue. There is a need for therapies to avoid accumulation of the drug in the liver.SUMMARY OF THE INVENTION

[0007] As mentioned in the background section above, there is a need in the art to develop molecules which can efficiently target TRAILR2 with minimal or no safety issues. This application provides molecules which can efficiently target TRAILR2 and CDH17 to address this and other related needs. Compositions and methods involving the molecules which efficiently target TRAILR2 and CDH17 are also provided.

[0008] As mentioned in the background section above, there is a need in the art to develop molecules which can efficiently target TRAILR2 with minimal or no safety issues. This application provides molecules which can efficiently target TRAILR2 and CDH17 to address this and other related needs. Compositions and methods involving the molecules which efficiently target TRAILR2 and CDH17 are also provided.

[0009] In one aspect, provided herein is a multispecific antigen-binding protein comprising:a. one or more antigen-binding domains that specifically bind to TNF-related apoptosisinducing ligand receptor 2 (TRAILR2); andb. one or more antigen-binding domains that specifically bind to cadherin 17 (CDH17).

[0010] In some embodiments, the multispecific antigen-binding protein is bivalent, trivalent, tetravalent, pentavalent, hexavalent, septivalent, or octavalent.

[0011] In some embodiments, the multispecific antigen-binding protein comprises at least two, at least three, or at least four anti-TRAILR2 antigen-binding domains, and at least one, at least two, or at least four anti-CDH17 antigen-binding domains.

[0012] In some embodiments, the multispecific antigen-binding protein comprises two anti-TRAILR2 antigen-binding domains, and two or four anti-CDH17 antigen-binding domains.2323195900v2Attorney Docket No: 260525.000128

[0013] In some embodiments, the multispecific antigen-binding protein further comprises one or more antigen-binding domains that binds to another antigen. In some embodiments, the other antigen is serum albumin. In some embodiments, the serum albumin is human serum albumin (HSA).

[0014] In some embodiments, the multispecific antigen-binding protein further comprises an immunoglobulin Fc region.

[0015] In one aspect, provided herein is a multispecific antigen-binding protein comprising a single polypeptide chain, said single polypeptide chain comprisingi) one or more antigen-binding domains that specifically bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2);ii) one or more antigen-binding domain(s) that specifically bind to cadherin-17 (CDH17); and iii) optionally, one or more antigen-binding domains that bind human serum albumin (HSA); and optionally, wherein each of the antigen-binding domains is operably linked to each of the other antigen-binding domains via a linker.

[0016] In some embodiments, the single polypeptide chain comprises three antigen-binding domains (ABDI- ABD3), four antigen-binding domains (ABDI- ABD4), five antigen-binding domains (ABDI- ABD5), or six antigen-binding domains (ABD1-ABD6).

[0017] In some embodiments, the single polypeptide chain comprises three antigen-binding domains, said three antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2; andiii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2.

[0018] In some embodiments, the single polypeptide chain comprises four antigen binding domains, said four antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2; andiv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2.

[0019] In some embodiments, the single polypeptide chain comprises four antigen binding domains, said four antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2; and3323195900v2Attorney Docket No: 260525.000128iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2.

[0020] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0021] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0022] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds HSA; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0023] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds HSA.

[0024] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds TRAILR2;4323195900v2Attorney Docket No: 260525.000128ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds CDH17;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0025] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds TRAILR2;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds CDH17; andv) a fifth antigen-binding domain (ABD5) that specifically binds CDH17.

[0026] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0027] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds HSA.

[0028] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;5323195900v2Attorney Docket No: 260525.000128iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds HSA.

[0029] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds HSA; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0030] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds HSA;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0031] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds HSA;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0032] In one aspect, provided herein is a multispecific antigen-binding protein comprising a first polypeptide chain and a second polypeptide chain, each polypeptide chain comprising:i) one or more antigen-binding domains that specifically bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2),ii) one or more antigen-binding domain that specifically bind to cadherin-17 (CDH17); and6323195900v2Attorney Docket No: 260525.000128iii) an immunoglobulin Fc region; andwherein the first and second polypeptide chains dimerize via the immunoglobulin Fc region to form a dimer, and optionally, wherein each of the antigen-binding domains is operably linked to each of the other antigen-binding domains or the immunoglobulin Fc region via a linker.

[0033] In some embodiments, the first polypeptide chain and / or the second polypeptide chain each comprises two antigen-binding domains (ABD1-ABD2 or ABD3-ABD4), three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), or four antigen-binding domains (ABD1-ABD4 or ABD5-ABD8).

[0034] In some embodiments, the first polypeptide chain and the second polypeptide chain each comprise two antigen-binding domains (ABD1-ABD2 or ABD3-ABD4), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, and (ii) a second antigen-binding domain (ABD2) operably linked to the C-terminus of the first immunoglobulin Fc region; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a third antigen binding domain (ABD3) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, and (ii) a fourth antigen-binding domain (ABD4) operably linked to the C-terminus of the second immunoglobulin Fc region.

[0035] In some embodiments, ABDI and ABD3 specifically binds to CDH17, and ABD2 and ABD4 specifically bind to TRAILR2.

[0036] In some embodiments, ABDI and ABD3 specifically bind to TRAILR2, and ABD2 and ABD4 specifically bind to CDH17.

[0037] In some embodiments, the first polypeptide chain and the second polypeptide chain each comprise three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, (ii) a second antigen-binding domain (ABD2) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker, and (iii) a third antigen-binding domain (ABD3) operably linked to the second antigen-binding domain (ABD2) via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fourth antigen binding domain (ABD4) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, (ii) a fifth antigen-binding domain (ABD5) operably linked to the C-terminus of the second7323195900v2Attorney Docket No: 260525.000128immunoglobulin Fc region via a linker, and (iii) a sixth antigen-binding domain (ABD6) operably linked to the fifth antigen-binding domain (ABD5) via a linker.

[0038] In some embodiments, ABDI and ABD4 specifically bind to CDH17, and ABD2, ABD3, ABD5, and ABD6 specifically bind to TRAILR2.

[0039] In some embodiments, ABDI and ABD4 specifically bind to TRAILR2, and ABD2, ABD3, ABD5, and ABD6 specifically bind to CDH17.

[0040] In some embodiments, the first polypeptide chain and the second polypeptide chain each comprise three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to a second antigen-binding domain (ABD2) via a linker, (ii) said second antigen binding domain (ABD2) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, and (iii) a third antigen-binding domain (ABD3) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fourth antigen binding domain (ABD4) operably linked to a fifth antigen-binding domain (ABD5) via a linker, (ii) said fifth antigen binding domain (ABD5) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, and (iii) a sixth antigen-binding domain (ABD6) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker.

[0041] In some embodiments, ABDI, ABD2, ABD4, and ABD5 specifically bind to CDH17, and ABD3 and ABD6 specifically bind to TRAILR2.

[0042] In some embodiments, ABDI, ABD2, ABD4, and ABD5 specifically bind to TRAILR2, and ABD3 and ABD6 specifically bind to CDH17.

[0043] In some embodiments, the first polypeptide chain and the second polypeptide chain each comprise four antigen-binding domains (ABD1-ABD4 or ABD5-ABD8), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to a second antigen-binding domain (ABD2) via a linker, ii) said second antigen binding domain (ABD2) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, (iii) a third antigen-binding domain (ABD3) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker, and iv) said third antigen-binding domain (ABD3) operably linked to a fourth antigen-binding domain (ABD4) via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fifth antigen binding domain (ABD5) operably linked to a sixth antigen-binding domain (ABD6) via a linker, ii) said8323195900v2Attorney Docket No: 260525.000128sixth antigen binding domain (ABD6) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, (iii) a seventh antigen-binding domain (ABD7) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker, and iv) said seventh antigen-binding domain (ABD7) operably linked to an eighth antigen-binding domain (ABD8) via a linker.

[0044] In some embodiments, ABDI, ABD2, ABD5, and ABD6 specifically bind to CDH17, and ABD3, ABD4, ABD7, and ABD8 specifically bind to TRAILR2.

[0045] In some embodiments, ABDI, ABD2, ABD5, and ABD6 specifically bind to TRAILR2, and ABD3, ABD4, ABD7, and ABD8 specifically bind to CDH17.

[0046] In various embodiments, the one or more anti- TRAILR2 antigen-binding domains comprise a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from: a). (A / V)ASRL(P / V)FNSRSA(I / V)YTDRIYDS (SEQ ID NO: 100);b). AVRRSAWY(S / T)DSIYTVSQYDY (SEQ ID NO: 101);c). NAARSYSR(D / G / N)(G / Y)(E / R)PL(E / K)P(A / D)Y (SEQ ID NO: 102);d). AAASSWSRGG(A / G / I / V)PYGMDY (SEQ ID NO: 103);e). AADS(H / R)FRR(P / Y)(A / T / V)PG(I / Q)QYEY (SEQ ID NO: 104);f). NAA(K / R)SYHRDY(K / S)PL(K / S)(G / P)DY (SEQ ID NO: 105);g). AAAPSFGM(M / R / T)(I / N)PESYVHS (SEQ ID NO: 106);h). AANRGIMSMRLSRYDD (SEQ ID NO: 49);i). T(A / V)GP(A / T)MSYSRGGEF (SEQ ID NO: 107);j). (A / V)ADRGAISRSGAGM(D / N)Y (SEQ ID NO: 108);k). TAGP(A / S)IS(L / Y)SRGGEY (SEQ ID NO: 109);l). A(A / T)NGWGLDP(S / T)TYH(Y / D) (SEQ ID NO: 110);m). SAGWTRRIFQY (SEQ ID NO: 88);n). TAGQSISLSQGGE(H / Y) (SEQ ID NO: 111);o). KAGIRGE(T / V)Y (SEQ ID NO: 112);p). RAYNDGGEY (SEQ ID NO: 2191);q). HS(N / R)WYNL (SEQ ID NO: 2208); andr). (F / M / N)T(A / S)DY,wherein one or more non-alanine residues in the CDR3 sequence are optionally replaced with an alanine, and / or one or more alanine residues in the CDR3 sequence are optionally replaced with a glycine.9323195900v2Attorney Docket No: 260525.000128

[0047] In some embodiments, the CDR3 of the one or more anti-TRAILR2 antigen-binding domains comprises an amino acid sequence selected from:a). (A / G)(A / G)(A / G)(A / S)(A / S)(A / W)(A / S)(A / R)(A / G)(A / G)(A / G / I / V)(A / P)(A / Y)(A / G)(A / M)(A / D) (A / Y);b). (A / T)(A / G / V)(A / G)(A / P)(A / T)(A / M)(A / S)(A / Y)(A / S)(A / R)(A / G)(A / G)(A / E)(A / F);c). (A / R)(A / G)(A / Y)(A / N)(A / D)(A / G)(A / G)(A / E)(A / Y);d). (A / H)(A / S)(A / N / R)(A / W)(A / Y)(A / N)(A / L); ande). (A / F / M / N)(A / T)(A / G / S)(A / D)Y.

[0048] In some embodiments, the CDR3 of the one or more anti-TRAILR2 antigen-binding domains comprises an amino acid sequence selected from: VASRLPFNSRSAIYTDRIYDS (SEQ ID NO: 3);AVRRSAWYSDSIYTVSQYDY (SEQ ID NO: 8); NAARSYSRDYEPLKPDY (SEQ ID NO: 13); NAARSYSRNYEPLKPDY (SEQ ID NO: 16); NAARSYSRGGEPLKPDY (SEQ ID NO: 20); NAARSYSRGGRPLEPAY (SEQ ID NO: 25);AAASSWSRGGVPYGMDY (SEQ ID NO: 30); AADSHFRRYTPGQQYEY (SEQ ID NO: 35); NAAKSYHRDYSPLSPDY (SEQ ID NO: 40); AAAPSFGMRNPESYVHS (SEQ ID NO: 44); AANRGIMSMRLSRYDD (SEQ ID NO: 49);TAGPTMSYSRGGEF (SEQ ID NO: 54); VADRGAISRSGAGMDY (SEQ ID NO: 64); AADRGAISRSGAGMDY (SEQ ID NO: 69); TAGPAISLSRGGEY (SEQ ID NO: 74); TAGPSISYSRGGEY (SEQ ID NO: 78); AANGWGLDPTTYHY (SEQ ID NO: 83); SAGWTRRIFQY (SEQ ID NO: 88); TAGQSISLSQGGEY (SEQ ID NO: 92); KAGIRGEVY (SEQ ID NO: 97); RAYNDGGEY (SEQ ID NO: 2191); HSRWYNL (SEQ ID NO: 2197); FTADY (SEQ ID NO: 2203);GAASSWSRGGVPYGMDY (SEQ ID NO: 2298); AGASSWSRGGVPYGMDY (SEQ ID NO: 2299);AAGSSWSRGGVPYGMDY (SEQ ID NO: 2300); AAAASWSRGGVPYGMDY (SEQ ID NO: 2301);AAASAWSRGGVPYGMDY (SEQ ID NO: 2302); AAASSASRGGVPYGMDY (SEQ ID NO: 2303);AAASSWARGGVPYGMDY (SEQ ID NO: 2304); AAASSWSAGGVPYGMDY (SEQ ID NO: 2305);AAASSWSRAGVPYGMDY (SEQ ID NO: 2306); AAASSWSRGAVPYGMDY (SEQ ID NO: 2307);AAASSWSRGGAPYGMDY (SEQ ID NO: 2308); AAASSWSRGGVAYGMDY (SEQ ID NO: 2309);AAASSWSRGGVPAGMDY (SEQ ID NO: 2310); AAASSWSRGGVPYAMDY (SEQ ID NO: 2311);AAASSWSRGGVPYGADY (SEQ ID NO: 2312); AAASSWSRGGVPYGMAY (SEQ ID NO: 2313);AAASSWSRGGVPYGMDA (SEQ ID NO: 2314); AAGPTMSYSRGGEF (SEQ ID NO: 2315); TGGPTMSYSRGGEF (SEQ ID NO: 2316); TAAPTMSYSRGGEF (SEQ ID NO: 2317); TAGATMSYSRGGEF (SEQ ID NO: 2318);TAGPAMSYSRGGEF (SEQ ID NO: 2319); TAGPTASYSRGGEF (SEQ ID NO: 2320); TAGPTMAYSRGGEF (SEQ ID NO: 2321); TAGPTMSASRGGEF (SEQ ID NO: 2322); TAGPTMSYARGGEF (SEQ ID NO: 2323);TAGPTMSYSAGGEF (SEQ ID NO: 2324); TAGPTMSYSRAGEF (SEQ ID NO: 2325); TAGPTMSYSRGAEF (SEQ ID NO: 2326); TAGPTMSYSRGGAF (SEQ ID NO: 2327); TAGPTMSYSRGGEA (SEQ ID NO: 2328); AAYNDGGEY10323195900v2Attorney Docket No: 260525.000128(SEQ. ID NO: 2329); RGYNDGGEY (SEQ ID NO: 2330); RAANDGGEY (SEQ ID NO: 2331); RAYADGGEY (SEQ ID NO: 2332); RAYNAGGEY (SEQ ID NO: 2333); RAYNDAGEY (SEQ ID NO: 2334); RAYNDGAEY (SEQ ID NO: 2335); RAYNDGGAY (SEQ ID NO: 2336); RAYNDGGEA (SEQ ID NO: 2337); ASRWYNL (SEQ ID NO: 2338); HARWYNL (SEQ ID NO: 2339); HSAWYNL (SEQ ID NO: 2340); HSRAYNL (SEQ ID NO: 2341); HSRWANL (SEQ ID NO: 2342); HSRWYAL (SEQ ID NO: 2343); HSRWYNA (SEQ ID NO: 2344); ATADY (SEQ ID NO: 2345); FAADY (SEQ ID NO: 2346); FTGDY (SEQ ID NO: 2347); and FTAAY (SEQ ID NO: 2348).

[0049] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains further comprise a CDR1 comprising an amino acid sequence selected from:a). GRTFSSNL (SEQ ID NO: 1);b). GGT(F / L)(A / S)N(D / N)G (SEQ ID NO: 113);c). GRTL(D / N / S)(A / D / E)Y(A / G) (SEQ ID NO: 114);d). GRTFS(N / S)YA (SEQ ID NO: 115);e) GRDFSNYV (SEQ ID NO: 33);f). G(L / R)(I / S)FS(D / S)YA (SEQ ID NO: 116);g). GR(A / T)FSTLA (SEQ ID NO: 117);h). GRTFSSDI (SEQ ID NO: 47);i). GRSFGD(D / F / Y)A (SEQ ID NO: 118);j). G(G / R)TLSNYA (SEQ ID NO: 119);k). GRSFGAQGMEG (SEQ ID NO: 72);l). GFTLDLGAYA (SEQ ID NO: 81);m). GFTFGALA (SEQ ID NO: 86);n). GFTLS(G / S)YA (SEQ ID NO: 120);o). GSIFGGYN (SEQ ID NO: 2189);p). G(G / S)NFRILS (SEQ ID NO: 2209); andq). G(F / L)(A / T)F(R / S)(R / S)YA (SEQ ID NO: 2212).

[0050] In some embodiments, the CDR1 of the one or more anti-TRAILR2 antigen-binding domains comprises an amino acid sequence selected from GRTFSSNL (SEQ ID NO: 1); GGTLANNG (SEQ ID NO: 6); GRTLDAYG (SEQ ID NO: 11); GRTLSDYA (SEQ ID NO: 23); GRTFSSYA (SEQ ID NO: 28); GRDFSNYV (SEQ ID NO: 33); GRSFSSYA (SEQ ID NO: 38); GRTFSTLA (SEQ ID NO: 43); GRTFSSDI (SEQ ID NO: 47); GRSFGDFA (SEQ ID NO: 52); GGTLSNYA (SEQ ID NO: 62); GRTLSNYA (SEQ ID NO: 67); GRSFGAQGMEG (SEQ ID NO: 72); GFTLDLGAYA (SEQ ID NO: 81); GFTFGALA (SEQ ID NO: 86); GFTLSGYA (SEQ ID NO: 95); GSIFGGYN (SEQ ID NO: 2189); GSNFRILS (SEQ ID NO: 2195); and GFTFSRYA (SEQ ID NO: 2201).11323195900v2Attorney Docket No: 260525.000128

[0051] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains further comprise a CDR2 comprising an amino acid sequence selected from:a). VSWNGAST (SEQ ID NO: 2);b). DHR(S / T)GT (SEQ ID NO: 121);c). I(N / S)W(N / S / T)G(T / V)(D / G)T (SEQ ID NO: 122);d). LNW(N / S)G(D / E)ST (SEQ ID NO: 123);e). INWAD(E / T)T (SEQ ID NO: 124);f). INWSGG(S / T)T (SEQ ID NO: 125);g). ISWSDMSA (SEQ ID NO: 48);h). I(N / R)W(A / D / T)G(D / N)T (SEQ ID NO: 126);i). ISQ(S / T)S(D / S)T (SEQ ID NO: 127);j). (I / M)KWTGNT (SEQ ID NO: 128);k). ISN(S / T)GTTT (SEQ ID NO: 129);l). ISNDGEHI (SEQ ID NO: 87);m). ISWNGDIT (SEQ ID NO: 91);n). IT(G / S)(A / S)G(G / S)(N / S)T (SEQ ID NO: 130);o). IFISGN(D / N) (SEQ ID NO: 2207);p). (I / L)T(K / M / S)D(D / G)TT (SEQ ID NO: 2210); andq). ISS(A / G / S)(G / S)G(I / Y)(I / T / V) (SEQ ID NO: 2213).

[0052] In some embodiments, the CDR2 of the one or more anti-TRAILR2 antigen-binding domains comprises an amino acid sequence selected from VSWNGAST (SEQ ID NO: 2); DHRSGT (SEQ ID NO: 7); ISWTGVDT (SEQ ID NO: 12); ISWTGTDT (SEQ ID NO: 19); ISWSGVDT (SEQ ID NO: 24); LNWSGEST (SEQ ID NO: 29); INWADET (SEQ ID NO: 34); INWSGGST (SEQ ID NO: 39); ISWSDMSA (SEQ ID NO: 48); IRWTGDT (SEQ ID NO: 53); ISQTSST (SEQ ID NO: 63); ISQSSDT(SEQ ID NO: 68); MKWTGNT (SEQ ID NO: 73);IKWTGNT (SEQ ID NO: 77); ISNTGTTT (SEQ ID NO: 82); ISNDGEHI (SEQ ID NO: 87); ISWNGDIT (SEQ ID NO: 91); ITSAGGST (SEQ ID NO: 96); IFISGNN (SEQ ID NO: 2190); ITSDDTT (SEQ ID NO: 2196); and ISSAGGYI (SEQ ID NO: 2202).

[0053] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise a CDR1 comprising an amino acid sequence selected from SEQ ID NOs: 1, 6, 11, 23, 28, 33, 38, 43, 47, 52, 62, 67, 72, 81, 86, 95, 762-1084, 2189, 2195, 2201, and 2252-2261; a CDR2 comprising an amino acid sequence selected from SEQ ID NOs: 2, 7, 12, 19, 24, 29, 34, 39, 48, 53, 63, 68, 73, 77, 82, 87, 91, 96, 1085-1407, 2190, 2196, 2202, and 2262-2271; and / or a CDR3 comprising an amino acid sequence12323195900v2Attorney Docket No: 260525.000128selected from SEQ ID NOs: 3, 8, 13, 16, 20, 25, 30, 35, 40, 44, 49, 54, 64, 69, 74, 78, 83, 88, 92, 97, 1408-1730, 2191, 2197, 2203, 2272-2277, and 2298-2348.

[0054] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise: i). a CDR1 comprising an amino acid sequence of GRTFSSNL (SEQ ID NO: 1), a CDR2 comprising an amino acid sequence of VSWNGAST (SEQ ID NO: 2), and a CDR3 comprising an amino acid sequence of VASRLPFNSRSAIYTDRIYDS (SEQ ID NO: 3);ii). a CDR1 comprising an amino acid sequence of GGTLANNG (SEQ ID NO: 6), a CDR2 comprising an amino acid sequence of DHRSGT (SEQ ID NO: 7), and a CDR3 comprising an amino acid sequence of AVRRSAWYSDSIYTVSQYDY (SEQ ID NO: 8);iii). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), and a CDR3 comprising an amino acid sequence of NAARSYSRDYEPLKPDY (SEQ ID NO: 13);iv). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), and a CDR3 comprising an amino acid sequence of NAARSYSRNYEPLKPDY (SEQ ID NO: 16);v). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGTDT (SEQ ID NO: 19), and a CDR3 comprising an amino acid sequence of NAARSYSRGGEPLKPDY (SEQ ID NO: 20);vi). a CDR1 comprising an amino acid sequence of GRTLSDYA (SEQ ID NO: 23), a CDR2 comprising an amino acid sequence of ISWSGVDT (SEQ ID NO: 24), and a CDR3 comprising an amino acid sequence of NAARSYSRGGRPLEPAY (SEQ ID NO: 25);vii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);viii). a CDR1 comprising an amino acid sequence of GRDFSNYV (SEQ ID NO: 33), a CDR2 comprising an amino acid sequence of INWADET (SEQ ID NO: 34), and a CDR3 comprising an amino acid sequence of AADSHFRRYTPGQQYEY (SEQ ID NO: 35);ix). a CDR1 comprising an amino acid sequence of GRSFSSYA (SEQ ID NO: 38), a CDR2 comprising an amino acid sequence of INWSGGST (SEQ ID NO: 39), and a CDR3 comprising an amino acid sequence of NAAKSYHRDYSPLSPDY (SEQ ID NO: 40);13323195900v2Attorney Docket No: 260525.000128x). a CDR1 comprising an amino acid sequence of GRTFSTLA (SEQ ID NO: 43), a CDR2 comprising an amino acid sequence of INWSGGST (SEQ ID NO: 39), and a CDR3 comprising an amino acid sequence of AAAPSFGMRNPESYVHS (SEQ ID NO: 44);xi). a CDR1 comprising an amino acid sequence of GRTFSSDI (SEQ ID NO: 47), a CDR2 comprising an amino acid sequence of ISWSDMSA (SEQ ID NO: 48), and a CDR3 comprising an amino acid sequence of AANRGIMSMRLSRYDD (SEQ ID NO: 49);xii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);xiii). a CDR1 comprising an amino acid sequence of GGTLSNYA (SEQ ID NO: 62), a CDR2 comprising an amino acid sequence of ISQTSST (SEQ ID NO: 63), and a CDR3 comprising an amino acid sequence of VADRGAISRSGAGMDY (SEQ ID NO: 64);xiv). a CDR1 comprising an amino acid sequence of GRTLSNYA (SEQ ID NO: 67), a CDR2 comprising an amino acid sequence of ISQSSDT (SEQ ID NO: 68), and a CDR3 comprising an amino acid sequence of AADRGAISRSGAGMDY (SEQ ID NO: 69);xv). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of MKWTGNT (SEQ ID NO: 73), and a CDR3 comprising an amino acid sequence of TAGPAISLSRGGEY (SEQ ID NO: 74);xvi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78);xvii). a CDR1 comprising an amino acid sequence of GFTLDLGAYA (SEQ ID NO: 81), a CDR2 comprising an amino acid sequence of ISNTGTTT (SEQ ID NO: 82), and a CDR3 comprising an amino acid sequence of AANGWGLDPTTYHY (SEQ ID NO: 83);xviii). a CDR1 comprising an amino acid sequence of GFTFGALA (SEQ ID NO: 86), a CDR2 comprising an amino acid sequence of ISNDGEHI (SEQ ID NO: 87), and a CDR3 comprising an amino acid sequence of SAGWTRRIFQY (SEQ ID NO: 88);xix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92);14323195900v2Attorney Docket No: 260525.000128xx). a CDR1 comprising an amino acid sequence of GFTLSGYA (SEQ ID NO: 95), a CDR2 comprising an amino acid sequence of ITSAGGST (SEQ ID NO: 96), and a CDR3 comprising an amino acid sequence of KAGIRGEVY (SEQ ID NO: 97);xxi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);xxii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);xxiii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);xxiv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of GAASSWSRGGVPYGMDY (SEQ ID NO: 2298);xxv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AGASSWSRGGVPYGMDY (SEQ ID NO: 2299);xxvi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAGSSWSRGGVPYGMDY (SEQ ID NO: 2300);xxvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAAASWSRGGVPYGMDY (SEQ ID NO: 2301);xxviii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASAWSRGGVPYGMDY (SEQ ID NO: 2302);xxix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSASRGGVPYGMDY (SEQ ID NO: 2303);15323195900v2Attorney Docket No: 260525.000128xxx). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWARGGVPYGMDY (SEQ ID NO: 2304);xxxi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSAGGVPYGMDY (SEQ ID NO: 2305);xxxii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRAGVPYGMDY (SEQ ID NO: 2306);xxxiii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGAVPYGMDY (SEQ ID NO: 2307);xxxiv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGAPYGMDY (SEQ ID NO: 2308);xxxv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVAYGMDY (SEQ ID NO: 2309);xxxvi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPAGMDY (SEQ ID NO: 2310);xxxvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYAMDY (SEQ ID NO: 2311);xxxviii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGADY (SEQ ID NO: 2312);xxxix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMAY (SEQ ID NO: 2313);16323195900v2Attorney Docket No: 260525.000128xl). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDA (SEQ ID NO: 2314);xli). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of AAGPTMSYSRGGEF (SEQ ID NO: 2315);xlii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TGGPTMSYSRGGEF (SEQ ID NO: 2316);xliii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAAPTMSYSRGGEF (SEQ ID NO: 2317);xliv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGATMSYSRGGEF (SEQ ID NO: 2318);xlv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPAMSYSRGGEF (SEQ ID NO: 2319);xlvi). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTASYSRGGEF (SEQ ID NO: 2320);xlvii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMAYSRGGEF (SEQ ID NO: 2321);xlviii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSASRGGEF (SEQ ID NO: 2322);xlix). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYARGGEF (SEQ ID NO: 2323);17323195900v2Attorney Docket No: 260525.000128I), a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSAGGEF (SEQ ID NO: 2324);li). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRAGEF (SEQ ID NO: 2325);lii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGAEF (SEQ ID NO: 2326);liii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGAF (SEQ ID NO: 2327);liv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEA (SEQ ID NO: 2328);Iv). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of AAYNDGGEY (SEQ ID NO: 2329);Ivi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RGYNDGGEY (SEQ ID NO: 2330);Ivii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAANDGGEY (SEQ ID NO: 2331);Iviii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYADGGEY (SEQ ID NO: 2332);lix). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNAGGEY (SEQ ID NO: 2333);18323195900v2Attorney Docket No: 260525.000128lx), a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDAGEY (SEQ ID NO: 2334);Ixi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGAEY (SEQ ID NO: 2335);Ixii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGAY (SEQ ID NO: 2336);Ixiii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEA (SEQ ID NO: 2337);Ixiv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of ASRWYNL (SEQ ID NO: 2338);Ixv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HARWYNL (SEQ ID NO: 2339);Ixvi). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSAWYNL (SEQ ID NO: 2340);Ixvii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRAYNL (SEQ ID NO: 2341);Ixviii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWANL (SEQ ID NO: 2342);Ixix). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYAL (SEQ ID NO: 2343);19323195900v2Attorney Docket No: 260525.000128Ixx). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNA (SEQ ID NO: 2344);Ixxi). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of ATADY (SEQ ID NO: 2345);Ixxii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FAADY (SEQ ID NO: 2346);Ixxiii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTGDY (SEQ ID NO: 2347); orIxxiv). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTAAY (SEQ ID NO: 2348).

[0055] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise: i). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), and a CDR3 comprising an amino acid sequence of NAARSYSRDYEPLKPDY (SEQ ID NO: 13);ii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRAGEF (SEQ ID NO: 2325);iii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYARGGEF (SEQ ID NO: 2323);iv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);v). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);20323195900v2Attorney Docket No: 260525.000128vi). a CDR1 comprising an amino acid sequence of GFTLSGYA (SEQ ID NO: 95), a CDR2 comprising an amino acid sequence of ITSAGGST (SEQ ID NO: 96), and a CDR3 comprising an amino acid sequence of KAGIRGEVY (SEQ ID NO: 97);vii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);viii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);ix). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);x). a CDR1 comprising an amino acid sequence of GLPFRSYA (SEQ ID NO: 2260), a CDR2 comprising an amino acid sequence of ISSGGGIV (SEQ ID NO: 2271), and a CDR3 comprising an amino acid sequence of FASDY (SEQ ID NO: 2276);xi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of MKWTGNT (SEQ ID NO: 73), and a CDR3 comprising an amino acid sequence of TAGPAISLSRGGEY (SEQ ID NO: 74);xii). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orxiii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92).

[0056] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise: i). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);ii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);21323195900v2Attorney Docket No: 260525.000128iii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);iv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);v). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);vi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92).

[0057] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise at least one anti-TRAILR2 single-domain antibody.

[0058] In some embodiments, the anti-TRAILR2 single-domain antibody is an anti-TRAILR2 VHH, an anti-TRAILR2 VNAR, or an anti-TRAILR2 VH domain.

[0059] In some embodiments, the anti-TRAILR2 single-domain antibody is an anti-TRAILR2 VHH.

[0060] In some embodiments, the anti-TRAILR2 VHH is a camelid anti-TRAILR2 VHH.

[0061] In some embodiments, the camelid anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 4, 9, 14, 17, 21, 26, 31, 36, 41, 45, 50, 55, 65, 70, 75, 79, 84, 89, 93, 98, 146-468, 2192, 2198, 2204, and 2214-2233, or an amino acid sequence having at least 75% identity thereto.

[0062] In some embodiments, the camelid anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 14, 31, 55, 2192, 2198, and 2204, or an amino acid sequence having at least 75% identity thereto.

[0063] In some embodiments, the anti-TRAILR2 VHH is a humanized anti-TRAILR2 VHH.

[0064] In some embodiments, the humanized anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 5, 10, 15, 18, 22, 27, 32, 37, 42, 46, 51, 56, 66, 71, 76, 80, 85, 90,22323195900v2Attorney Docket No: 260525.00012894, 99, 469-761, 2194, 2200, 2206, 2234-2251, 2354-2404, 4662-4677, and 4692-4697, or an amino acid sequence having at least 75% identity thereto.

[0065] In some embodiments, the humanized anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 32, 56, 76, 80, 94, 99, 2194, 2200, 2206, 2250, 2379, 2381, 4662-4677, and 4692-4697, or an amino acid sequence having at least 75% identity thereto

[0066] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains that specifically binds to human TRAILR2. In some embodiments, the one or more anti-TRAILR2 antigenbinding domains bind to human TRAILR2 with a KDof less than about 3xl0“7M. In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind to human TRAILR2 with a KDof about 1×10-10to 5×10-8M.

[0067] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind to cyno TRAILR2. In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind to cyno TRAILR2 with a KDof less than about 3×10-7M. In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind to cyno TRAILR2 with a KDof about lxlO"9to lxlO"7M.

[0068] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains do not block binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2.

[0069] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains block binding of TRAILto TRAILR2.

[0070] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains do not bind specifically to TRAILR1, TRAILR3 and / or TRAILR4.

[0071] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind cysteine-rich domain (CRD) domain 1 (CRD1) and / or CRD2 of TRAILR2.

[0072] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind CRD1 of TRAILR2.

[0073] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind CRD2 of TRAILR2.

[0074] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind CRD1 and CRD2 ofTRAILR2.

[0075] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind CRD2 and CRD3 ofTRAILR2.

[0076] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind to the same epitope on TRAILR2.23323195900v2Attorney Docket No: 260525.000128

[0077] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains bind to different epitopes on TRAILR2.

[0078] In various embodiments, the one or more anti-CDH17 antigen-binding domains comprise a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from:a) A(A / L)CLLRFE(S / T)CLEYNRAQY(N / P)Y (SEQ ID NO: 2583);b) AAVR(A / S)GSDWWTTM(R / T)QR(D / H)YD(F / Y) (SEQ ID NO: 2585);c) AARDSR(K / R)GGLFADLN(E / G)YDY (SEQ ID NO: 2588);d) AATG(D / N)(L / S)YRGAYDRP(A / T)EYDY (SEQ ID NO: 2590);e) VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529);f) AAQFSLPVDA(S / T)PLRRY(A / Y)(H / Y) (SEQ ID NO: 2593);g) AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);h) AARNGGYDLNDYAY (SEQ ID NO: 2544);i) NAGG(G / P / A)(A / R)(L / R)GY (SEQ ID NO: 2598);j) NVGGQL(K / L / R)GY (SEQ ID NO: 2601);k) NQGG(Q / S)KGY (SEQ ID NO: 2604);l) AADG(L / P)PY(G / S)(D / S)WFGDQFDV (SEQ ID NO: 4133);m) AFNKWGRLSADL(D / N)DYFR (SEQ ID NO: 4136);n) N(M / T)HRSY(A / D)I(D / N / R / S)FYDN (SEQ ID NO: 4139); ando) RRYDDY(D / G)S (SEQ ID NO: 4142),wherein one or more non-alanine residues in the CDR3 sequence are optionally replaced with an alanine, and / or one or more alanine residues in the CDR3 sequence are optionally replaced with a glycine.

[0079] In some embodiments, the CDR3 of the one or more anti-CDH17 antigen-binding domains comprises an amino acid sequence selected from: ALCLLRFETCLEYNRAQYPY (SEQ ID NO: 2503);AAVRSGSDWWTTMTQRHYDF (SEQ ID NO: 2508); AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513);AATGDSYRGAYDRPAEYDY (SEQ ID NO: 2518); AATGNSYRGAYDRPTEYDY (SEQ ID NO: 2522);AATGDLYRGAYDRPAEYDY (SEQ ID NO: 2525); VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529);AAQFSLPVDASPLRRYYY (SEQ ID NO: 2534); AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);AARNGGYDLNDYAY (SEQ ID NO: 2544); NAGGGRLGY (SEQ ID NO: 2549); NAGGAALGY (SEQ ID NO: 2554); NVGGQLLGY (SEQ ID NO: 2559); NQGGSKGY (SEQ ID NO: 2564); AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613); AADGLPYGDWFGDQFDV (SEQ ID NO: 4109); AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);NMHRSYDISFYDN (SEQ ID NO: 4115); and RRYDDYGS (SEQ ID NO: 4118).24323195900v2Attorney Docket No: 260525.000128

[0080] In some embodiments, the one or more anti-CDH17 antigen-binding domains further comprise a CDR1 comprising an amino acid sequence selected from:a). GFTLSN(T / Y)N (SEQ ID NO: 2581);b). GSPLDYYA (SEQ ID NO: 2506);c). RL(A / N / T)(F / S)(N / S)(R / S)(S / T)T;d). GRTFS(E / T)PI (SEQ ID NO: 2589);e). GRTFSSPI (SEQ ID NO: 2528);f). TRTF(D / N)MYA (SEQ ID NO: 2591);g). GRTF(D / S)S(L / Y)(L / V) (SEQ ID NO: 2594);h). GRTDSILN (SEQ ID NO: 2542);i). G(I / M)RFS(S / Q)YA (SEQ ID NO: 2596);j). GIRFS(A / S)YA (SEQ ID NO: 2599);k). GS(I / R)FS(R / S)(W / Y)A (SEQ ID NO: 2602);l). G(G / R)TASEYG (SEQ ID NO: 4131);m). EQTMTGF(T / W) (SEQ ID NO: 4134);n). GLRFS(S / N)YA (SEQ ID NO: 4137); ando). (G / R)GT(F / V)SGYA (SEQ ID NO: 4140).

[0081] In some embodiments, the CDR1 of the one or more anti-CDH17 antigen-binding domains comprises an amino acid sequence selected from GFTLSNYN (SEQ ID NO: 2501); GSPLDYYA (SEQ ID NO: 2506); RLNFSRTT (SEQ ID NO: 2511); GRTFSEPI (SEQ ID NO: 2516); GRTFSTPI (SEQ ID NO: 2521);GRTFSEPI (SEQ ID NO: 2516); GRTFSSPI (SEQ ID NO: 2528); TRTFNMYA (SEQ ID NO: 2532); GRTFSSYL (SEQ ID NO: 2537); GRTDSILN (SEQ ID NO: 2542); GIRFSSYA (SEQ ID NO: 2547); GMRFSQYA (SEQ ID NO: 2552); GIRFSAYA (SEQ ID NO: 2557); GSRFSSYA (SEQ ID NO: 2562); TRTFDMYA (SEQ ID NO: 3137);GGTASEYG (SEQ ID NO: 4107); EQTMTGFW (SEQ ID NO: 4110); GLRFSNYA (SEQ ID NO: 4113); and GGTVSGYA (SEQ ID NO: 4116).

[0082] In some embodiments, the one or more anti-CDH17 antigen-binding domains further comprise a CDR2 comprising an amino acid sequence selected from:a) (F / I)SRGGRT (SEQ ID NO: 2582);b) ISTSGR(C / S)T (SEQ ID NO: 2584);c) SGW(A / S)R(G / T)RT (SEQ ID NO: 2587);d) LISTGGST (SEQ ID NO: 2517);e) l(N / S)RSG(A / T)NT (SEQ ID NO: 2592);25323195900v2Attorney Docket No: 260525.000128f) ISWN(D / G)RST (SEQ ID NO: 2595);g) ISWFRGET (SEQ ID NO: 2543);h) l(F / T)(l / K / N / S)(D / G)(G / Y)(R / S / T)T;i) MT(A / N / T)GGMT (SEQ ID NO: 2600);j) IT(N / S)GG(G / R / S)T (SEQ ID NO: 2603);k) ISTSGGVT (SEQ ID NO: 4108);l) ISASGSRV (SEQ ID NO: 4111);m) IT(N / K)GG(I / N / S)T (SEQ ID NO: 4138); andn) INSGGPT (SEQ ID NO: 4117).

[0083] In some embodiments, the CDR2 of the one or more anti-CDH17 antigen-binding domains comprises an amino acid sequence selected ISRGGRT (SEQ ID NO: 2502); ISTSGRCT (SEQ ID NO: 2507); SGWARGRT (SEQ ID NO: 2512); LISTGGST (SEQ ID NO: 2517); ISRSGTNT (SEQ ID NO: 2533); ISWNDRST (SEQ ID NO: 2538); ISWFRGET (SEQ ID NO: 2543); ITSGYRT (SEQ ID NO: 2548); IFKDGTT (SEQ ID NO: 2553); MTAGGMT (SEQ ID NO: 2558); ITSGGRT (SEQ ID NO: 2563); INRSGANT (SEQ ID NO: 3375);ISTSGGVT (SEQ ID NO: 4108); ISASGSRV (SEQ ID NO: 4111); ITKGGIT (SEQ ID NO: 4114); and INSGGPT (SEQ ID NO: 4117).

[0084] In some embodiments, the one or more anti-CDH17 antigen-binding domains comprise a CDR1 comprising an amino acid sequence selected from SEQ ID Nos: 2501, 2506, 2511, 2516, 2521, 2528, 2532, 2537, 2542, 2547, 2552, 2557, 2562, 3055-3292, 4107, 4110, 4113, 4116, 4192-4220; a CDR2 comprising an amino acid sequence selected from SEQ ID Nos: 2502, 2507, 2512, 2517, 2533, 2538, 2543, 2548, 2553, 2558, 2563, 3293-3530, 4108, 4111, 4114, 4117, 4221-4248; and / or a CDR3 comprising an amino acid sequence selected from SEQ ID Nos: 2503, 2508, 2513, 2518, 2522, 2525, 2529, 2534, 2539, 2544, 2549, 2554, 2559, 2564, 3531-3768, 4109, 4112, 4115, 4118, 4249-4277.

[0085] In some embodiments, the one or more anti-CDH17 antigen-binding domains comprise: i). a CDR1 comprising an amino acid sequence of GFTLSNYN (SEQ ID NO: 2501), a CDR2 comprising an amino acid sequence of ISRGGRT (SEQ ID NO: 2502), and a CDR3 comprising an amino acid sequence of ALCLLRFETCLEYNRAQYPY (SEQ ID NO: 2503);ii). a CDR1 comprising an amino acid sequence of GSPLDYYA (SEQ ID NO: 2506), a CDR2 comprising an amino acid sequence of ISTSGRCT (SEQ ID NO: 2507), and a CDR3 comprising an amino acid sequence of AAVRSGSDWWTTMTQRHYDF (SEQ ID NO: 2508);26323195900v2Attorney Docket No: 260525.000128iii). a CDR1 comprising an amino acid sequence of RLNFSRTT (SEQ ID NO: 2511), a CDR2 comprising an amino acid sequence of SGWARGRT (SEQ ID NO: 2512), and a CDR3 comprising an amino acid sequence of AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513);iv). a CDR1 comprising an amino acid sequence of GRTFSEPI (SEQ ID NO: 2516), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of AATGDSYRGAYDRPAEYDY (SEQ ID NO: 2518);v). a CDR1 comprising an amino acid sequence of GRTFSTPI (SEQ ID NO: 2521), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of AATGNSYRGAYDRPTEYDY (SEQ ID NO: 2522);vi). a CDR1 comprising an amino acid sequence of GRTFSEPI (SEQ ID NO: 2516), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of AATGDLYRGAYDRPAEYDY (SEQ ID NO: 2525);vii). a CDR1 comprising an amino acid sequence of GRTFSSPI (SEQ ID NO: 2528), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529);viii). a CDR1 comprising an amino acid sequence of TRTFNMYA (SEQ ID NO: 2532), a CDR2 comprising an amino acid sequence of ISRSGTNT (SEQ ID NO: 2533), and a CDR3 comprising an amino acid sequence of AAQFSLPVDASPLRRYYY (SEQ ID NO: 2534);ix). a CDR1 comprising an amino acid sequence of GRTFSSYL (SEQ ID NO: 2537), a CDR2 comprising an amino acid sequence of ISWNDRST (SEQ ID NO: 2538), and a CDR3 comprising an amino acid sequence of AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);x). a CDR1 comprising an amino acid sequence of GRTDSILN (SEQ ID NO: 2542), a CDR2 comprising an amino acid sequence of ISWFRGET (SEQ ID NO: 2543), and a CDR3 comprising an amino acid sequence of AARNGGYDLNDYAY (SEQ ID NO: 2544);xi). a CDR1 comprising an amino acid sequence of GIRFSSYA (SEQ ID NO: 2547), a CDR2 comprising an amino acid sequence of ITSGYRT (SEQ ID NO: 2548), and a CDR3 comprising an amino acid sequence of NAGGGRLGY (SEQ ID NO: 2549);xii). a CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554);27323195900v2Attorney Docket No: 260525.000128xiii). a CDR1 comprising an amino acid sequence of GIRFSAYA (SEQ ID NO: 2557), a CDR2 comprising an amino acid sequence of MTAGGMT (SEQ ID NO: 2558), and a CDR3 comprising an amino acid sequence of NVGGQLLGY (SEQ ID NO: 2559);xiv). a CDR1 comprising an amino acid sequence of GSRFSSYA (SEQ ID NO: 2562), a CDR2 comprising an amino acid sequence of ITSGGRT (SEQ ID NO: 2563), and a CDR3 comprising an amino acid sequence of NQGGSKGY (SEQ ID NO: 2564);xv). a CDR1 comprising an amino acid sequence of TRTFDMYA (SEQ ID NO: 3137), a CDR2 comprising an amino acid sequence of INRSGANT (SEQ ID NO: 3375), and a CDR3 comprising an amino acid sequence of AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613);xvi). a CDR1 comprising an amino acid sequence of GGTASEYG (SEQ ID NO: 4107), a CDR2 comprising an amino acid sequence of ISTSGGVT (SEQ ID NO: 4108), and a CDR3 comprising an amino acid sequence of AADGLPYGDWFGDQFDV (SEQ ID NO: 4109);xvii) a CDR1 comprising an amino acid sequence of EQTMTGFW (SEQ ID NO: 4110), a CDR2 comprising an amino acid sequence of ISASGSRV (SEQ ID NO: 4111), and a CDR3 comprising an amino acid sequence of AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);xviii) a CDR1 comprising an amino acid sequence of GLRFSNYA (SEQ ID NO: 4113), a CDR2 comprising an amino acid sequence of ITKGGIT (SEQ ID NO: 4114), and a CDR3 comprising an amino acid sequence of NMHRSYDISFYDN (SEQ ID NO: 4115); orxix). a CDR1 comprising an amino acid sequence of GGTVSGYA (SEQ ID NO: 4116), a CDR2 comprising an amino acid sequence of INSGGPT (SEQ ID NO: 4117), and a CDR3 comprising an amino acid sequence of RRYDDYGS (SEQ ID NO: 4118).

[0086] In some embodiments, the one or more anti-CDH17 antigen-binding domains comprise: i). a CDR1 comprising an amino acid sequence of GRTDSILN (SEQ ID NO: 2542), a CDR2 comprising an amino acid sequence of ISWFRGET (SEQ ID NO: 2543), and a CDR3 comprising an amino acid sequence of AARNGGYDLNDYAY (SEQ ID NO: 2544);ii). a CDR1 comprising an amino acid sequence of GIRFSSYA (SEQ ID NO: 2547), a CDR2 comprising an amino acid sequence of ITSGYRT (SEQ ID NO: 2548), and a CDR3 comprising an amino acid sequence of NAGGGRLGY (SEQ ID NO: 2549);iii). a CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554);28323195900v2Attorney Docket No: 260525.000128iv). a CDR1 comprising an amino acid sequence of RLNFSRTT (SEQ ID NO: 2511), a CDR2 comprising an amino acid sequence of SGWARGRT (SEQ ID NO: 2512), and a CDR3 comprising an amino acid sequence of AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513);v). a CDR1 comprising an amino acid sequence of TRTFDMYA (SEQ ID NO: 3137), a CDR2 comprising an amino acid sequence of INRSGANT (SEQ ID NO: 3375), and a CDR3 comprising an amino acid sequence of AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613);vi). a CDR1 comprising an amino acid sequence of GRTFSSYL (SEQ ID NO: 2537), a CDR2 comprising an amino acid sequence of ISWNDRST (SEQ ID NO: 2538), and a CDR3 comprising an amino acid sequence of AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);vii). a CDR1 comprising an amino acid sequence of GGTASEYG (SEQ ID NO: 4107), a CDR2 comprising an amino acid sequence of ISTSGGVT (SEQ ID NO: 4108), and a CDR3 comprising an amino acid sequence of AADGLPYGDWFGDQFDV (SEQ ID NO: 4109);viii). a CDR1 comprising an amino acid sequence of EQTMTGFW (SEQ ID NO: 4110), a CDR2 comprising an amino acid sequence of ISASGSRV (SEQ ID NO: 4111), and a CDR3 comprising an amino acid sequence of AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);ix). a CDR1 comprising an amino acid sequence of GLRFSNYA (SEQ ID NO: 4113), a CDR2 comprising an amino acid sequence of ITKGGIT (SEQ ID NO: 4114), and a CDR3 comprising an amino acid sequence of NMHRSYDISFYDN (SEQ ID NO: 4115); orx). a CDR1 comprising an amino acid sequence of GGTVSGYA (SEQ ID NO: 4116), a CDR2 comprising an amino acid sequence of INSGGPT (SEQ ID NO: 4117), and a CDR3 comprising an amino acid sequence of RRYDDYGS (SEQ ID NO: 4118).

[0087] In some embodiments, the one or more anti-CDH17 antigen-binding domains comprise: a), a CDR1 comprising an amino acid sequence of GGTASEYG (SEQ ID NO: 4107), a CDR2 comprising an amino acid sequence of ISTSGGVT (SEQ ID NO: 4108), and a CDR3 comprising an amino acid sequence of AADGLPYGDWFGDQFDV (SEQ ID NO: 4109);b). a CDR1 comprising an amino acid sequence of EQTMTGFW (SEQ ID NO: 4110), a CDR2 comprising an amino acid sequence of ISASGSRV (SEQ ID NO: 4111), and a CDR3 comprising an amino acid sequence of AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);c). a CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554); or29323195900v2Attorney Docket No: 260525.000128d). a CDR1 comprising an amino acid sequence of GGTVSGYA (SEQ ID NO: 4116), a CDR2 comprising an amino acid sequence of INSGGPT (SEQ ID NO: 4117), and a CDR3 comprising an amino acid sequence of RRYDDYGS (SEQ ID NO: 4118).

[0088] In some embodiments, the one or more anti-CDH17 antigen-binding domains comprise at least one anti-CDH17 single-domain antibody.

[0089] In some embodiments, the anti-CDH17 single-domain antibody is an anti-CDH17 VHH, an anti-CDH17 VNAR, or an anti-CDH17 VH domain.

[0090] In some embodiments, the anti-CDH17 single-domain antibody is an anti-CDH17 VHH.

[0091] In some embodiments, the anti-CDH17 VHH is a camelid anti-CDH17 VHH.

[0092] In some embodiments, the camelid anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2504, 2509, 2514, 2519, 2523, 2526, 2530, 2535, 2540, 2545, 2550, 2555, 2560, 2565, 2605-2842, 4119-4122, 4143-4171, and 4278-4306, or an amino acid sequence having at least 75% identity thereto.

[0093] In some embodiments, the camelid anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2545, 2555, 4119, 4120, and 4122, or an amino acid sequence having at least 75% identity thereto.

[0094] In some embodiments, the anti-CDH17 VHH is a humanized anti-CDH17 VHH.

[0095] In some embodiments, the humanized anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2505, 2510, 2515, 2520, 2524, 2527, 2531, 2536, 2541, 2546, 2551, 2556, 2561, 2566, 2843-3054, 4123-4126, 4172-4191, 4651-4659, and 4691, or a sequence having at least 75% identity thereto.

[0096] In some embodiments, the humanized anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2515, 2541, 2556, 4123, 4124, 4126, 4651-4659, and 4691, or a sequence having at least 75% identity thereto.

[0097] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to human CDH17. In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to human CDH17 with a KDof about 2.2xl0“7M or less. In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to human CDH17 with a KDof about 1.5xl0“7M or less.

[0098] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to cyno CDH17. In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to cyno CDH17 with a KDof about 2xl0“8M or less.30323195900v2Attorney Docket No: 260525.000128

[0099] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to mouse CDH17.

[0100] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to the extracellular cadherin domain 6 (EC6) and / or extracellular cadherin domain 7 (EC7) of human CDH17.

[0101] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to an extracellular region C-terminal to the tripeptide motif, RGD, in EC6 of human CDH17.

[0102] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to the extracellular cadherin domain 1 (ECI) of human CDH17.

[0103] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to the same epitope on CDH17.

[0104] In some embodiments, the one or more anti-CDH17 antigen-binding domains bind to different epitopes on CDH17.

[0105] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise i). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);ii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);iii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);iv). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92); andthe one or more anti-CDH17 antigen-binding domains comprisea CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554).31323195900v2Attorney Docket No: 260525.000128

[0106] In some embodiments, the antigen binding domain(s) that specifically bind to HSA comprise:i). a CDR1 comprising an amino acid sequence of GFTFTDYS (SEQ ID NO: 4678), a CDR2 comprising an amino acid sequence of TRTTGGSI (SEQ ID NO: 4679), and a CDR3 comprising an amino acid sequence of LAYRQKGSNSPLIV (SEQ ID NO: 4680); orii). a CDR1 comprising an amino acid sequence of GFTFTDYS (SEQ ID NO: 4678), a CDR2 comprising an amino acid sequence of TRTTGGSI (SEQ ID NO: 4679), and a CDR3 comprising an amino acid sequence of LAYRQKGSNSPLII (SEQ ID NO: 4681).

[0107] In some embodiments, the one or more anti-HSA antigen-binding domains comprise at least one anti-HSA single-domain antibody.

[0108] In some embodiments, the anti-HSA single-domain antibody is an anti-HSA VHH, an anti-HSA VNAR, or an anti-HSA VH domain.

[0109] In some embodiments, the anti-HSA single-domain antibody is an anti-HSA VHH.

[0110] In some embodiments, the anti-HSA VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 4660 and 4661, or an amino acid sequence having at least 75% identity thereto.

[0111] In some embodiments, the antigen-binding protein comprises one or more modifications that reduce binding of said antigen-binding protein by pre-existing antibodies found in human blood or serum.

[0112] In some embodiments, the one or more antigen-binding domains are one or more VHHs, and at least one VHH comprises one or more modifications at the amino-terminus and / or the carboxyterminus.

[0113] In some embodiments, the one or more antigen-binding domains are one or more VHHs, and wherein the amino acid sequence at the carboxy-terminus of at least one VHH starting from position 111 according to Chothia comprises VAGG (SEQ ID NO: 2405) or VPAG (SEQ ID NO: 2406).

[0114] In some embodiments, the one or more antigen-binding domains are one or more VHHs, and wherein the amino acid residue Glu at the first position of at least one VHH is replaced with Asp (EID).

[0115] In some embodiments, the immunoglobulin Fc region is an Fc region of a human immunoglobulin.

[0116] In some embodiments, the immunoglobulin Fc region is an Fc region of human IgGl, lgG2, lgG3 or lgG4, or a variant thereof.

[0117] In some embodiments, the immunoglobulin Fc region is an Fc region of human IgGl, or a variant thereof.32323195900v2Attorney Docket No: 260525.000128

[0118] In some embodiments, the Fc region of human IgGl comprises one or more mutations selected from Leu234Ala (L234A), Leu234Gly (L234G), Leu234Ser (L234S), Leu234Thr (L234T), Leu235Ala (L235A), Leu235Glu (L235E), Leu235Ser (L235S), Leu235Thr (L235T), Leu235Val (L235V), Leu235Gln (L235Q), Gly236Arg (G236R), Met252Tyr (M252Y), Ser254Thr (S254T), Thr256Glu (T256E), Asp265Asn (D265N), Asp265Ala (D265A), Asp270Asn (D270N), Ser298Asn (S298N), Asn297Ala (N297A), Pro329Ala (P329A), Pro239Gly (P329G), Asn325Glu (N325E) and / or Ala327Ser (A327S) according to EU numbering.

[0119] In some embodiments, the Fc region of human IgGl comprises a set of mutations selected from1). L234A and L235A;2). L234A, L235A, and P329A;3). D265A, N297A and P329A;4). L234A, L235A, and G237A;5). L234G, L235S, and G236R;6). L234S, L235T, and G236R;7). L234S, L235V, and G236R;8). L234T, L235Q, and G236R;9). L234T, L235T, and G236R;10). L234A, L235A, and P329G; and11). M252Y, S254T, and T256E.

[0120] In some embodiments, the immunoglobulin Fc region is an Fc region of human lgG4, or a variant thereof.

[0121] In some embodiments, the Fc region of human lgG4 comprises one or more mutations selected from Ser228Pro (S228P), Leu235Glu (L235E), Leu235Ala (L235A), Phe234Ala (F234A), and / or Pro329Gly (P329G) according to EU numbering.

[0122] In some embodiments, the Fc region of human lgG4 comprises a set of mutations selected from1). S228P and L235E;2). S228P and L235A;3). S228P, F234A, and L235E;4). S228P, F234A, and L235A; and5) P329G, S228P, and L235E.33323195900v2Attorney Docket No: 260525.000128

[0123] In some embodiments, the Fc region used in a multispecific antigen-binding protein of the present disclosure comprises an amino acid sequence selected from:DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 2152);DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNS TYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO: 2153);ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 2091); or ESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQF NSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 2156).

[0124] In some embodiments, the multispecific antigen-binding protein comprises at least one linker, wherein said linker is selected from the group consisting of SEQ ID NOs: 2088-2090, 2092-2125, 2132-2148, 2180, 2408, 4007-4041, 4048-4067, and 4683.

[0125] In some embodiments, the at least one linker is a rigid linker. In some embodiments, the at least one rigid linker is selected from the group consisting of GGGGSPAPAPAPAPAPAPAPAPGGGS (SEQ ID NO: 2088); PAPAPAPAPAPAPAPAP (SEQ ID NO: 2132), GGGGSPAPAPAPAPAPAPAPAPGGGGS (SEQ ID NO: 2135), GGGGSPAPAPAPAPGGGGS (SEQ ID NO: 4698), GGGGSPAPAPAPAPAPAPAPAPAPAPAPAPGGGGS (SEQ ID NO: 4699), and A(EAAAK)nA (SEQ ID NO: 2148), where n is any integer, e.g., 1, 2, 3, 4, 5, 6, or 7, 8, 9 or 10.

[0126] In some embodiments, the at least one linker is a flexible linker. In some embodiments, the at least one flexible linker is selected from the group consisting of GGGGSGGGS (SEQ ID NO: 4683); Gn(SEQ ID NO: 4700, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10); GnS (SEQ ID NO: 2136, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10), SGn(SEQ ID NO: 2137, where n is any integer, e.g., 1, 2, 3,34323195900v2Attorney Docket No: 260525.0001284, 5, 6, or 7, 8, 9 or 10), and (GGGGS)n(SEQ ID NO: 2138, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, or 7, 8, 9 or 10).

[0127] In some embodiments, the multispecific antigen-binding protein comprises the amino acid sequence of any one of SEQ ID NOs: 4500-4506, 4509-4524, and 4526-4650, or a sequence having at least 70% identity thereto.

[0128] In some embodiments, the multispecific antigen-binding protein comprises a single polypeptide chain, said single polypeptide chain comprising an amino acid sequence selected from any one of SEQ ID NOs: 4500-4506, 4509-4524, and 4526-4584, or a sequence having at least 70% identity thereto.

[0129] In some embodiments, the multispecific antigen-binding protein comprises a first polypeptide chain and a second polypeptide chain, each polypeptide chain comprising an amino acid sequence selected from any one of SEQ ID NOs: 4585-4650, or a sequence having at least 70% identity thereto. In some embodiments, each polypeptide chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 4587-4589, 4603, 4625, 4626, 4633, 4641, 4643, and 4644, or a sequence having at least 70% identity thereto. In one embodiment, each polypeptide chain comprises an amino acid sequence of SEQ ID NO: 4626, or a sequence having at least 70% identity thereto.

[0130] In another aspect, provided herein is a conjugate comprising the multispecific antigen-binding protein of the present disclosure, wherein the multispecific antigen-binding protein is conjugated to a second moiety. In some embodiments, the second moiety is selected from a detectable label, a drug, a toxin, a radionuclide, an enzyme, an immunomodulatory agent, a cytokine, a cytotoxic agent, a chemotherapeutic agent, and a diagnostic agent, or a combination thereof.

[0131] In another aspect, provided herein is a polynucleotide molecule encoding the multispecific antigen-binding protein of the present disclosure.

[0132] In some embodiments, the polynucleotide molecule comprises the nucleotide sequence of any one of SEQ ID NOs: 57-61, 131-145, 1731-2053, 2071-2087, 2193, 2199, 2205, 2278-2297, 2567-2580, 3769-4006, 4127-4130, and 4307-4335 or a nucleotide sequence having at least 70% identity thereto.

[0133] In some embodiments, the polynucleotide molecule comprises the nucleotide sequence of any one of SEQ ID NOs: 57-61, 131-145, 1731-2053, 2071-2087, 2193, 2199, 2205, and 2278-2297, or a nucleotide sequence having at least 70% identity thereto.35323195900v2Attorney Docket No: 260525.000128

[0134] In some embodiments, the polynucleotide molecule comprises the nucleotide sequence of any one of SEQ ID NOs: 57-61, 131-145, 2071-2087, 2193, 2199, and 2205, or a nucleotide sequence having at least 70% identity thereto.

[0135] In some embodiments, the polynucleotide molecule comprises the nucleotide sequence of any one of SEQ ID NOs: 132, 137, 2193, 2199, and 2205, or a nucleotide sequence having at least 70% identity thereto.

[0136] In some embodiments, the polynucleotide molecule comprises the nucleotide sequence of any one of SEQ ID NOs: 2567-2580, 3769-4006, 4127-4130, and 4307-4335, or a nucleotide sequence having at least 70% identity thereto.

[0137] In some embodiments, the polynucleotide molecule comprises the nucleotide sequence of any one of SEQ ID NOs: 2567-2580, 3851, and 4127-4130, or a nucleotide sequence having at least 70% identity thereto.

[0138] In another aspect, provided herein is a recombinant vector comprising the polynucleotide molecule of the present disclosure.

[0139] In another aspect, provided herein is a host cell comprising the polynucleotide molecule of the present disclosure, or the recombinant vector of the present disclosure.

[0140] In another aspect, provided herein is a kit comprising the multispecific antigen-binding protein of the present disclosure, the conjugate of the present disclosure, the polynucleotide molecule of the present disclosure, the recombinant vector of the present disclosure, or the host cell of the present disclosure, and, optionally, instructions and / or packaging for the same.

[0141] In another aspect, provided herein is a pharmaceutical composition comprising the multispecific antigen-binding protein of the present disclosure, the conjugate of the present disclosure, the polynucleotide molecule of the present disclosure, or the recombinant vector of the present disclosure, and a pharmaceutically acceptable carrier and / or excipient.

[0142] In another aspect, provided herein is a method for preparing a multispecific antigen-binding protein that specifically binds TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2) and cadherin 17 (CDH17), comprising the steps of:a), culturing the host cell described herein in a culture medium under conditions suitable for expression of the multispecific antigen-binding protein; andb). isolating the multispecific antigen-binding protein from the host cell and / or the culture medium.36323195900v2Attorney Docket No: 260525.000128

[0143] In another aspect, provided herein is a method for inducing cell death in a cell expressing TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2) and / or cadherin 17 (CDH17), said method comprising contacting the cell with an effective amount of the antigen-binding protein or the conjugate of the present disclosure. In some embodiments, said contacting occurs in vitro. In some embodiments, said contacting occurs in vivo. In some embodiments, the cell expressing TRAILR2 and / or CDH17 is in a subject in need thereof. In some embodiments, the method further comprises administering the multispecific antigen-binding protein or the conjugate into the subject.

[0144] In some embodiments, the cell expressing TRAILR2 and / or CDH17 is a cell of a cancer. In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is selected from adenoid cystic carcinoma, adrenal gland tumor, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxiatelangiectasia, Beckwith-Wiedemann syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dube syndrome, bladder cancer, bone cancer (sarcoma of bone), brain stem glioma, brain tumor, breast cancer, inflammatory breast cancer, metastatic breast cancer, male breast cancer, Carney complex, central nervous system tumors (brain and spinal cord), cervical cancer, childhood cancer, colorectal cancer, Cowden syndrome, craniopharyngioma, desmoid tumor, desmoplastic infantile ganglioglioma, childhood tumor, ependymoma, esophageal cancer, Ewing sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial malignant melanoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell cancer, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal carcinoma, HIV / AIDS-related cancer, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumor, laryngeal and hypopharyngeal cancer, Li-Fraumeni syndrome, liver cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, lynch syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia type 1, multiple endocrine neoplasia type 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndromes (MDS), nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the lung, neuroendocrine tumor of the pancreas, neuroendocrine tumors, neurofibromatosis type 1, neurofibromatosis type 2, nevoid basal cell carcinoma syndrome, oral and oropharyngeal cancer, osteosarcoma, ovarian, fallopian tube, and peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers syndrome, pheochromocytoma and paraganglioma, pituitary gland tumor, pleuropulmonary blastoma, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer,37323195900v2Attorney Docket No: 260525.000128Kaposi sarcoma, soft tissue sarcomas, skin cancer (non-melanoma), small bowel cancer, stomach cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis complex, uterine cancer, vaginal cancer, Von Hippel-Lindau syndrome, vulvar cancer, Werner syndrome, Wilms tumor, and xeroderma pigmentosum.

[0145] In some embodiments, the cancer is selected from colorectal cancer, familial GIST, familial pancreatic cancer, gastrointestinal stromal tumor (GIST), hereditary diffuse gastric cancer, hereditary pancreatitis, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the pancreas, peritoneal cancer, pancreatic cancer, small bowel cancer, and stomach cancer.

[0146] In some embodiments, the cancer is gastrointestinal cancer, breast cancer, or lung cancer.

[0147] In some embodiments, the cancer is gastrointestinal cancer. In some embodiments, the gastrointestinal cancer is colorectal cancer, gastric cancer, esophageal cancer, pancreatic cancer, or cholangiocarcinoma cancer.

[0148] In some embodiments, the method further comprises contacting the cell with one or more additional therapeutic agents.

[0149] In another aspect, provided herein is a method of treating or preventing a cancer in a subject in need thereof, said method comprising administering to the subject an effective amount of the multispecific antigen-binding protein or the conjugate described herein.

[0150] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is selected from adenoid cystic carcinoma, adrenal gland tumor, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxia-telangiectasia, Beckwith-Wiedemann syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dube syndrome, bladder cancer, bone cancer (sarcoma of bone), brain stem glioma, brain tumor, breast cancer, inflammatory breast cancer, metastatic breast cancer, male breast cancer, Carney complex, central nervous system tumors (brain and spinal cord), cervical cancer, childhood cancer, colorectal cancer, Cowden syndrome, craniopharyngioma, desmoid tumor, desmoplastic infantile ganglioglioma, childhood tumor, ependymoma, esophageal cancer, Ewing sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial malignant melanoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell cancer, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal carcinoma, HIV / AIDS-related cancer, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumor, laryngeal and hypopharyngeal cancer, Li-Fraumeni syndrome, liver cancer, lung cancer, non-small cell lung cancer,38323195900v2Attorney Docket No: 260525.000128small cell lung cancer, lynch syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia type 1, multiple endocrine neoplasia type 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndromes (MDS), nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the lung, neuroendocrine tumor of the pancreas, neuroendocrine tumors, neurofibromatosis type 1, neurofibromatosis type 2, nevoid basal cell carcinoma syndrome, oral and oropharyngeal cancer, osteosarcoma, ovarian, fallopian tube, and peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers syndrome, pheochromocytoma and paraganglioma, pituitary gland tumor, pleuropulmonary blastoma, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, Kaposi sarcoma, soft tissue sarcomas, skin cancer (non-melanoma), small bowel cancer, stomach cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis complex, uterine cancer, vaginal cancer, Von Hippel-Lindau syndrome, vulvar cancer, Werner syndrome, Wilms tumor, and xeroderma pigmentosum.

[0151] In some embodiments, the cancer is selected from colorectal cancer, familial GIST, familial pancreatic cancer, gastrointestinal stromal tumor (GIST), hereditary diffuse gastric cancer, hereditary pancreatitis, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the pancreas, peritoneal cancer, pancreatic cancer, small bowel cancer, and stomach cancer.

[0152] In some embodiments, the cancer is gastrointestinal cancer, breast cancer, or lung cancer. In some embodiments, the cancer is gastrointestinal cancer.

[0153] In some embodiments, the gastrointestinal cancer is colorectal cancer, gastric cancer, esophageal cancer, pancreatic cancer, or cholangiocarcinoma cancer. In some embodiments, the method further comprises administering one or more additional therapeutic agents into the subject.

[0154] In some embodiments, the one or more additional therapeutic agents are selected from a chemotherapeutic agent, a vascular endothelial growth factor (VEGF) inhibitor, an epidermal growth factor receptor (EGFR) inhibitor, an apoptosis-inducing agent, and an immunotherapeutic agent, or a combination thereof.

[0155] In some embodiments, the chemotherapeutic agent is selected from bleomycin, carboplatin, chlorambucil, cisplatin, colchicine, cyclophosphamide, daunorubicin, doxorubicin or liposomal doxorubicin, mitomycin C, actinomycin, diethylstilbestrol, etoposide, 5-fluorouracil, floxuridine, melphalan, methotrexate, mitomycin, 6-mercaptopurine, teniposide, 6-thioguanine, vincristine and vinblastine, leflunomide, tamoxifen, interferon a-2b, glutamic acid, plicamycin, mercaptopurine, 6-39323195900v2Attorney Docket No: 260525.000128thioguanine, carmustine, BCNU, limousine, CCNU, cytosine arabinose, estramustine, hydroxyurea, procarbazine, busulfan, medroxyprogesterone, estramustine phosphate sodium, ethenyl estradiol, estradiol, megestrol acetate, methyltestosterone, diethylstilbestrol diphosphate, chlorotrianisene, testolactone, melphalan, chlorambucil, mechlorethamine, thiourea, bethamethasone sodium phosphate, dicarbazine, asparagine, mitotane, vincristine sulfate, vinblastine sulfate, FOLFOX (folinic acid, 5-fluorouracil and oxaliplatin), and FOLFIRI (folinic acid, 5-fluorouracil, and irinotecan), or a derivative or combination thereof. In one embodiment, the chemotherapeutic agent is an active metabolite of irinotecan, SN-38.

[0156] In some embodiments, the VEGF inhibitor is selected from bevacizumab, ramucirumab, regorafenib, and ziv-aflibercept, or a combination thereof.

[0157] In some embodiments, the EGFR inhibitor is selected from cetuximab and panitumumab, or a combination thereof.

[0158] In some embodiments, the apoptosis-inducing agent is selected from a B-cell lymphoma 2 (BCL2) inhibitor, a BCL-extra-large (BCL-XL) inhibitor, and an inhibitor of apoptosis proteins (IAP) inhibitor, or a combination thereof.

[0159] In some embodiments, the immunotherapeutic agent is selected from an anti-CTLA4 agent, an anti-PDl agent, an anti-PD-Ll agent, an anti-LAG3 agent, and an anti-TIM3 agent, or a combination thereof.

[0160] In some embodiments, the subject is a mammal. In some embodiments, the mammal is human.BRIEF DESCRIPTION OF THE DRAWINGS

[0161] Figure 1 depicts an exemplary general panning strategy for immune library construction and panning strategy for discovery of TRAILR2 binders.

[0162] Figure 2 shows sample selection for next generation sequencing (NGS) throughout the phage display process. For the three initial libraries, 12 TRAILR2 samples of the first panning round, and 18 TRAILR2 samples of the second panning round were sequenced with 20 million, 2 million, and 2 million reads, respectively. Comparison of V-body enrichment from the initial library to the first and second round of panning enabled the identification of potential V-body candidates.

[0163] Figure 3 shows a schematic diagram of an exemplary NGS workflow. Following phage display, the VHH region of the phage elutions was polymerase chain reaction (PCR) amplified, unique and sample-specific barcodes were fused, and NGS was performed with the Illumina NovaSeq platform from40323195900v2Attorney Docket No: 260525.000128Genewiz. The raw data were demultiplexed, and then processed by the Pipebio NGS analysis pipeline. Forward and reverse sequence pairs were merged via overlapping regions and the VHHs, including CDRs, were annotated. Based on CDR3 identity, V-body sequences were clustered, allowing for a detailed analysis of V-body enrichment during phage display, sequence diversity, CDRS length distribution and cluster abundance. Based on these analyses, up to 500 candidates were selected for DNA synthesis by Twist and further characterization.

[0164] Figures 4A-4B illustrate on-cell binding of anti-TRAILR2 V-body monomers to an engineered TRAILR2 overexpressing cell line.

[0165] Figures 5A-5B illustrate on-cell binding of anti-TRAILR2 V-body monomers to an endogenous TRAILR2 expressing cell line.

[0166] Figure 6 shows a schematic diagram of an exemplary experimental setup for determination of binding affinities of the V-bodies for their respective target via surface plasmon resonance (SPR). Figure discloses " HHHHHH" as SEQ ID NO: 2407.

[0167] Figures 7A-7G depict surface plasmon resonance (SPR) sensorgrams of VHH binding to human, cynomolgus (cyno), and mouse TRAILR2. Fitted binding curves and calculated dissociation constants (KD) are included. The first panel shows binding affinities of single V-bodies to human TRAILR2. Each tile represents the global analysis of human TRAILR2 (extracellular domain) binding to a single V-body captured onto a discrete spot, with KDvalues reported. The lines representing ascending TRAILR2 concentration are fitted globally indicating that the binding kinetics are well-described by a simple Langmuir mode. Spots with non-ideal behaviors are boxed (e.g., insufficient on-rate / off-rate information, inactive or barely binding). Binding affinities were measured under physiological conditions (50 mM HEPES pH 7.4, 150 mM NaCI, 0.1 % (w / v) BSA, 0.05% (v / v) Tween20, 25°C) using 11 different antigen concentrations (2-fold serial dilution, starting at 1 pM). The second panel shows binding affinities of single V-bodies to cyno TRAILR2 extracellular domain. The third panel shows binding affinities of single V-bodies to mouse TRAILR2 extracellular domain. Two competitor molecules were used as controls in Figure 7G. Figure discloses " HHHHHH" as SEQ ID NO: 2407.

[0168] Figure 8 shows a schematic diagram of an exemplary experimental setup for determination of ligand competition with TRAIL.

[0169] Figures 9A-9I depict surface plasmon resonance (SPR) sensorgrams for determination of TRAIL blocking. Each tile represents the sensorgram overlay plot for a single V-body captured onto a discrete spot. In Figures 9A-9G, the solid-lined sensorgram displays TRAIL competition: association of human TRAILR2 (extracellular domain) followed either by additional binding by TRAIL, indicating an unoccupied41323195900v2Attorney Docket No: 260525.000128epitope (non-overlapping epitopes), or no TRAIL binding, indicating epitope blocking (overlapping epitopes); while the dotted-lined sensogram displays the buffer control: association and dissociation of human TRAILR2 in the absence of TRAIL). In Figures 9H-9I, the ligand and buffer control sensograms are as indicated. Human TRAILR2 (extracellular domain) was injected (500 nM) under physiological conditions (50 mM HEPES pH 7.4, 150 mM NaCI, 0.1 % (w / v) BSA, 0.05% (v / v) Tween20, 25°C) followed by human TRAIL (1 |1M).

[0170] Figure 10 depicts potency of necrosis induction by TRAILR2 V-body agonists of distinct valency in an endogenous TRAILR2 expressing cell line, compared to a tetravalent competitor molecule (competitor #1 in SPR characterization data).

[0171] Figure 11 depicts the time course of Annexin V induction by TRAILR2 V-body agonists of distinct valency in an endogenous TRAILR2 expressing cell line.

[0172] Figures 12A-12B depict that distinct tetravalent formats display comparable kinetics and potency.

[0173] Figure 13 illustrates exemplary designs of multivalent anti-TRAILR2 V-body constructs.Different V-body formats for testing of TRAILR2 signal induction are depicted. Anti-TRAILR2 V-bodies are shown as oval shapes, linkers are shown in lines with Glycine-Serine (GS) linkers as curved lines, rigid linkers as straight lines and Fc domains as dimeric bars. A first set of constructs comprises anti-TRAILR2 V-bodies connected via flexible GS linkers. A second set of constructs with the same anti-TRAILR2 valencies comprises a mixture of flexible GS linkers and a rigid linker. A rigid proline-rich linker included in some of the constructs can mimic the distance of "90-100A that can be spanned by a conventional antibody (e.g., a monoclonal antibody, mAb) and roughly equals the size of the death-inducing signaling complex (DISC) complex. V-bodies fused to a silenced antibody Fc domain are also included. The rigidity of an Fc domain and the distance spanned indicates its suitability for TRAILR2 signal induction. A set of Fc mounted V-bodies have also been designed, including N- and C-terminal fusions of V-bodies to the Fc domain with different valencies. Figure discloses " PAPAP" as SEQ. ID NO: 2408. Similarly multivalent anti-CDH17 V-body constructs can also be designed.

[0174] Figure 14 depicts high throughput kinetic analysis of T00022Hul and T00007Hul. Binding affinities of human TRAILR2 are shown in the top panel. Each tile represents the global analysis of human TRAILR2 (extracellular domain) binding to a single V-body captured onto a discrete spot, with KD values reported. The lines representing ascending TRAILR2 concentration were fitted globally, indicating that the binding kinetics were well-described by a simple Langmuir mode. Binding affinities were measured under physiological conditions (50 mM HEPES pH 7.4, 150 mM NaCI, 0.1 % (w / v) BSA, 0.05%42323195900v2Attorney Docket No: 260525.000128(v / v) Tween20, 25°C) using 10 different antigen concentrations (2-fold serial dilution, starting at 200 nM (human TRAILR2), 300 nM (cyno TRAILR2) or 1 |1M (mouse TRAILR2)). Binding affinities of cyno TRAILR2 are shown in the middle panel. Binding affinities of mouse TRAILR2 are shown in the bottom panel. Figure discloses " HHHHHH" as SEQ ID NO: 2407.

[0175] Figure 15 depicts high throughput kinetic analysis of T00023Hul and T00024Hul. Binding affinities of human TRAILR2 are shown in the top panel. Each tile represents the global analysis of human TRAILR2 (extracellular domain) binding to a single V-body captured onto a discrete spot, with KDvalues reported. The lines representing ascending TRAILR2 concentration are fitted globally, indicating that the binding kinetics were well-described by a simple Langmuir mode. Binding affinities were measured under physiological conditions (50 mM HEPES pH 7.4, 150 mM NaCI, 0.1 % (w / v) BSA, 0.05% (v / v) Tween20, 25°C) using 10 different antigen concentrations (2-fold serial dilution, starting at 62.5 nM (human TRAILR2), 125 nM (cyno TRAILR2) or 1 |1M (mouse TRAILR2)). Binding affinities of cyno TRAILR2 are shown in the middle panel. Binding affinities of mouse TRAILR2 are shown in the bottom panel. Figure discloses " HHHHHH" as SEQ ID NO: 2407.

[0176] Figure 16 shows off-target binding analysis of T00013Hul, T00024Hul, T00007Hul, T00022Hul, and T00023Hul. Binding affinities of V-bodies for human TRAILR2 compared to homolog proteins TRAILR1, TRAILR3 and TRAILR4 were assessed. Each tile represents the global analysis of a receptor binding to a single V-body captured onto a discrete spot, with KDvalues reported. The lines representing ascending receptor concentrations were fitted globally, indicating that the binding kinetics were well-described by a simple Langmuir mode. Spots with non-ideal behaviors are displayed in boxes (e.g., insufficient on-rate / off-rate information, inactive or barely binding). Binding affinities were measured under physiological conditions (50 mM HEPES pH 7.4, 150 mM NaCI, 0.1 % (w / v) BSA, 0.05% (v / v) Tween20, 25°C) using 12 different antigen concentrations (2-fold serial dilution, starting at 500 nM for TRAILR1, TRAILR3 and TRAILR4, and starting at 31.3 nM for TRAILR2).

[0177] Figure 17 shows constructs for V-body domain mapping. Swap constructs were generated to identify the TRAILR2 domain that was targeted by different V-bodies. Individual cysteine-rich domains (CRDs) or the stalk region were swapped from the human protein sequence to the camelid protein sequence to generate hybrid TRAILR2 extracellular receptor domains (ECDs).

[0178] Figures 18A-18B illustrate domain mapping of TRAILR2 binding V-bodies T00007Hul, T00015HU1, T00022HU1, T00003Hul, T00008Hul, and T00012Hul (Figure 18A), and T00023Hul and T00024Hul (Figure 18B). Binding of V-bodies to TRAILR2 human / camelid swap extracellular receptor domain (ECD) proteins was determined. Each tile represents the global analysis of TRAILR2 binding to a43323195900v2Attorney Docket No: 260525.000128single V-body captured onto a discrete spot. The lines represent ascending receptor concentrations injected. Spots where no binding event was observed, when compared to the WT TRAILR2 ECD sensorgram, are displayed in boxes.

[0179] Figures 19A-19B show epitope binning of TRAILR2 binding V-bodies. Simultaneous binding of two V-bodies to TRAILR2 was determined to test if the V-bodies had overlapping epitopes. V-bodies were coupled to the SPR chip, saturated with TRAILR2 and tested for binding of an additional V-body that was injected. An example of an experimental set for competition binding and epitope binning analysis is depicted in Figure 19A. Results of the binning analysis are shown in a matrix representation (Figure 19B). Dotted squares indicate that no simultaneous binding of the 2 V-bodies was possible, showing that their epitope on TRAILR2 is overlapping. Striped squares indicate that a simultaneous binding of the tested V-bodies was possible. Hence, these V-bodies target distinct epitopes on TRAILR2.

[0180] Figures 20A-20G show kinetic analysis of CDR3 alanine (Ala) scan V-body mutants. Binding affinities of CDR3 alanine (Ala) scan mutant V-bodies for human TRAILR2 were determined for Ala scan mutants T00012Hul (Figures 20A-20B), T00007Hul (Figures 20C-20D), T00023Hul (Figure 20E), T00024Hul (Figure 20F), and T00022Hul (Figure 20G). Each tile represents the global analysis of human or cyno TRAILR2 binding to a single V-body captured onto a discrete spot, with KD values reported. The lines representing ascending receptor concentrations were fitted globally, indicating that the binding kinetics were well-described by a simple Langmuir mode. Spots with non-ideal behaviors are displayed in boxes (e.g., insufficient on-rate / off-rate information, inactive or barely binding). Binding affinities were measured under physiological conditions (50 mM HEPES pH 7.4, 150 mM NaCI, 0.1 % (w / v) BSA, 0.05% (v / v) Tween20, 25°C) using 7 different antigen concentrations (2-fold serial dilution, starting at 15.6 nM for human and cyno TAILR2).

[0181] Figures 21A-21E display on-cell binding fluorescence-activated cell sorting (FACS) analysis of CDR3 alanine (Ala) scan V-body mutants T00012Hul (Figure 21A), T00007Hul (Figure 21B), T00023Hul (Figure 21C), T00024Hul (Figure 21D), and T00022Hul (Figure 21E). All data are represented as percent mean fluorescence intensity (MFI) compared to parental V-body. Figure discloses SEQ ID NOS 2328, 2327, 2326, 2325, 2324, 2323, 2322, 2321, 2320, 2319, 2318, 2317, 2316, 2315, 2314, 2313, 2312, 2311, 2310, 2309, 2308, 2307, 2306, 2305, 2304, 2303, 2302, 2301, 2300, 2299, 2298, 2344, 2343, 2342, 2341, 2340, 2339, 2338, 2348, 2347, 2346, 2345, 2337, 2336, 2335, 2334, 2333, 2332, 2331, 2330 and 2329, respectively, in order of appearance.

[0182] Figure 22 depicts an exemplary immune library construction and general panning strategy for discovery of CDH17 binders.44323195900v2Attorney Docket No: 260525.000128

[0183] Figure 23 shows sample selection for next generation sequencing (NGS) throughout the phage display process. For the three initial libraries, 6 CDH17 samples of the first panning round, and 12 CDH17 samples of the second panning round were sequenced with 20 million, 2 million, and 2 million reads, respectively. Comparison of V-body enrichment from the initial library to the first and second round of panning enabled the identification of potential V-body candidates.

[0184] Figure 24 shows a schematic diagram of an exemplary NGS workflow. Following phage display, the VHH region of the phage elutions was polymerase chain reaction (PCR) amplified, unique and sample-specific barcodes were fused, and NGS was performed with the Illumina NovaSeq platform from Genewiz. The raw data were demultiplexed, and then processed by the Pipebio NGS analysis pipeline. Forward and reverse sequence pairs were merged via overlapping regions and the VHHs, including CDRs, were annotated. Based on CDRS identity, V-body sequences were clustered, allowing for a detailed analysis of V-body enrichment during phage display, sequence diversity, CDR3 length distribution and cluster abundance. Based on these analyses, up to about 500 candidates were selected for DNA synthesis by Twist and further characterization.

[0185] Figure 25 shows a schematic domain representation and structure prediction of CDH17.Amino acid positions are indicated for the extracellular, transmembrane, and intracellular domains. Extracellular cadherin (EC) repeats are shown positioned superior to the membrane. N-glycans and calcium-binding sites are shown as stars and black dots, respectively. The structure prediction of CDH17 was downloaded from alphafold.ebi.ac.uk / . Information about the amino acid position and domain organization were extracted from Uniport.

[0186] Figures 26A-26E illustrate on-cell binding data. Figure 26A illustrates on-cell binding of anti-CDH17 V-body monomers to GP2d cell line. Figure 26B illustrates on-cell binding of anti-CDH17 V-body monomers to Colo205 cell line. Figure 26C illustrates on-cell binding of anti-CDH17 V-body monomers to DLD-1 cell line. Figure 26D illustrates on-cell binding of anti-CDH17 V-body monomers to T84 cell line.Figure 26E illustrates on-cell binding of anti-CDH17 V-body monomers to HCT-15 cell line.

[0187] Figure 27 shows a schematic diagram of an exemplary experimental setup for determination of epitope binning. Figure discloses " HHHHHH" as SEQ ID NO: 2407.

[0188] Figure 28 shows epitope binning of the V-bodies.

[0189] Figure 29 shows a summary of fluorescence activated cell sorting (FACS) domain mapping.

[0190] Figures 30A-30F illustrate non-limiting engineering principles of anti-TRAILR2 agonist development and exemplary multispecific antibody-binding proteins encompassed by the present invention. Figure 30A illustrates engineering of cancer cell-selective TRAILR2 agonists. Figures 30B-30F45323195900v2Attorney Docket No: 260525.000128show an overview of exemplary engineering formats of multispecific antigen-binding proteins. Figure 30F is adapted from C. Spiess et al. Molecular Immunology 67 (2015) 95-106, which is incorporated herein by reference in its entirety.

[0191] Figures 31A-31X illustrate on-cell binding data. Figures 31A-31H illustrate on-cell binding of bispecific anti-TRAILR2 and anti-CDH17 constructs to the GP2d cell line. Figures 31I-31P illustrate on-cell binding of bispecific anti-TRAILR2 and anti-CDH17 constructs to the HEK293 TRAILR2 overexpressing cell line. Figure 31Q-31X illustrate the lack of binding of bispecific anti-TRAILR2 and anti-CDH17 constructs to target-negative CHO-K1 Cells.

[0192] Figures 32A-32G illustrate cell titer glow (CTG) functional assay data. Figures 32A-32D illustrate the cell viability of the GP2d cells after incubation with bispecific anti-TRAILR2 and anti-CDH17 constructs for 48 hours. Figures 32E-32G illustrate the cell viability of DLD-1 cells after incubation with bispecific anti-TRAILR2 and anti-CDH17 constructs for 48 hours.

[0193] Figures 33A-33D illustrate CDH17-dependent agonist activity in CDH17 expressing GP2d (Figures 33A and 33C) and CDH17-knock out GP2d (Figures 33B and 33D).

[0194] Figure 34 illustrates the cell viability of human hepatocytes after incubation with bispecific anti-TRAILR2 and anti-CDH17 constructs in a hepatotoxicity assessment.

[0195] Figure 35 illustrates the results of a single dose pharmacokinetics study.

[0196] Figures 36A-36D illustrate the in vivo efficacy of bispecific anti-TRAILR2 and anti-CDH17 antigen-binding proteins. Figure 36A shows the efficacy of a single dose of exemplary bispecific constructs in the GP2d cell line. Figure 36B shows the dose response efficacy of exemplary bispecific constructs in the GP2d cell line. Figure 36C shows the dose response efficacy of exemplary bispecific constructs in the DLD-1 cell line. Figure 36D shows the repeat dosing efficacy of exemplary bispecific constructs in the DLD-1 and Colo205 cell lines.

[0197] Figure 37 depicts that TRAILR2 is a death receptor that activates the extrinsic apoptotic pathway, resulting in apoptotic cell death. This figure was adapted from Nature Reviews Cancer 2017;17(6):352, which is incorporated herein by reference in its entirety.

[0198] Figure 38 shows that TC0124 induces superior maximal cell kill in colorectal cancer (CRC) cell lines. Cell lines were treated with TRAILR2 agonists starting at 250 nM with 5-fold dilution for 12 points; cell viability was assessed by CTG at 24, 48, and 72 hours, normalized to non-treated cells, and represented as relative cell death; representative data at 72 hours are shown (n = 5).

[0199] Figure 39 shows that TC0124 displays CDH17-dependent agonist activity and cell kill. CRC cell lines were treated with TRAILR2 agonists starting at 250 nM with 5-fold dilution for 12 points; cell46323195900v2Attorney Docket No: 260525.000128viability was assessed by CTG at 24 h, 48 h, and 72 h, normalized to non-treated cells, and represented as relative cell death; representative data at 72 h shown (n = 5).

[0200] Figure 40 shows that TC0124 has no activity against primary human hepatocytes. Cells were treated for 48 hours with agonist in presence of 1 mg / mL I Vlg; full dose response starting at 100 nM with 10-fold dilution across 12 points; representative data shown for 10 nM-10 fM; n = 2 for Comparator 4 and n = 5 for all other molecules.

[0201] Figures 41A-41C show that TC0124 has efficacy in xenograft models of CRC cell lines with high and intermediate sensitivity to TRAILR2 agonists. TC0124 has comparable single-dose efficacy to Comparator 1 in the highly sensitive GP2d xenograft (Figure 41A). TC0124 achieves higher rate of partial responses in the intermediate sensitive DLD-1 model compared to Comparator 1 (Figure 41B) and Comparator 4 (Figure 41C). PR# = partial response defined as two consecutive measurements where tumor volume is below initial measurement from the start of dosing; * statistical significance was determined by an ordinary one-way ANOVA followed by Tukey's multiple comparisons test (p < 0.05).

[0202] Figure 42 shows enhanced activity of TC0124 in combination with chemotherapy in CRC cell lines.

[0203] Figure 43 shows biophysical characterization of TC0124.DETAILED DESCRIPTION

[0204] Apoptosis induction in cancer cells has long been considered an attractive strategy to treat solid tumors. TRAILR2, also known as death receptor 5, is expressed on the cell surface and receptor clustering triggers downstream assembly of a death inducing signaling complex (DISC) and activation of caspases leading to cell death (Nature Reviews Cancer 2017;17(6):352; Mol Cancer Ther 2022;21:594)

[0205] TRAILR2 expression is upregulated in cancer cells and has been explored clinically with limited success. Suboptimal clustering of TRAILR2 receptors, leading to insufficient apoptotic signaling limited the efficacy of first generation TRAILR2 agonists while second-generation, multivalent, non-targeted TRAILR2 agonists, have been limited by hepatotoxicity due to high liver expression of TRAILR2 (Cancer Chemother Pharmacol (2015) 75:887-895b).Definitions

[0206] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. For47323195900v2Attorney Docket No: 260525.000128purposes of interpreting this specification, the following description of terms will apply and whenever appropriate, terms used in the singular will also include the plural and vice versa. All patents, applications, published applications and other publications are incorporated by reference in their entirety. In the event that any description of terms set forth conflicts with any document incorporated herein by reference, the description of term set forth below shall control.

[0207] As used herein, the term "about" when used in reference to a particular recited numerical value, means that the value may vary from the recited value by no more than 5%. For example, as used herein, the expression "about 100" includes 95 and 105 and all values in between (e.g., 96, 97, 98, 99, etc.).

[0208] The term "antigen" encompasses any agent (e.g., protein, peptide, polysaccharide, glycoprotein, glycolipid, nucleotide, portions thereof, or combinations thereof) that may be specifically bound by the products of specific humoral or cellular immunity, such as an antibody molecule or T-cell receptor. In various embodiments of the present disclosure, the antigen described herein is TRAILR2, including human, cynomolgus, and / or mouse TRAILR2, and / or CDH17, including human, cynomolgus, and / or mouse CDH17.

[0209] The term "epitope" can refer to an antigenic determinant on the surface of an antigen to which an antibody molecule binds. A single antigen may have more than one epitope. Thus, different antibodies may bind to different areas on an antigen and may have different biological effects (e.g., agnostic or antagonistic effects). Epitopes may be either conformational or linear. A conformational epitope is formed by spatially juxtaposed amino acids from different segments of the linear polypeptide chain. A linear epitope is formed by adjacent amino acid residues in a polypeptide chain. In some cases, an epitope may include non-peptidic moieties on the antigen, such as saccharides, phosphoryl groups, or sulfonyl groups.

[0210] The term "antigen-binding protein" refers in its broadest sense to a protein that specifically binds an antigen (e.g., TRAILR2 and / or CDH17). In certain embodiments, an antigen-binding protein is an antibody or an antigen-binding fragment of an antibody, such as a human antibody, a humanized antibody; a camelid antibody; a chimeric antibody; a recombinant antibody; a heavy chain antibody; a single-domain antibody (e.g., VHH); a single chain antibody (e.g., single chain fragment variable (scFv)); a diabody; a triabody; a tetrabody; a Fab fragment; a F(ab') 2 fragment; an IgD antibody; an IgE antibody; an IgM antibody; an IgGl antibody; an lgG2 antibody; an lgG3 antibody; or an lgG4 antibody, and fragments thereof. The term "antigen-binding protein" also encompasses, for example, an alternative protein scaffold or artificial scaffold with grafted CDRs or CDR derivatives. Such scaffolds include, but are48323195900v2Attorney Docket No: 260525.000128not limited to, antibody-derived scaffolds comprising mutations introduced to, for example, stabilize the three-dimensional structure of the antigen-binding protein as well as wholly synthetic scaffolds comprising, for example, a biocompatible polymer. In addition, peptide antibody mimetics can be used, as well as scaffolds based on antibody mimetics utilizing fibronectin components (e.g., fibronectin type III domain ( FN3 )) as a scaffold.

[0211] The term "antigen-binding domain" refers to the portion of the antigen-binding protein that is capable of specifically binding to an antigen or epitope. An antigen-binding protein may comprise more than one antigen-binding domains, for example, two, three, four, five, six, seven, eight or more antigenbinding domains. For example, antigen-binding domains may comprise at least one variable region (either a heavy chain or light chain variable region) or one or more CDRs of an antibody that can bind a particular antigen. Examples of suitable antigen-binding domains include, without limitation, singledomain antibodies {e.g., VHH), single-chain antibodies (e.g., single chain fragment variable (scFv)), Fab fragments, F(ab')2 fragments, and protein scaffolds.

[0212] The term " TNF-related apoptosis-inducing ligand receptor 2", or " TRAILR2", or the like, are used interchangeably herein and can refer to any isoform(s), variant(s), and / or species homolog(s) of TRAILR2 from any source, e.g., mammals including primates (e.g., humans and monkeys) and rodents (e.g., rats and mice). The term encompasses naturally-occurring variants of TRAILR2 such as but not limited to allelic variants and splice variants. The term also encompasses "full-length" or unprocessed TRAILR2 in addition to any form of TRAILR2 that can result from processing such as that which may occur within a cell. In some embodiments, TRAILR2 is human TRAILR2. In some embodiments, TRAILR2 is cynomolgus monkey ("cyno") TRAILR2. In some embodiments, TRAILR2 is mouse TRAILR2.

[0213] The term "cadherin 17", or " CDH17", or the like, are used interchangeably herein and can refer to any isoform(s), variant(s), and / or species homolog(s) of CDH17 from any source, e.g., mammals including primates (e.g., humans and monkeys) and rodents (e.g., rats and mice). The term encompasses naturally-occurring variants of CDH17 such as but not limited to allelic variants and splice variants. The term also encompasses "full-length" or unprocessed CDH17 in addition to any form of CDH17 that can result from processing such as that which may occur within a cell. In some embodiments, CDH17 is human CDH17. In some embodiments, CDH17 is cynomolgus monkey ("cyno") CDH17. In some embodiments, CDH17 is mouse CDH17.

[0214] The term "antibody" and "immunoglobulin" or " Ig" are used interchangeably herein, and is used in the broadest sense and encompasses, for example, individual monoclonal antibodies (including agonist, antagonist, neutralizing antibodies, full length or intact monoclonal antibodies), antibody49323195900v2Attorney Docket No: 260525.000128compositions with polyepitopic or monoepitopic specificity, polyclonal antibodies, monovalent antibodies, multivalent antibodies, multispecific antibodies (e.g., bispecific antibodies), single-domain antibodies (e.g., VHH), single chain antibodies, intrabodies, anti-idiotypic (anti-ld) antibodies, and antigen-binding fragments of antibodies, as described below. An antibody can be human, humanized, camelized, recombinantly produced, chimeric, synthetic, affinity de-matured and / or affinity matured as well as an antibody from other species, for example mouse, camel, llama, rabbit, etc. In specific embodiments, the specific target antigen that can be bound by an antibody provided herein includes a TRAILR2 polypeptide, TRAILR2 fragment or TRAILR2 epitope. In specific embodiments, the specific target antigen that can be bound by an antibody provided herein includes a CDH17 polypeptide, CDH17 fragment or CDH17 epitope. An "antigen-binding fragment" generally refers a portion of an antibody heavy and / or light chain polypeptide that retains some or all of the binding activity of the antibody from which the fragment was derived. Non-limiting examples of antigen-binding fragments include singledomain antibody (e.g., VHH), single-chain Fvs (scFv), Fab fragments, F(ab') fragments, F(ab)2 fragments, F(ab')2 fragments, disu If ide-l i n ked Fvs (sdFv), Fd fragments, Fv fragments, diabody, triabody, tetrabody and minibody, or a chemically modified derivative thereof. In particular, antibodies provided herein include immunoglobulin molecules and molecules that contain immunologically active portion(s) of an immunoglobulin molecule, for example, one or more complementarity determining regions (CDRs) of an antibody that binds to TRAILR2 and / or CDH17. Such antibody fragments can be found described in, for example, Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, New York (1989); Myers (ed.), Molec. Biology and Biotechnology: A Comprehensive Desk Reference, New York: VCH Publisher, Inc.; Huston et al., Cell Biophysics, 22:189-224 (1993); Pliickthun and Skerra, Meth. Enzymol., 178:497-515 (1989) and in Day, E. D., Advanced Immunochemistry, Second Ed., Wiley-Liss, Inc., New York, N. Y. (1990). The antibodies provided herein can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), any class (e.g., IgGl, lgG2, lgG3, lgG4, IgAl and lgA2), or any subclass (e.g., lgG2a and lgG2b) of immunoglobulin molecule.

[0215] The term "single-domain antibody" or "sdAb" as used herein, refers to an antibody or antibody fragment containing a single antibody variable domain that is able to bind to a specific antigen alone, without the requirement of another antibody variable domain. The complementarity determining regions (CDRs) of a single-domain antibody are part of a single antibody variable domain. Examples of single-domain antibodies include, but are not limited to, heavy chain antibodies, antibodies naturally devoid of light chains, single domain antibodies derived from conventional four-chain antibodies, engineered antibodies, variable domains derived from the aforementioned antibodies, and single50323195900v2Attorney Docket No: 260525.000128domain scaffolds other than those derived from antibodies. Single domain antibodies may be derived from any species including, but not limited to mouse, human, camel, llama, shark, goat, rabbit, and / or bovine. In some embodiments, a single domain antibody as used herein is a naturally occurring single domain antibody known as heavy chain antibody devoid of light chains. For clarity reasons, the variable domain derived from a heavy chain antibody naturally devoid of light chain is known herein as a VHH to distinguish it from the conventional VH of four-chain immunoglobulins. Such a VHH molecule can be derived from antibodies raised in Camelidae species, e.g., camel, llama, dromedary, alpaca and guanaco. Other species besides Camelidae may produce heavy chain antibodies naturally devoid of light chain, which are also within the scope of the invention. For example, cartilaginous fishes such as sharks can produce immunoglobulin-like structures known as VNAR. In some embodiments, a single-domain antibody may be obtained from a Camelidae VH domain. In some embodiments, a single-domain antibody may be obtained from human VH by camelization. See Saerens et al., Current Opinion in Pharmacology, 2008, 8:600-608, the disclosure of which being incorporated by reference, for review of single-domain antibodies.

[0216] The term "specifically binds" as used herein means that an antigen-binding protein forms a complex with a target antigen that is relatively stable under physiologic conditions. Specific binding can be characterized by a dissociation constant (KD) of about 1x10sM or less (e.g., less than 10sM, less than 5xl0-7M, less than 10'7M, less than 5xl0‘8M, less than 10'8M, less than 5xl0-9M, less than 10'9M, or less than 1010M). Methods for determining the binding affinity of an antigen-binding protein, e.g., an antibody or an antibody fragment, to a target antigen are well known in the art and include, e.g., surface plasmon resonance (e.g., BIACORE® assays), bio-layer interferometry, ligand binding assays (e.g., enzyme-linked immunosorbent assay (ELISA)), equilibrium dialysis, fluorescent-activated cell sorting (FACS), or flow cytometry-based binding assays and the like. Specific binding to a particular target antigen from a certain species does not exclude that the antigen-binding protein can also specifically bind to the analogous target from a different species. For example, specific binding to human TRAILR2 does not exclude that the antigen-binding protein can also specifically bind to TRAILR2 from cynomolgus monkeys ("cyno") or mice.

[0217] The term "isolated" when used in the context of antigen-binding proteins (e.g., antibodies, such as single-domain antibodies), polypeptides, polynucleotides, and vectors, means the antigenbinding proteins (e.g., antibodies, such as single-domain antibodies), polypeptides, polynucleotides and vectors are at least partially free of other biological molecules from the cells or cell culture from which they are produced. Such biological molecules include nucleic acids, proteins, other antibodies or51323195900v2Attorney Docket No: 260525.000128antigen-binding fragments, lipids, carbohydrates, or other material such as cellular debris and growth medium. An isolated antigen-binding protein may further be at least partially free of expression system components such as biological molecules from a host cell or of the growth medium thereof. Generally, the term "isolated" is not intended to refer to a complete absence of such biological molecules (e.g., minor or insignificant amounts of impurity may remain) or to an absence of water, buffers, or salts or to components of a pharmaceutical formulation that includes the antigen-binding proteins (e.g., antibodies, such as single-domain antibodies).

[0218] The term "operably linked" as used herein can refer to a functional relationship between two or more regions of a polypeptide chain in which the two or more regions are linked so as to produce a functional polypeptide.

[0219] As used herein, the term "variant", "derivative" or "derived from" in the context of proteins or polypeptides (e.g., antigen-binding proteins or domains thereof) refer to: (a) a polypeptide that has at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% sequence identity to the polypeptide it is a variant or derivative of; (b) a polypeptide encoded by a nucleotide sequence that has at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98% or 99% sequence identity to a nucleotide sequence encoding the polypeptide it is a variant or derivative of; (c) a polypeptide that contains 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acid mutations (i.e., additions, deletions and / or substitutions) relative to the polypeptide it is a variant or derivative of; (d) a polypeptide encoded by nucleic acids can hybridize under high, moderate or typical stringency hybridization conditions to nucleic acids encoding the polypeptide it is a variant or derivative of; (e) a polypeptide encoded by a nucleotide sequence that can hybridize under high, moderate or typical stringency hybridization conditions to a nucleotide sequence encoding a fragment of the polypeptide, it is a variant or derivative of, of at least 20 contiguous amino acids, at least 30 contiguous amino acids, at least 40 contiguous amino acids, at least 50 contiguous amino acids, at least 75 contiguous amino acids, at least 100 contiguous amino acids, at least 125 contiguous amino acids, or at least 150 contiguous amino acids; or (f) a fragment of the polypeptide it is a variant or derivative of. The terms also encompass a fusion protein or polypeptide comprising the polypeptide it is a variant or derivative of.

[0220] The term "substantial identity" or "substantially identical", when referring to a nucleic acid or fragment thereof, indicates that, when optimally aligned with appropriate nucleotide insertions or deletions with another nucleic acid (or its complementary strand), there is nucleotide sequence identity in at least about 95%, and more preferably at least about 96%, 97%, 98% or 99% of the nucleotide bases,52323195900v2Attorney Docket No: 260525.000128as measured by any well-known algorithm of sequence identity, such as FAST A, BLAST or Gap, as discussed below. A nucleic acid molecule having substantial identity to a reference nucleic acid molecule may, in certain instances, encode a polypeptide having the same or substantially similar amino acid sequence as the polypeptide encoded by the reference nucleic acid molecule.

[0221] As applied to polypeptides, the term "substantial similarity" or "substantially similar" means that two peptide sequences, when optimally aligned, such as by the programs GAP or BESTFIT using default gap weights, share at least 95% sequence identity, even more preferably at least 98% or 99% sequence identity. Preferably, residue positions which are not identical differ by conservative amino acid substitutions. A "conservative amino acid substitution" is one in which an amino acid residue is substituted by another amino acid residue having a side chain (R group) with similar chemical properties (e.g., charge or hydrophobicity). In general, a conservative amino acid substitution will not substantially change the functional properties of a protein. In cases where two or more amino acid sequences differ from each other by conservative substitutions, the percent sequence identity or degree of similarity may be adjusted upwards to correct for the conservative nature of the substitution. Means for making this adjustment are well-known to those of skill in the art. See, e.g., Pearson (1994) Methods Mol. Biol. 24: 307-331, herein incorporated by reference. Examples of groups of amino acids that have side chains with similar chemical properties include (1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; (2) aliphatic-hydroxyl side chains: serine and threonine; (3) amide-containing side chains: asparagine and glutamine; (4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; (5) basic side chains: lysine, arginine, and histidine; (6) acidic side chains: aspartate and glutamate, and (7) sulfur-containing side chains are cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamateaspartate, and asparagine-glutamine. Alternatively, a conservative replacement is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256: 1443-1445, herein incorporated by reference. A "moderately conservative" replacement is any change having a nonnegative value in the PAM250 log-likelihood matrix.

[0222] Sequence similarity for polypeptides, which is also referred to as sequence identity, is typically measured using sequence analysis software. Protein analysis software matches similar sequences using measures of similarity assigned to various substitutions, deletions and other modifications, including conservative amino acid substitutions. For instance, GCG software contains programs such as Gap and Bestfit which can be used with default parameters to determine sequence homology or sequence identity between closely related polypeptides, such as homologous polypeptides from different species53323195900v2Attorney Docket No: 260525.000128of organisms or between a wild-type protein and a mutein thereof. See, e.g., GCG Version 6.1.Polypeptide sequences also can be compared using FASTA using default or recommended parameters, a program in GCG Version 6.1. FASTA (e.g., FASTA2 and FASTA3) provides alignments and percent sequence identity of the regions of the best overlap between the query and search sequences (Pearson (2000) supra). Another preferred algorithm when comparing a sequence of the disclosure to a database containing a large number of sequences from different organisms is the computer program BLAST, especially BLASTP or TBLASTN, using default parameters. See, e.g., Altschul et al. (1990) J. Mol. Biol. 215:403-410 and Altschul et al. (1997) Nucleic Acids Res. 25:3389-402, each herein incorporated by reference.

[0223] The terms "enhance" or "promote," or "increase," or "expand," or "improve" refer generally to the ability of a composition contemplated herein to produce, elicit, or cause a greater physiological response (i.e., downstream effects) compared to the response caused by either vehicle or a control molecule / composition. A measurable physiological response may include an increase in immune cell expansion, activation, effector function, persistence, and / or an increase in tumor cell death killing ability, among others apparent from the understanding in the art and the description herein. In certain embodiments, an "increased" or "enhanced" amount can be a "statistically significant" amount, and may include an increase that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 or more times (e.g., 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) the response produced by vehicle or a control composition.

[0224] The terms "decrease" or "lower," or "lessen," or "reduce," or "abate" refer generally to the ability of composition contemplated herein to produce, elicit, or cause a lesser physiological response (i.e., downstream effects) compared to the response caused by either vehicle or a control molecule / composition. In certain embodiments, a "decrease" or "reduced" amount can be a "statistically significant" amount, and may include a decrease that is 1.1, 1.2, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30 or more times (e.g., 500, 1000 times) (including all integers and decimal points in between and above 1, e.g., 1.5, 1.6, 1.7. 1.8, etc.) the response (reference response) produced by vehicle or a control composition.

[0225] The terms "treat" or "treatment" of a state, disease, disorder or condition include: (1) preventing, delaying, or reducing the incidence and / or likelihood of the appearance of at least one clinical or sub-clinical symptom of the state, disease, disorder or condition developing in a subject that may be afflicted with or predisposed to the state, disease, disorder or condition, but does not yet experience or display clinical or subclinical symptoms of the state, disease, disorder or condition; or (2)54323195900v2Attorney Docket No: 260525.000128inhibiting the state, disease, disorder or condition, / .e., arresting, reducing or delaying the development of the state, disease, disorder or condition or a relapse thereof or at least one clinical or sub-clinical symptom thereof; or (3) relieving the state, disease, disorder or condition, i.e., causing regression of the state, disease, disorder or condition or at least one of its clinical or sub-clinical symptoms. The benefit to a subject to be treated is either statistically significant or at least perceptible to the patient or to the physician.

[0226] The terms "effective amount" or "therapeutically effective amount" refer to a quantity and / or concentration of a composition containing an active ingredient (e.g., a bispecific anti-TRAILR2 and anti-CDH17 antigen-binding protein) that when administered into a patient either alone (i.e., as a monotherapy) or in combination with additional therapeutic agents, yields a significant decrease in the progression of the state, disease, disorder, or condition, as, for example, by ameliorating or eliminating symptoms and / or the cause of the state, disease, disorder, or condition. An effective amount may be an amount that relieves, lessens, or alleviates at least one symptom or biological response or effect associated with a state, disease, disorder, or condition, prevents progression of the state, disease, disorder, or condition, or improves physical functioning of the patient. A therapeutically effective amount of a composition containing an active agent may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the active agent to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the active agent are outweighed by the therapeutically beneficial effects. A therapeutically effective amount may be delivered in one or more administrations. A therapeutically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic and / or prophylactic result.

[0227] The terms "individual", "subject" and "patient" are used interchangeably herein to refer to an animal, for example a mammal. The terms include human and veterinary subjects. In some embodiments, methods of treating mammals, including, but not limited to, humans, rodents, simians, felines, canines, equines, bovines, porcines, ovines, caprines, mammalian laboratory animals, mammalian farm animals, mammalian sport animals, and mammalian pets, are provided. The subject can be male or female and can be any suitable age, including infant, juvenile, adolescent, adult, and geriatric subjects. In some embodiments, a subject can be a subject in need of treatment for a disease or disorder. In particular embodiments, the subject is a human.55323195900v2Attorney Docket No: 260525.000128Multispecific Antigen-Binding Proteins

[0228] In one aspect, provided herein are multispecific antigen-binding proteins comprising one or more anti-TRAILR2 antigen-binding domains and one or more anti CDH17 antigen-binding domains that are linked, directly or indirectly, to one another. The multispecific antigen-binding proteins may comprise one or more additional domains or moieties.

[0229] In some embodiments, the multispecific antigen-binding protein of the present disclosure may be, for example, bispecific, trispecific, tetraspecific, pentaspecific, etc. The terms "bispecific", "trispecific", "tetraspecific", "pentaspecific", etc., all fall under the term "multispecific" and refer to binding to two, three, four, five, etc., different target molecules, respectively. In specific embodiments, the multispecific antigen-binding protein of the present disclosure are bispecific.

[0230] In some embodiments, the multispecific antigen-binding protein of the present disclosure comprises one or more anti-TRAILR2 antigen-binding domains (e.g., single-domain antibodies) and one or more anti CDH17 antigen-binding domains (e.g., single-domain antibodies) as described herein. In some embodiments, the multispecific antigen-binding protein is multivalent. For example, the multispecific antigen-binding protein or conjugate of the present disclosure may be at least bivalent, but can also be e.g., trivalent, tetravalent, pentavalent, hexavalent, septivalent, octavalent, etc. The multispecific antigen-binding protein can be bivalent, trivalent, tetravalent, pentavalent, hexavalent, septivalent, or octavalent for the different antigens. The terms "bivalent", "trivalent", "tetravalent", "pentavalent", "hexavalent", "septivalent", "octavalent" all fall under the term "multivalent" and indicate the presence of two, three, four, five six, seven, or eight binding domains (e.g., single-domain antibodies), respectively.

[0231] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises one or more of an anti-TRAILR2 antigen-binding domain and one or more of an anti-CDH17 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises two or more of an anti-TRAILR2 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises three or more of an anti-TRAILR2 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises four or more of an anti-TRAILR2 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises one, two, three, four, five, six, seven, eight, nine, or ten, or more of an anti-TRAILR2 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises two or more56323195900v2Attorney Docket No: 260525.000128of an anti-CDH17 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises three or more of an anti-CDH17 antigenbinding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises four or more of an anti-CDH17 antigen-binding domain described herein. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises one, two, three, four, five, six, seven, eight, nine, or ten, or more of an anti-CDH17 antigen-binding domain described herein.

[0232] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises at least two, at least three, or at least four anti-TRAILR2 antigen-binding domains, and at least one, at least two, or at least four anti-CDH17 antigen-binding domains.

[0233] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises two anti-TRAILR2 antigen-binding domains, and one anti-CDH17 antigen-binding domain. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises three anti-TRAILR2 antigen-binding domains, and one anti-CDH17 antigen-binding domain. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises four anti-TRAILR2 antigen-binding domains, and one anti-CDH17 antigen-binding domain.

[0234] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises two anti-TRAILR2 antigen-binding domains, and two anti-CDH17 antigen-binding domains. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises three anti-TRAILR2 antigen-binding domains, and two anti-CDH17 antigen-binding domains. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises four anti-TRAILR2 antigen-binding domains, and two anti-CDH17 antigen-binding domains.

[0235] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises two anti-TRAILR2 antigen-binding domains, and four anti-CDH17 antigen-binding domains. In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises four anti-TRAILR2 antigen-binding domains, and four anti-CDH17 antigen-binding domains.

[0236] In some embodiments, the multispecific antigen-binding protein of the present disclosure comprises a single polypeptide. In other embodiments, the multispecific antigen-binding protein or conjugate of the present disclosure comprises more than one polypeptide. In some embodiments, the multispecific antigen-binding protein of the present disclosure comprises two polypeptides.

[0237] In some embodiments, the multispecific antigen-binding protein can comprise an anti-TRAILR2 antigen-binding domain operably connected to an anti-CDH17 antigen-binding domain.57323195900v2Attorney Docket No: 260525.000128

[0238] In various embodiments, the one or more anti-TRAILR2 and / or anti-CDH17 antigen-binding domains can be one or more anti-TRAILR2 and / or anti-CDH17 single-domain antibodies disclosed herein, for example, one or more anti-TRAILR2 and / or anti-CDH17 VHHs disclosed herein.

[0239] When two or more anti-TRAILR2 antigen-binding domains are included in a multispecific antigen-binding protein, the two or more anti-TRAILR2 antigen-binding domains may comprise the same sequence or may comprise different sequences. In such embodiments, the two or more anti-TRAILR2 antigen-binding domains may bind to the same epitope on TRAILR2 or different epitopes on TRAILR2. For example, a multispecific antigen-binding protein or conjugate of the present disclosure may be biparatopic, e.g., if two VHHs bind two different epitopes on TRAILR2.

[0240] In various embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two or more anti-TRAILR2 antigen-binding domains each comprising an amino acid sequence selected from any of the CDR1, CDR2, and / or CDR3 amino acid sequences listed in Table 1-1, or Table 6, or any combination thereof, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity, or any combination thereof. In various related embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two or more anti-TRAILR2 antigen-binding domains each comprising a set of three CDRs ( / .e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-TRAILR2 VHH antibodies listed in Tables 1-1, 1-2, or 6. In some embodiments, the two or more anti-TRAILR2 antigen-binding domains can comprise the same set of three CDRs. In some embodiments, the two or more anti-TRAILR2 antigen-binding domains can comprise different sets of three CDRs.

[0241] When two or more anti-CDH17 antigen-binding domains are included in a multispecific antigen-binding protein or conjugate, the two or more anti-CDH17 antigen-binding domains may comprise the same sequence or may comprise different sequences. In such embodiments, the two or more anti-CDH17 antigen-binding domains may bind to the same epitope on CDH17 or different epitopes on CDH17. For example, a multispecific antigen-binding protein or conjugate of the present disclosure may be biparatopic, e.g., if two VHHs bind two different epitopes on CDH17.

[0242] In various embodiments, a multispecific antigen-binding protein of the present disclosure can comprise two or more anti-CDH17 antigen-binding domains each comprising an amino acid sequence selected from any of the CDR1, CDR2, and / or CDR3 amino acid sequences listed in Table 1-3, or Table 13, or any combination thereof, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity, or any combination thereof. In various related embodiments, a multispecific antigen-binding protein of the present disclosure can comprise two or more anti-CDH1758323195900v2Attorney Docket No: 260525.000128antigen-binding domains each comprising a set of three CDRs ( / .e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-CDH17 VHH antibodies listed in Tables 1-3, 1-4, or 13. In some embodiments, the two or more anti-CDH17 antigen-binding domains can comprise the same set of three CDRs. In some embodiments, the two or more anti-CDH17 antigen-binding domains can comprise different sets of three CDRs.

[0243] In certain embodiments, the one or more additional domain or moieties may be one or more additional binding domain that binds to one or more further antigen or protein, e.g. serum albumin such as human serum albumin (HSA).

[0244] Exemplary designs of bispecific constructs comprising one or more anti-TRAILR2 binding domain {e.g., VHHs) and one or more anti-CDH17 binding domain (e.g., VHHs) are shown in Figures 30B-30F.

[0245] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises a single polypeptide chain, said single polypeptide chain comprising i) one or more antigenbinding domains that specifically bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2); ii) one or more antigen-binding domain(s) that specifically bind to cadherin-17 (CDH17); and iii) optionally, one or more antigen-binding domains that bind human serum albumin (HSA); and optionally, wherein each of the antigen-binding domains is operably linked to each of the other antigen-binding domains via a linker.

[0246] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises a single polypeptide chain comprising a configuration shown in Figures 30B-30C.

[0247] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise one polypeptide chain, said polypeptide chain having the following structure from N-terminus to C-terminus: (anti-TRAILR2 VHH)n- linker - (anti-CDH17 VHH)m, wherein n and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). When n>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. When m>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0248] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise one polypeptide chain, said polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)n- linker - (anti-TRAILR2 VHH)m, wherein n and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). When n>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. When m>2, each anti-TRAILR2 VHH may be59323195900v2Attorney Docket No: 260525.000128optionally operably linked to another anti-TRAILR2 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0249] In some embodiments, the single polypeptide chain comprises three antigen-binding domains (ABDI- ABD3), four antigen-binding domains (ABDI- ABD4), five antigen-binding domains (ABDI- ABD5), or six antigen-binding domains (ABD1-ABD6).

[0250] In some embodiments, the single polypeptide chain comprises three antigen-binding domains, said three antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2; andiii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2.

[0251] In some embodiments, the single polypeptide chain comprises four antigen binding domains, said four antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2; andiv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2.

[0252] In some embodiments, the single polypeptide chain comprises four antigen binding domains, said four antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2; andiv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2.

[0253] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0254] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,60323195900v2Attorney Docket No: 260525.000128ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0255] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds HSA; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0256] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds HSA.

[0257] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds TRAILR2;ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds CDH17;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

[0258] In some embodiments, the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds TRAILR2;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds CDH17; andv) a fifth antigen-binding domain (ABD5) that specifically binds CDH17.61323195900v2Attorney Docket No: 260525.000128

[0259] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0260] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds HSA.

[0261] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds HSA.

[0262] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds HSA; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.62323195900v2Attorney Docket No: 260525.000128

[0263] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds HSA;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0264] In some embodiments, the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N-to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds HSA;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

[0265] In some embodiments, a multispecific antigen-binding protein described herein comprises a first polypeptide chain and a second polypeptide chain, each polypeptide chain comprising:i) one or more antigen-binding domains that specifically bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2),ii) one or more antigen-binding domain that specifically bind to cadherin-17 (CDH17); and iii) an immunoglobulin Fc region; andwherein the first and second polypeptide chains dimerize via the immunoglobulin Fc region to form a dimer, and optionally, wherein each of the antigen-binding domains is operably linked to each of the other antigen-binding domains or the immunoglobulin Fc region via a linker.

[0266] In some embodiments, a multispecific antigen-binding protein of the present disclosure comprises two polypeptide chains, the two polypeptide chains comprising a configuration shown in Figures 30D.

[0267] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-TRAILR2 VHH)n- linker - (anti-CDH17 VHH)m, wherein n and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). When n>2, each anti-TRAILR2 VHH may63323195900v2Attorney Docket No: 260525.000128be optionally operably linked to another anti-TRAILR2 VHH via a linker. When m>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0268] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)n- linker - (anti-TRAILR2 VHH)m, wherein n and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). When n>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. When m>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0269] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-TRAILR2 VHH)n- first linker - Fc region- second linker - (anti-CDH17 VHH)m, wherein n and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). When n>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. When m>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0270] In some embodiments, a multispecific antigen-binding protein or conjugate of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)n- first linker- Fc region - second linker - (anti-TRAILR2 VHH)m, wherein n and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). When n>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. When m>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0271] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)i - (anti-TRAILR2 VHH)n- first linker - Fc region- second linker -(anti-TRAILR2 VHH)m, wherein I, n, and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). The adjacent anti-CDH17 VHH and anti-TRAILR2 VHH may be optionally operably linked to one another via a linker. When I >2, each anti-CDH17 VHH may be optionally operably linked to another anti-64323195900v2Attorney Docket No: 260525.000128CDH17 VHH via a linker. When n>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. When m>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0272] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)i - (anti-TRAILR2 VHH)n- first linker - Fc region- second linker -(anti-CDH17 VHH)m, wherein I, n, and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). The adjacent anti-CDH17 VHH and anti-TRAILR2 VHH may be optionally operably linked to one another via a linker. When I >2, each anti- CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. When n>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. When m>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. The multiple linkers used in the multispecific antigenbinding protein are not necessarily the same or different.

[0273] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-TRAILR2 VHH)i - (anti-CDH17 VHH)n- first linker - Fc region- second linker -(anti-TRAILR2 VHH)m, wherein I, n, and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). The adjacent anti-TRAILR2 VHH and anti-CDH17 VHH may be optionally operably linked to one another via a linker. When I >2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. When n>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. When m>2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. The multiple linkers used in the multispecific antigenbinding protein are not necessarily the same or different.

[0274] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-TRAILR2 VHH)i - (anti-CDH17 VHH)n- first linker - Fc region- second linker -(anti-CDH17 VHH)m, wherein I, n, and m can independently be any integral number (e.g., 1, 2, 3, 4, 5, etc.). The adjacent anti-TRAILR2 VHH and anti-CDH17 VHH may be optionally operably linked to one another via a linker. When I >2, each anti-TRAILR2 VHH may be optionally operably linked to another anti-TRAILR2 VHH via a linker. When n>2, each anti-CDH17 VHH may be optionally operably linked to another anti-CDH17 VHH via a linker. When m>2, each anti-CDH17 VHH may be optionally operably65323195900v2Attorney Docket No: 260525.000128linked to another anti-CDH17 VHH via a linker. The multiple linkers used in the multispecific antigenbinding protein are not necessarily the same or different.

[0275] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: anti-TRAILR2 VHH - first linker - Fc region- second linker - anti-CDH17 VHH. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0276] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: anti-CDH17 VHH - first linker - Fc region- second linker - anti-TRAILR2 VHH. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different.

[0277] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-TRAILR2 VHH)2- first linker - Fc region- second linker - anti-CDH17 VHH. The two anti-TRAILR2 VHHs may be optionally operably linked to each another via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different. The two anti-TRAILR2 VHHs may bind to the same epitope on TRAILR2 or different epitopes on TRAILR2.

[0278] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: anti-CDH17 VHH - first linker - Fc region- second linker - (anti-TRAILR2 VHH)2. The two anti-TRAILR2 VHHs may be optionally operably linked to each another via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different. The two anti-TRAILR2 VHHs may bind to the same epitope on TRAILR2 or different epitopes on TRAILR2.

[0279] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)2- first linker - Fc region- second linker - (anti-TRAILR2 VHH)2. The two anti-CDH17 VHHs may be optionally operably linked to each another via a linker. The two anti-TRAILR2 VHHs may be optionally operably linked to each another via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different. The two anti-CDH17 VHHs may bind to the same epitope on CDH17 or different epitopes on CDH17. The two anti-TRAILR2 VHHs may bind to the same epitope on TRAILR2 or different epitopes on TRAILR2.66323195900v2Attorney Docket No: 260525.000128

[0280] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: (anti-CDH17 VHH)2 - first linker - Fc region- second linker - anti-TRAILR2 VHH. The two anti-CDH17 VHHs may be optionally operably linked to each another via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different. The two anti-CDH17 VHHs may bind to the same epitope on CDH17 or different epitopes on CDH17.

[0281] In a specific embodiment, a multispecific antigen-binding protein of the present disclosure may comprise two polypeptide chains, each polypeptide chain having the following structure from N-terminus to C-terminus: anti-CDH17 VHH - anti-TRAILR2 VHH - first linker - Fc region- second linker -anti-TRAILR2 VHH. The adjacent anti-TRAILR2 VHH and anti-CDH17 VHH may be optionally operably linked to one another via a linker. The multiple linkers used in the multispecific antigen-binding protein are not necessarily the same or different. The two anti-TRAILR2 VHHs may bind to the same epitope on TRAILR2 or different epitopes on TRAILR2.

[0282] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise a first and a second polypeptide chains, wherein the first polypeptide chain and / or the second polypeptide chain each comprises two antigen-binding domains (ABD1-ABD2 or ABD3-ABD4), three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), or four antigen-binding domains (ABD1-ABD4 or ABD5-ABD8).

[0283] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise a first and a second polypeptide chains, wherein the first polypeptide chain and the second polypeptide chain each comprise two antigen-binding domains (ABD1-ABD2 or ABD3-ABD4), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, and (ii) a second antigen-binding domain (ABD2) operably linked to the C-terminus of the first immunoglobulin Fc region; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a third antigen binding domain (ABD3) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, and (ii) a fourth antigen-binding domain (ABD4) operably linked to the C-terminus of the second immunoglobulin Fc region.

[0284] In some embodiments, ABDI and ABD3 specifically binds to CDH17, and ABD2 and ABD4 specifically bind to TRAILR2. In some embodiments, ABDI and ABD3 specifically bind to TRAILR2, and ABD2 and ABD4 specifically bind to CDH17.67323195900v2Attorney Docket No: 260525.000128

[0285] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise a first and a second polypeptide chains, wherein the first polypeptide chain and the second polypeptide chain each comprise three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, (ii) a second antigen-binding domain (ABD2) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker, and (iii) a third antigen-binding domain (ABD3) operably linked to the second antigen-binding domain (ABD2) via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fourth antigen binding domain (ABD4) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, (ii) a fifth antigen-binding domain (ABD5) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker, and (iii) a sixth antigen-binding domain (ABD6) operably linked to the fifth antigen-binding domain (ABD5) via a linker.

[0286] In some embodiments, ABDI and ABD4 specifically bind to CDH17, and ABD2, ABD3, ABD5, and ABD6 specifically bind to TRAILR2. In some embodiments, ABDI and ABD4 specifically bind to TRAILR2, and ABD2, ABD3, ABD5, and ABD6 specifically bind to CDH17.

[0287] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise a first and a second polypeptide chains, wherein the first polypeptide chain and the second polypeptide chain each comprise three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to a second antigen-binding domain (ABD2) via a linker, (ii) said second antigen binding domain (ABD2) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, and (iii) a third antigen-binding domain (ABD3) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fourth antigen binding domain (ABD4) operably linked to a fifth antigen-binding domain (ABD5) via a linker, (ii) said fifth antigen binding domain (ABD5) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, and (iii) a sixth antigen-binding domain (ABD6) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker.

[0288] In some embodiments, wherein ABDI, ABD2, ABD4, and ABD5 specifically bind to CDH17, and ABD3 and ABD6 specifically bind to TRAILR2. In some embodiments, ABDI, ABD2, ABD4, and ABD5 specifically bind to TRAILR2, and ABD3 and ABD6 specifically bind to CDH17.68323195900v2Attorney Docket No: 260525.000128

[0289] In some embodiments, the multispecific antigen-binding protein comprising a first and a second polypeptide chains, wherein the first polypeptide chain and the second polypeptide chain each comprise two antigen-binding domains (ABD1-ABD2 or ABD3-ABD4) is configured symmetrically. Nonlimiting examples of symmetric configurations include a multispecific antigen-binding protein in which antigen-binding domains ABDI and ABD3 are identical, and ABD2 and ABD4 are identical. Alternatively, each of ABDI, ABD2, ABD3, and ABD4 may be different.

[0290] In some embodiments, a multispecific antigen-binding protein of the present disclosure may comprise a first and a second polypeptide chains, wherein the first polypeptide chain and the second polypeptide chain each comprise four antigen-binding domains (ABD1-ABD4 or ABD5-ABD8), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to a second antigen-binding domain (ABD2) via a linker, ii) said second antigen binding domain (ABD2) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, (iii) a third antigen-binding domain (ABD3) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker, and iv) said third antigen-binding domain (ABD3) operably linked to a fourth antigen-binding domain (ABD4) via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fifth antigen binding domain (ABD5) operably linked to a sixth antigen-binding domain (ABD6) via a linker, ii) said sixth antigen binding domain (ABD6) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, (iii) a seventh antigen-binding domain (ABD7) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker, and iv) said seventh antigen-binding domain (ABD7) operably linked to an eighth antigen-binding domain (ABD8) via a linker.

[0291] In some embodiments, ABDI, ABD2, ABD5, and ABD6 specifically bind to CDH17, and ABD3, ABD4, ABD7, and ABD8 specifically bind to TRAILR2. In some embodiments, ABDI, ABD2, ABD5, and ABD6 specifically bind to TRAILR2, and ABD3, ABD4, ABD7, and ABD8 specifically bind to CDH17.

[0292] In some embodiments, the multispecific antigen-binding protein described herein is configured symmetrically. Non-limiting example of symmetric configuration includes a multispecific antigen-binding protein in which ABDI and ABD5 are identical, ABD2 and ABD6 are identical, ABD3 and ABD7 are identical, and ABD4 and ABD8 are identical.

[0293] In some embodiments, the multispecific antigen-binding protein described herein is configured asymmetrically. Non-limiting examples of asymmetric configuration include:i. A multispecific antigen-binding protein in which ABDI and ABD6 are identical, ABD2 and ABD5 are identical, ABD3 and ABD4 are identical, and ABD7 and ABD8 are identical; and69323195900v2Attorney Docket No: 260525.000128ii. A multispecific antigen-binding protein in which ABDI and ABD2 are identical, ABD3 and ABD4 are identical, ABD5 and ABD6 are identical, and ABD7 and ABD8 are identical.

[0294] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains (e.g., VHHs) and one or more anti CDH17 antigen-binding domains (e.g., VHHs) may be operably linked to one another in tandem. The adjacent antigen-binding domains (e.g., VHHs) may be optionally operably linked to one another via a linker. The linkers may be a flexible linker or a rigid linker.

[0295] The multispecific antigen-binding protein may also adopt any of the alternative molecular formats shown in Figure 30F and / or described in C. Spiess et al. Molecular Immunology 67 (2015) 95-106, which is incorporated herein by reference in its entirety.

[0296] Provided herein are multispecific antigen-binding proteins (e.g., antibodies such as singledomain antibodies) comprising a set of three CDRs ( / .e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0297] Provided herein are multispecific antigen-binding proteins (e.g., antibodies such as singledomain antibodies) comprising a set of three CDRs ( / . e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-CDH17 VHH antibodies listed in Table 1-3, Table 1-4, or Table 13, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0298] Provided herein are multispecific antigen-binding proteins (e.g., antibodies such as singledomain antibodies) comprising a set of three CDRs ( / . e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity and a set of three CDRs ( / .e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-CDH17 VHH antibodies listed in Table 1-3, Table 1-4, or Table 13, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0299] Provided herein are multispecific antigen-binding proteins (e.g., antibodies such as singledomain antibodies) comprising anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0300] Provided herein are multispecific antigen-binding proteins (e.g., antibodies such as singledomain antibodies) comprising anti-CDH17 VHH antibodies listed in Table 1-3, Table 1-4, or Table 13, or a similar sequence thereof having at least at least 70%, at least 75%, at least 80%, at least 85%, at least70323195900v2Attorney Docket No: 260525.00012890%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0301] Provided herein are multispecific antigen-binding proteins [e.g., antibodies such as singledomain antibodies) comprising anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity, and anti-CDH17 VHH antibodies listed in Table 1-3, Table 1-4, or Table 13, or a similar sequence thereof having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0302] In some embodiments, a multispecific antigen-binding protein described herein comprises an amino acid sequence selected from SEQ ID NOs: 4500-4506, 4509-4524, and 4526-4650, or a similar sequence thereof having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0303] In some embodiments, a multispecific antigen-binding protein described herein comprises an amino acid sequence selected from SEQ ID NOs: 4500-4506, 4509-4524, and 4526-4584, or a similar sequence thereof having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0304] In some embodiments, a multispecific antigen-binding protein described herein comprises an amino acid sequence selected from SEQ ID NOs: 4585-4650 or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0305] In some embodiments, a multispecific antigen-binding protein described herein comprises an amino acid sequence selected from SEQ ID NOs: 4587-4589, 4603, 4625, 4626, 4633, 4641, 4643, and 4644, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0306] In some embodiments, a multispecific antigen-binding protein described herein comprises an amino acid sequence SEQ ID NO: 4626, or a similar sequence thereof having at least at least 70%, at71323195900v2Attorney Docket No: 260525.000128least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0307] In some embodiments, a multispecific antigen-binding protein described herein has CDH17-dependent agonistic activity.Antigen-binding Domains

[0308] The antigen-binding domains of the present disclosure can include an antibody or an antigenbinding fragment of an antibody, such as a human antibody, a humanized antibody; a camelid antibody; a chimeric antibody; a recombinant antibody; a heavy chain antibody; a single-domain antibody (e.g., VHH); a single chain antibody (e.g., single chain fragment variable (scFv)); a diabody; a triabody; a tetrabody; a Fab fragment; a F(ab') 2 fragment; an IgD antibody; an IgE antibody; an IgM antibody; an IgGl antibody; an lgG2 antibody; an lgG3 antibody; or an lgG4 antibody, and fragments thereof.

[0309] In some embodiments, an antigen-binding domain that binds to TRAILR2 or CDH17 is a singledomain antibody (also termed as "sdAb"). The single-domain antibodies of the present disclosure can be derived from numerous sources, including but not limited to VHHs, VNARs, or VH domains (naturally occurring or engineered VH domains). VHHs can be generated from camelid heavy chain only antibodies and libraries thereof. VNARs can be generated from cartilaginous fish heavy chain only antibodies and libraries thereof. Various methods have been implemented to generate monomeric sdAbs from conventionally heterodimeric VH and VL domains, including interface engineering and selection of specific germline families. In some embodiments, the sdAbs of the present invention are human or humanized.

[0310] In some embodiments, a single-domain antibody described herein is a VHH fragment (also known as a nanobody). VHH fragments are also referred to as " V-bodies" in the present disclosure. In some embodiments, the VHH is a camelid VHH, a humanized VHH or a camelized VH. In some embodiments, a single-domain antibody described herein is a VH domain. In some embodiments, a single-domain antibody described herein is a naturally occurring VH domain or engineered VH domain.

[0311] The variable domain of an antigen-binding protein (e.g., antibody such as a single-domain antibody) of the present disclosure comprises at least three complementarity determining regions (CDRs) which determine its binding specificity. Preferably, in a variable domain, the CDRs are distributed between framework regions (FRs). The variable domain typically contains 4 framework regions interspaced by 3 CDR regions, resulting in the following typical antibody variable domain structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. CDRs and / or FRs of the single-domain antibody of the present72323195900v2Attorney Docket No: 260525.000128disclosure may be fragments or derivatives from a naturally occurring antibody variable domain or may be synthetic.

[0312] Binding affinity of a molecular interaction between two molecules, e.g., for an antigen recognized by a multispecific antigen as described herein, can be measured via various techniques, such as surface plasmon resonance (SPR), bio-layer interferometry (BLI), enzyme-linked immunosorbent assay (ELISA), equilibrium dialysis, fluorescent-activated cell sorting (FACS), or flow cytometry binding assays and the like. Surface plasmon resonance is a biosensor technique that allows for the analysis of real-time biospecific interactions by detection of alterations in protein concentrations within a biosensor matrix, where one molecule is immobilized on the biosensor chip and the other molecule is passed over the immobilized molecule under flow conditions (see e.g., Ober et al. 2001, Intern. Immunology 13: 1551-1559). SPR can for example be performed using the BIACORE® system or Carterra LSA system. Another biosensor technique that can be used to determine affinities of biomolecular interactions is biolayer interferometry (BLI) (see e.g., Abdiche et al. 2008, Anal. Biochem. 377: 209-217). Bio-layer Interferometry is a label-free optical technique that analyzes the interference pattern of light reflected from two surfaces: an internal reference layer (reference beam) and a layer of immobilized protein on the biosensor tip (signal beam). A change in the number of molecules bound to the tip of the biosensor causes a shift in the interference pattern, reported as a wavelength shift (nm), the magnitude of which is a direct measure of the number of molecules bound to the biosensor tip surface. Since the interactions can be measured in real-time, association and dissociation rates and affinities can be determined. BLI can for example be performed using the Octet® Systems. Alternatively, affinities can be measured in Kinetic Exclusion Assay (KinExA) (see e.g., Drake et al. 2004, Anal. Biochem., 328: 35-43), which is a solution-based method to measure true equilibrium binding affinity and kinetics of unmodified molecules. Equilibrated solutions of an antibody / antigen complex are passed over a column with beads precoated with antigen (or antibody), allowing the free antibody (or antigen) to bind to the coated molecule. Detection of the antibody (or antigen) thus captured is accomplished with a fluorescently labeled protein binding the antibody (or antigen).Anti-TRAILR2 Antigen-binding Domains

[0313] The present disclosure provides antigen-binding domains (e.g., antibodies, such as singledomain antibodies) that bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2, also known as Death Receptor 5 or DR5).73323195900v2Attorney Docket No: 260525.000128

[0314] TRAIL receptor 2 (TRAILR2) is a single pass type-1 membrane protein with a molecular weight of ~42 kDa that belongs to the TNFR superfamily. TRAILR2 is comprised of an extracellular domain with three cysteine-rich domains (CRDs) and an intracellular death domain that is involved in apoptotic signaling. The cysteine-rich domains contain a total of 7 disulfide bonds stabilizing the elongated structure of the protein. CRD1 is also called pre-ligand assembly domain (PLAD) and has been found to be involved in low affinity (>1 pM) TRAILR2 dimer formation in the absence of TRAIL. The TRAILR2 death domain is a bundle of six alpha helices that are involved in the binding of intracellular binding partners and formation of the death-inducing signaling complex (DISC) to promote apoptotic signaling. Further, TRAILR2 has been found to be O-glycosylated at two positions within CRD1 and CRD2 altering its ability to induce apoptosis albeit not changing the affinity towards TRAIL. In addition, fucosylation of the O-glycans has been reported to be important for apoptosis induction capabilities of TRAILR2.

[0315] The TRAIL ligand itself consists of a -sandwich fold and homo-trimerizes in solution. The trimer is non-covalent, and TRAIL dissociates at low concentration leading to the fast degradation of the monomeric units. The TRAIL trimer is stabilized by a zinc ion that is coordinated by 3 free cysteines (one per monomer) at the trimerization interface. The TRAIL ligand exists both in a soluble (sTRAIL) and membrane-anchored (mTRAIL) version.

[0316] The extracellular domain of TRAILR2 has been structurally characterized as a trimeric complex in presence of the TRAIL ligand. In addition, an NMR structure of the transmembrane domain trimer and hexamer has been reported, while there are no structural data available on the intracellular death domain of TRAILR2. However, homolog structures of death domains and further signaling components provide structural insights into the signaling complex.

[0317] In the absence of the trimeric TRAIL ligand, TRAILR2 has been reported to exist either in monomeric or dimeric state. The dimeric state is mediated via a low affinity (>1 pM) interaction of the PLAD domains. The dimers have been reported to adopt two different conformations, an active, parallel conformation in which the TRAIL binding site is accessible or an inactive, anti-parallel conformation, in which the TRAIL binding site is shielded. In the presence of soluble or membrane bound trimeric TRAIL, TRAILR2 can bind to TRAIL trimers with high affinity (<2 nM) and assemble into a hexameric TRAILR23TRAIL3complex with 3 TRAILR2 molecules bound to trimeric TRAIL on the membrane. The TRAIL binding site on TRAILR2 is located at the concave surface formed by CRD1 and CRD2. Furthermore, it has been reported that in addition to homogenous complexes formed by TRAILR2 bound to trimeric TRAIL, heterogenous complexes with TRAILR1 and TRAILR4 can be assembled exhibiting an overall lower signaling potency when activated. As for other receptors of the TNFR-superfamily it has also been74323195900v2Attorney Docket No: 260525.000128reported for TRAILR2 that dimers and trimers may not be randomly distributed throughout the membrane but may be organized in a higher order hexagonal fashion.

[0318] The actual distribution of TRAILR2 oligomeric states on any given normal or cancer cell remains largely unknown. It has been confirmed thatTNFRl mainly exists in a monomeric and dimeric state in the absence of the TNF ligand, while in the presence of TNF, most of the receptors assemble into trimers or higher oligomers of the trimers while only few monomers and no dimers were observed. Whether these results can be translated to TRAILR2 remains to be seen.

[0319] In line with these observations, it has been consistently reported that TRAILR2 oligomerization may be needed for efficient signaling. The precise mechanism of signaling complex assembly and protein stoichiometries are not yet fully understood, but it is assumed that two TRAIL bound TRAILR2 trimers need to come into close proximity to enable efficient apoptotic signaling. In more detail, membranebound TRAIL trimers were shown to bind three molecules of TRAILR2 receptors that assemble in a symmetrical trimeric complex, with each TRAILR2 molecule interacting with two TRAIL monomers in the center of the complex. This trimerization has been suggested to induce a rearrangement of the transmembrane helices, resulting in a conformational change of the TRAILR2 death domains into an open conformation. In its open conformation the TRAILR2-DD can recruit an adaptor protein called Fas associated death domain (FADD) via a tight helix-helix interaction similar to Fas-FADD. Once this interaction is established, procaspase-8 can bind to the now exposed FADD death-effector domain (DED). A single death-inducing signaling complex (DISC) complex was found to be unable to induce downstream signaling as procaspase-8 requires homodimerization to exhibit autocatalytic activity and subsequent activation. Hence, a second TRAILR2 complex with all intracellular components bound may need to be in close proximity to enable procaspase-8 dimerization. DISC complex dimerization was mediated only via membrane bound (mTRAIL) but not soluble TRAIL. Anchoring of mTRAIL in the lipid bilayer restricts diffusion of the protein to two dimensions, which would effectively increase the colocalization of mTRAIL molecules, and in turn increase the efficiency of agonist-induced TRAILR2 clustering compared to that promoted by the soluble form.

[0320] A structural model proposed for the entire signaling complex indicates that a distance of >90A is needed between two TRAILR23TRAIL3complexes to accommodate the intracellular DISC complex. This distance correlates well with the distance spanned by monoclonal antibodies (mAbs), which have been shown to induce TRAILR2 clustering and signaling.

[0321] In some embodiments, a human TRAILR2 can be encoded by a TNF receptor superfamily member 10b (TNFRSF10B) gene (NCBI Gene ID 8795) and has the amino acid sequence75323195900v2Attorney Docket No: 260525.000128MEQRGQNAPAASGARKRHGPGPREARGARPGPRVPKTLVLVVAAVLLLVSAESALITQQDLAPQQRAAPQQKRSSPS EGLCPPGHHISEDGRDCISCKYGQDYSTHWNDLLFCLRCTRCDSGEVELSPCTTTRNTVCQCEEGTFREEDSPEMCRKC RTGCPRGMVKVGDCTPWSDIECVHKESGTKHSGEVPAVEETVTSSPGTPASPCSLSGIIIGVTVAAVVLIVAVFVCKSLL WKKVLPYLKGICSGGGGDPERVDRSSQRPGAEDNVLNEIVSILQPTQVPEQEMEVQEPAEPTGVNMLSPGESEHLLEP AEAERSQRRRLLVPANEGDPTETLRQCFDDFADLVPFDSWEPLMRKLGLMDNEIKVAKAEAAGHRDTLYTMLIKWVN KTGRDASVHTLLDALETLGERLAKQKIEDHLLSSGKFMYLEGNADSAMS (UniProtKB Accession No. 014763) (SEQ ID NO: 2149).

[0322] In some embodiments, a cyno TRAILR2 can be encoded by a TNF receptor superfamily member 10b (TNFRSF10B) gene (NCBI Gene ID 102133727) and has the amino acid sequence MGQLRQSAPAASGARKGRGPGPREARGARPGLRVLKTLVLVVAAAAVLLSVSADCAPITRQSLDPQRRAAPQQKRSSP TEGLCPPGHHISEDSRECISCKYGQDYSTHWNDFLFCLRCTKCDSGEVEVNSCTTTRNTVCQCEEGTFREEDSPEICRKC RTGCPRGMVKVKDCTPWSDIECVHKESGTKHTGEVPAVEKTVTTSPGTPASPCSLSGIIIGVIVLVVIVVVAVIVWKTSL WKKVLPYLKGVCSGGGGDPERVDSSSHSPQRPGAEDNALNEIVSIVQPSQVPEQEMEVQEPAEQTDVNTLSPGESEHL LEPAKAEGPQRRGQLVPVNENDPTETLRQCFDDFAAIVPFDAWEPLVRQLGLTNNEIKVAKAEAASSRDTLYVMLIKW VNKTGRAASVNTLLDALETLEERLAKQKIQDRLLSSGKFMYLEDNADSATS (UniProtKB Accession No.A0A2K5TXK0 | A0A2K5TXK0) (SEQ ID NO: 2150).

[0323] In some embodiments, a mouse TRAILR2 can be encoded by a TNF receptor superfamily member 10b (TNFRSF10B) gene (NCBI Gene ID 21933) and has the amino acid sequence MEPPGPSTPTASAAARADHYTPGLRPLPKRRLLYSFALLLAVLQAVFVPVTANPAHNRPAGLQRPEESPSRGPCLAGQY LSEGNCKPCREGIDYTSHSNHSLDSCILCTVCKEDKVVETRCNITTNTVCRCKPGTFEDKDSPEICQSCSNCTDGEEELTSC TPRENRKCVSKTAWASWHKLGLWIGLLVPVVLLIGALLVWKTGAWRQWLLCIKRGCERDPESANSVHSSLLDRQTSST TNDSNHNTEPGKTQKTGKKLLVPVNGNDSADDLKFIFEYCSDIVPFDSWNRLMRQLGLTDNQIQMVKAETLVTREALY QMLLKWRHQTGRSASINHLLDALEAVEERDAMEKIEDYAVKSGRFTYQNAAAQPETGPGGSQCV (UniProtKB Accession No. Q9QZM4) (SEQ ID NO: 2151).

[0324] In various embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure have an agonist effect upon binding to TRAILR2. While not wishing to be bound by theory, an agonistic TRAILR2 binder can promote or increase activation of TRAILR2 and / or potentiate one or more signal transduction pathways mediated by TRAILR2. Agonistic TRAILR2 binders may promote or increase TRAILR2 activation by binding TRAILR2, e.g., to induce TRAILR2 multimerization which can render the receptor biologically active. For example, agonistic TRAILR2 binders may nucleate the trimerization of TRAILR2 and may additionally act to bring a TRAILR2 agonistic binder-associated TRAILR2 trimer into close proximity with one or more additional TRAILR2 agonistic binder-associated TRAILR2 trimer(s) in a76323195900v2Attorney Docket No: 260525.000128manner similar to that which occurs when TRAILR2 interacts with its cognate ligand, TRAIL, thus inducing TRAILR2-mediated signaling. In some embodiments, an agonistic TRAILR2 binder described herein may bind, e.g., three molecules of TRAILR2 which can assemble into a trimeric complex ( / .e., a TRAILR2 trimer).

[0325] In some embodiments, agonistic TRAILR2 binding domains of the present disclosure may be capable of rendering a TRAILR2 described herein capable of binding intracellular binding partners (e.g., adaptor proteins such as but not limited to Fas associated death domain (FADD)) and forming a deathinducing signaling complex (DISC) to promote apoptotic signaling.

[0326] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind to human TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to human TRAILR2 with a KDof less than about lxlO-6M, for example, less than about 5xl0-7M, less than about 3xl0-7M, less than about lxlO-7M, less than about 8xl0-8M, less than about 5xl0-8M, less than about 3xl0-8M, less than about lxlO-8M, less than about 9xl0-9M, less than about 8xl0-9M, less than about 7xl0-9M, less than about 6xl0“9M, less than about 5xl0-9M, less than about 4xl0-9M, less than about 3xl0-9M, less than about 2xl0“9M, or less than about lxlO-9M, or about lxlO-10to lxlO-9M, about lxlO-10to 5xl0-9M, about lxlO-10to lxlO-8M, about lxlO-10to 5xl0-8M, about lxlO-9to 5xl0-9M, about 5xl0-9to lxlO-8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8M, about lxlO-9to lxlO-7M, or about lxlO-8to lxlO-7M.

[0327] In some embodiments, anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human TRAILR2 with a KDof less than about 3xl0-7M. In some embodiments, anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human TRAILR2 with a KDof about lxlO-10to 5xl0-8M.

[0328] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind to cynomolgus monkey ("cyno") TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to cyno TRAILR2 with a KDof less than about lxlO-6M, for example, less than about 5xl0-7M, less than about 3xl0-7M, less than about lxlO-7M, less than about 8xl0-8M, less than about 5xl0-8M, less than about 3xl0-8M, less than about lxlO-8M, less than about 8xl0-9M, less than about 5xl0-9M, less than about 3xl0-9M, or less than about lxlO-9M, or about lxlO-10to lxlO-9M, about lxlO-10to 5xl0-9M, about lxlO-10to lxlO-8M, about lxlO-10to 5xl0-8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8M, about lxlO-977323195900v2Attorney Docket No: 260525.000128to 1x10-7M, about 1x10-9to 2x10-7M, about 1x10-9to 5x10-7M, about 1x10-8to 1x10-7M, about lxlO-8to 2xl0“7M, about lxlO-8to 5xl0-7M, or about lxlO-8to lxlO-6M.

[0329] In some embodiments, anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to cyno TRAILR2 with a KDof less than about 3xl0“7M. In some embodiments, anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to cyno TRAILR2 with a KDof about lxlO-9to IxlO’7M.

[0330] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind to mouse TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure may bind to mouse TRAILR2 with a KDof less than about 1x10-6M, for example, less than about 5xl0“7M, less than about 3xl0-7M, less than about lxlO-7M, less than about 8xl0-8M, less than about 5xl0“8M, less than about 3xl0-8M, less than about lxlO-8M, less than about 8xl0-9M, less than about 5xl0-9M, less than about 3xl0-9M, or less than about lxlO-9M, or about lxlO-10to lxlO-9M, about lxlO-10to 5xl0-9M, about lxlO-10to lxlO-8M, about lxlO-10to 5xl0-8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8M, about lxlO-9to lxlO-7M, about lxlO-9to 2xl0-7M, about lxlO-9to 5xl0-7M, about lxlO-8to lxlO-7M, about lxlO-8to 2xl0-7M, about lxlO-8to 5xl0-7M, or about lxlO-8to lxlO-6M. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure do not bind to mouse TRAILR2.

[0331] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind to rat TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure may bind to rat TRAILR2 with a KDof less than about lxlO-6M, for example, less than about 5xl0-7M, less than about 3xl0-7M, less than about lxlO-7M, less than about 8xl0-8M, less than about 5xl0-8M, less than about 3xl0-8M, less than about lxlO-8M, less than about 8xl0-9M, less than about 5xl0-9M, less than about 3xl0-9M, or less than about lxlO-9M, or about lxlO-10to lxlO-9M, about lxlO-10to 5xl0-9M, about lxlO-10to lxlO-8M, about lxlO-10to 5xl0-8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8M, about lxlO-9to lxlO-7M, about lxlO-9to 2xl0-7M, about lxlO-9to 5xl0-7M, about lxlO-8to lxlO-7M, about lxlO-8to 2xl0-7M, about lxlO-8to 5xl0-7M, or about lxlO-8to lxlO-6M. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure do not bind to rat TRAILR2.

[0332] In some embodiments, anti-TRAILR2 antigen-binding domains described herein do not block binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2.78323195900v2Attorney Docket No: 260525.000128

[0333] In some embodiments, anti-TRAILR2 antigen-binding domains described herein blocks binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2. In some embodiments, the anti-TRAILR2 antigen-binding domains described herein may block binding of TNF-related apoptosis-inducing ligand (TRAIL) toTRAILR2 by from about 10% or more. In some embodiments, the binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2 can be blocked by the anti-TRAILR2 antigen-binding domains described herein by from about 10% to about 20%, from about 10% to about 30%, from about 10% to about 40%, from about 10% to about 50%, from about 10% to about 60%, from about 10% to about 70%, from about 10% to about 80%, from about 10% to about 90%, more than 60%, from about 60% to about 70%, from about 60% to about 80%, from about 60% to about 90%, more than about 70%, from about 70% to about 80%, from about 70% to about 90%, more than about 80%, from about 80% to about 90%, more than 90%, from about 90% to about 95%, from about 90% to about 98%, more than 95%, from about 95% to about 98%, more than about 98%, or more than about 99%. The binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2 can be blocked by the anti-TRAILR2 antigenbinding domains described herein by about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or even 100%. In some embodiments, the binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2 can be blocked by the anti-TRAILR2 antigen-binding domains described herein by about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, or more.

[0334] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure may specifically bind TRAILR2 without exhibiting specific binding for another receptor of the tumor necrosis factor receptor (TNFR) superfamily.

[0335] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure do not bind specifically to TRAILR1, TRAILR3 and / or TRAILR4. In some embodiments, anti-TRAILR2 antigenbinding domains of the present disclosure do not bind specifically to TRAILR1. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure do not bind specifically to TRAILR3. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure do not bind specifically to TRAILR4. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure do not bind specifically to TRAILR1, TRAILR3 and TRAILR4.

[0336] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind cysteine-rich domain (CRD) domain 1 (CRD1) and / or CRD2 of TRAILR2. In some embodiments, anti-79323195900v2Attorney Docket No: 260525.000128TRAILR2 antigen-binding domains of the present disclosure bind CRD1 of TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind CRD2 of TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind CRD1 and CRD2 ofTRAILR2.

[0337] In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind CRD2 and / or CRD3 of TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind CRD2 of TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind CRD3 of TRAILR2. In some embodiments, anti-TRAILR2 antigen-binding domains of the present disclosure bind CRD2 and CRD3 of TRAILR2.

[0338] Binding affinity of a molecular interaction between two molecules can be measured via various techniques, such as surface plasmon resonance (SPR), bio-layer interferometry (BLI), enzyme-linked immunosorbent assay (ELISA), equilibrium dialysis, fluorescent-activated cell sorting (FACS), flow cytometry binding assays, or isothermal titration calorimetry (ITC), and the like. Surface plasmon resonance is a biosensor technique that allows for the analysis of real-time biospecific interactions by detection of alterations in protein concentrations within a biosensor matrix, where one molecule is immobilized on the biosensor chip and the other molecule is passed over the immobilized molecule underflow conditions (see e.g., Ober et al. 2001, Intern. Immunology 13: 1551-1559). SPR can for example be performed using the BIACORE® system or Carterra LSA system. Another biosensor technique that can be used to determine affinities of biomolecular interactions is bio-layer interferometry (BLI) (see e.g., Abdiche et al. 2008, Anal. Biochem. 377: 209-217). Bio-layer Interferometry is a label-free optical technique that analyzes the interference pattern of light reflected from two surfaces: an internal reference layer (reference beam) and a layer of immobilized protein on the biosensor tip (signal beam). A change in the number of molecules bound to the tip of the biosensor causes a shift in the interference pattern, reported as a wavelength shift (nm), the magnitude of which is a direct measure of the number of molecules bound to the biosensor tip surface. Since the interactions can be measured in real-time, association and dissociation rates and affinities can be determined. BLI can for example be performed using the Octet® Systems. Alternatively, affinities can be measured in Kinetic Exclusion Assay (KinExA) (see e.g., Drake et al. 2004, Anal. Biochem., 328: 35-43), which is a solution-based method to measure true equilibrium binding affinity and kinetics of unmodified molecules. Equilibrated solutions of an antibody / antigen complex are passed over a column with beads precoated with antigen (or antibody), allowing the free antibody (or antigen) to bind to the coated molecule. Detection of the antibody (or80323195900v2Attorney Docket No: 260525.000128antigen) thus captured is accomplished with a fluorescently labeled protein binding the antibody (or antigen).

[0339] Anti-TRAILR2 antigen-binding domains of the present disclosure can include an antibody or an antigen-binding fragment of an antibody, such as a human antibody, a humanized antibody; a camelid antibody; a chimeric antibody; a recombinant antibody; a heavy chain antibody; a single-domain antibody {e.g., VHH); a single chain antibody {e.g., single chain fragment variable (scFv)); a diabody; a triabody; a tetrabody; a Fab fragment; a F(ab') 2 fragment; an IgD antibody; an IgE antibody; an IgM antibody; an IgGl antibody; an lgG2 antibody; an lgG3 antibody; or an lgG4 antibody, and fragments thereof.

[0340] In some embodiments, an anti-TRAILR2 antigen-binding domain that binds to TRAILR2 is a single-domain antibody (also termed as "sdAb"). The single-domain antibodies of the present disclosure can be derived from numerous sources, including but not limited to VHHs, VNARs, or VH domains (naturally occurring or engineered VH domains). VHHs can be generated from camelid heavy chain only antibodies and libraries (e.g., synthetic libraries) thereof. VNARs can be generated from cartilaginous fish heavy chain only antibodies and libraries (e.g., synthetic libraries) thereof. Various methods have been implemented to generate monomeric sdAbs from conventionally heterodimeric VH and VL domains, including interface engineering and selection of specific germline families. In some embodiments, the sdAb of the present invention are human or humanized.

[0341] In some embodiments, an anti-TRAILR2 single-domain antibody described herein is a VHH fragment (also known as a nanobody). VHH fragments are also referred to as " V-bodies" in the present disclosure. In some embodiments, the VHH is a camelid VHH, a humanized VHH or a camelized VH. In some embodiments, a single-domain antibody described herein is a VH domain. In some embodiments, a single-domain antibody described herein is a naturally occurring VH domain or engineered VH domain.

[0342] The variable domain of an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises at least three complementarity determining regions (CDRs) which determine its binding specificity. Preferably, in a variable domain, the CDRs are distributed between framework regions (FRs). The variable domain typically contains 4 framework regions interspaced by 3 CDR regions, resulting in the following typical antibody variable domain structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. CDRs and / or FRs of the single domain antibody of the invention may be fragments or derivatives from a naturally occurring antibody variable domain or may be synthetic.81323195900v2Attorney Docket No: 260525.000128

[0343] Sequence identifiers corresponding to exemplary anti-TRAILR2 VHH antibodies provided herein are listed in Table 1-1. Table 1-1 sets forth the sequence identifiers of amino acid sequences of the complementarity determining regions (CDR1, CDR2 and CDR3), amino acid and DNA sequences of the full-length camelid VHH antibodies, as well as amino acid sequences of corresponding humanized VHH antibodies. In the present disclosure, the suffix "-Hui" in an antibody ID indicates a humanized version of the referenced antibody. Amino acid sequences of additional exemplary anti-TRAILR2 VHH antibodies and corresponding humanized VHH antibodies are provided in Table 1-2.Table 1-1. Sequence identifiers for exemplary anti-TRAILR2 VHH antibodiesNon-humanized Humanized CDR1 CDR2 CDR3VHH VHHAntibody ID GroupDNA Amino acid Amino acid sequencesequence sequence T00001 A 1 2 3 4 57 5 T00002 B 6 7 8 9 58 10 T00003 C 11 12 13 14 59 15 T00004 C 11 12 16 17 60 18 T00005 C 11 19 20 21 61 22 T00006 C 23 24 25 26 131 27 T00007 D 28 29 30 31 132 32 T00007Hul Glyl D 28 29 2298 - - 2354 T00007Hul Gly2 D 28 29 2299 - - 2355 T00007HU1 Gly3 D 28 29 2300 - - 2356 T00007HU1 Ala4 D 28 29 2301 - - 2357 T00007Hul Ala5 D 28 29 2302 - - 2358 T00007Hul Ala6 D 28 29 2303 - - 2359 T00007HU1 Ala7 D 28 29 2304 - - 2360 T00007HU1 Ala8 D 28 29 2305 - - 2361 T00007Hul Ala9 D 28 29 2306 - - 2362 T00007Hul AlalO D 28 29 2307 - - 2363 T00007Hul Alall D 28 29 2308 - - 2364 T00007HU1 Alal2 D 28 29 2309 - - 2365 T00007HU1 Alal3 D 28 29 2310 - - 2366 T00007HU1 Alal4 D 28 29 2311 - - 2367 T00007Hul Alal5 D 28 29 2312 - - 2368 T00007HU1 Alal6 D 28 29 2313 - - 2369 T00007HU1 Alal7 D 28 29 2314 - - 2370 T00008 E 33 34 35 36 133 37 T00009 F 38 39 40 41 134 42T00010 G 43 39 44 45 135 4682323195900v2Attorney Docket No: 260525.000128Non-humanized Humanized CDR1 CDR2 CDR3VHH VHHAntibody ID GroupDNA Amino acid Amino acid sequencesequence sequence T00011 H 47 48 49 50 136 51 T00012 1 52 53 54 55 137 56 T00012HU1 Alai 1 52 53 2315 - - 2371 T00012HU1 Gly2 1 52 53 2316 - - 2372 T00012Hul Ala3 1 52 53 2317 - - 2373 T00012Hul Ala4 1 52 53 2318 - - 2374 T00012Hul Ala5 1 52 53 2319 - - 2375 T00012HU1 Ala6 1 52 53 2320 - - 2376 T00012HU1 Ala7 1 52 53 2321 - - 2377 T00012Hul Ala8 1 52 53 2322 - - 2378 T00012Hul Ala9 1 52 53 2323 - - 2379 T00012HU1 AlalO 1 52 53 2324 - - 2380 T00012HU1 Alall 1 52 53 2325 - - 2381 T00012Hul Alal2 1 52 53 2326 - - 2382 T00012Hul Alal3 1 52 53 2327 - - 2383 T00012Hul Alal4 1 52 53 2328 - - 2384 T00013 1 52 53 54 55 137 56 TOGO 14 J 62 63 64 65 138 66 T00015 J 67 68 69 70 139 71 T00016 K 72 73 74 75 140 76 T00017 K 72 77 78 79 141 80 T00018 L 81 82 83 84 142 85 T00019 M 86 87 88 89 143 90 T00020 N 28 91 92 93 144 94 T00021 0 95 96 97 98 145 99 T00022 P 2189 2190 2191 2192 2193 2194 T00022Hul Alai P 2189 2190 2329 - - 2385 T00022Hul Gly2 P 2189 2190 2330 - - 2386 T00022Hul Ala3 P 2189 2190 2331 - - 2387 T00022HU1 Ala4 P 2189 2190 2332 - - 2388 T00022HU1 Ala5 P 2189 2190 2333 - - 2389 T00022Hul Ala6 P 2189 2190 2334 - - 2390 T00022Hul Ala7 P 2189 2190 2335 - - 2391 T00022HU1 Ala8 P 2189 2190 2336 - - 2392 T00022HU1 Ala9 P 2189 2190 2337 - - 2393 T00023 Q 2195 2196 2197 2198 2199 2200 T00023Hul Alai Q 2195 2196 2338 - - 2394T00023Hul Ala2 Q 2195 2196 2339 - - 239583323195900v2Attorney Docket No: 260525.000128Non-humanized Humanized CDR1 CDR2 CDR3VHH VHHAntibody ID GroupDNA Amino acid Amino acid sequencesequence sequence T00023Hul Ala3 Q 2195 2196 2340 - - 2396 T00023Hul Ala4 Q 2195 2196 2341 - - 2397 T00023HU1 Ala5 Q 2195 2196 2342 - - 2398 T00023HU1 Ala6 Q 2195 2196 2343 - - 2399 T00023Hul Ala7 Q 2195 2196 2344 - - 2400 T00024 R 2201 2202 2203 2204 2205 2206 T00024Hul Alai R 2201 2202 2345 - - 2401 T00024HU1 Ala2 R 2201 2202 2346 - - 2402 T00024HU1 Gly3 R 2201 2202 2347 - - 2403T00024Hul Ala4 R 2201 2202 2348 - - 2404 Table 1-2. Sequence identifiers for additional exemplary anti-TRAILR2 VHH antibodies and humanized VHH antibodiesNon-humanized VHH Humanized VHHGroupAmino Acid SequenceA 4 5A 146 469B 9 10B 147 470B 148 471C 149 472C 150 473C 151 474C 152 475C 21 22C 153 476C 154 477C 155 472C 156 478c 157 479c 158 479c 159 480c 160 481c 161 482c 162 483c 163 484c 164 485c 165 486c 166 48784323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceC 167 488C 168 489c 169 490c 170 491c 171 492c 172 493c 173 494c 174 495c 175 496c 176 497c 177 498c 178 499c 179 500c 180 501c 181 472c 182 502c 183 503c 184 504c 185 505c 186 506c 187 507c 188 508c 189 509c 190 510c 191 511c 192 512c 193 513c 194 514c 195 515c 196 516c 197 517c 198 518c 199 519c 200 520c 201 521c 202 522c 203 523c 204 524c 205 525c 206 526c 207 527c 208 528c 209 52985323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceC 210 530C 211 531c 212 532c 213 533c 214 534c 215 483c 216 535c 217 536c 218 537c 219 538c 220 539c 221 540c 222 541c 223 542c 224 543c 225 544c 226 545c 227 546c 228 547c 229 548c 230 549c 231 550c 232 551c 233 552c 234 553c 235 554c 236 555c 237 556c 238 557c 239 476c 240 509c 241 558c 242 559c 243 560c 244 561c 245 562c 246 563c 247 564c 248 565c 249 566c 250 567c 251 568c 252 56986323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceC 253 570C 254 481c 255 571c 256 572c 257 573c 258 574c 259 575c 260 576c 261 577c 262 578c 263 579c 264 580c 265 581c 266 582c 267 583c 268 584c 269 585c 270 586c 271 587c 272 588c 273 589c 274 590c 275 591c 276 592c 277 593c 278 594c 279 595c 280 596c 281 597c 282 598c 283 599c 284 600c 285 601c 286 602c 287 603c 288 604c 289 605c 290 606c 291 607c 292 608c 293 609c 294 610c 295 61187323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceC 296 612C 297 613c 298 614c 299 615c 300 616c 301 617c 302 618c 303 619c 304 620c 305 621c 306 622c 307 623c 308 624c 309 625c 310 626c 311 627c 312 628c 313 629c 314 630c 315 631c 316 632c 317 633c 318 634c 319 635c 320 636c 321 637c 322 638c 323 639c 324 640c 325 641c 326 642c 327 643c 328 644c 329 645c 330 646c 331 647c 332 648c 333 649c 334 650c 335 651c 336 652c 337 653c 338 65488323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceC 339 655C 340 656c 341 657c 342 658c 343 659c 344 660c 345 661c 346 662c 347 663c 348 664c 349 665c 350 666c 351 667c 352 668c 353 669c 26 27c 354 670c 355 671c 356 672c 357 673c 358 674c 359 27c 360 675c 361 676c 14 15c 17 18D 362 677D 31 32D 363 678D 364 679D 365 680D 366 681D 367 682D 368 683D 369 684D 370 685D 371 686D 372 686D 373 687D 374 688D 375 689D 376 690D 377 69189323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceD 378 685D 379 692D 380 693D 381 694D 382 695D 383 696D 384 697D 385 698D 386 699D 387 700D 388 701D 389 702D 390 703D 391 704D 392 705D 393 706D 394 707D 395 708D 396 709D 397 710D 398 711D 399 712D 400 713D 401 714D 402 715D 403 716E 404 717E 405 718E 406 719E 407 720E 408 721E 409 722E 410 723E 411 724E 412 717E 413 725E 414 726E 415 717E 416 727E 417 717E 418 728E 36 37F 41 4290323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceF 419 729F 420 730F 421 42F 422 729F 423 729F 424 42F 425 731G 426 732G 427 733G 45 46G 428 734G 429 735G 430 736G 431 737H 50 51H 432 51H 433 511 55 561 434 7381 435 7391 436 7401 437 7411 438 7421 439 7431 440 7441 441 7451 442 7461 443 7471 444 7481 445 7491 446 7501 447 7511 448 7521 55 56J 70 71J 449 71J 65 66J 450 753K 75 76K 79 80K 451 76K 452 80K 453 7691323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceK 454 80K 455 754K 456 76K 457 755K 458 76L 84 85L 459 756L 460 757L 461 85M 89 90N 93 94N 462 7580 98 990 463 7590 464 7600 465 7600 466 7610 467 990 468 99P 2214 2234P 2192 2194Q 2215 2235Q 2198 2200Q 2216 2236Q 2217 2237Q 2218 2235Q 2219 2200R 2220 2238R 2221 2239R 2204 2206R 2222 2240R 2223 2241R 2224 2242R 2225 2243R 2226 2244R 2227 2245R 2228 2246R 2229 2247R 2230 2248R 2231 2249R 2232 2250R 2233 225192323195900v2Attorney Docket No: 260525.000128

[0344] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) described herein may further comprise a complementarity determining region 1 (CDR1) comprising an amino acid sequence selected from (amino acids listed in a pair of brackets represent the possible amino acids at the particular position):a). GRTFSSNL (SEQ ID NO: 1);b). GGT(F / L)(A / S)N(D / N)G (SEQ ID NO: 113);c). GRTL(D / N / S)(A / D / E)Y(A / G) (SEQ ID NO: 114);d). GRTFS(N / S)YA (SEQ ID NO: 115);e). GRDFSNYV (SEQ ID NO: 33);f). G(L / R)(I / S)FS(D / S)YA (SEQ ID NO: 116);g). GR(A / T)FSTLA (SEQ ID NO: 117);h). GRTFSSDI (SEQ ID NO: 47);i). GRSFGD(D / F / Y)A (SEQ ID NO: 118);j). G(G / R)TLSNYA (SEQ ID NO: 119);k). GRSFGAQGMEG (SEQ ID NO: 72);l). GFTLDLGAYA (SEQ ID NO: 81);m). GFTFGALA (SEQ ID NO: 86);n). GFTLS(G / S)YA (SEQ ID NO: 120);o). GSIFGGYN (SEQ ID NO: 2189);p). G(G / S)NFRILS (SEQ ID NO: 2209); andq). G(F / L)(A / T)F(R / S)(R / S)YA (SEQ ID NO: 2212).

[0345] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises a complementarity determining region 1 (CDR1) comprising an amino acid sequence selected from GRTFSSNL (SEQ ID NO: 1); GGTLANNG (SEQ ID NO: 6); GRTLDAYG (SEQ ID NO: 11); GRTLSDYA (SEQ ID NO: 23); GRTFSSYA (SEQ ID NO: 28); GRDFSNYV (SEQ ID NO: 33); GRSFSSYA (SEQ ID NO: 38); GRTFSTLA (SEQ ID NO: 43); GRTFSSDI (SEQ ID NO: 47); GRSFGDFA (SEQ ID NO: 52); GGTLSNYA (SEQ ID NO: 62); GRTLSNYA (SEQ ID NO: 67); GRSFGAQGMEG (SEQ ID NO: 72); GFTLDLGAYA (SEQ ID NO: 81); GFTFGALA (SEQ ID NO: 86); GFTLSGYA (SEQ ID NO: 95); GSIFGGYN (SEQ ID NO: 2189); GSNFRILS (SEQ ID NO: 2195); and GFTFSRYA (SEQ ID NO: 2201).

[0346] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) described herein may further comprise a complementarity determining region93323195900v2Attorney Docket No: 260525.0001282 (CDR2) comprising an amino acid sequence selected from (amino acids listed in a pair of brackets represent the possible amino acids at the particular position):a). VSWNGAST (SEQ ID NO: 2);b). DHR(S / T)GT (SEQ ID NO: 121);c). I(N / S)W(N / S / T)G(T / V)(D / G)T (SEQ ID NO: 122);d). LNW(N / S)G(D / E)ST (SEQ ID NO: 123);e). INWAD(E / T)T (SEQ ID NO: 124);f). INWSGG(S / T)T (SEQ ID NO: 125);g). ISWSDMSA (SEQ ID NO: 48);h). I(N / R)W(A / D / T)G(D / N)T(SEQ ID NO: 126);i). ISQ(S / T)S(D / S)T (SEQ ID NO: 127);j). (I / M)KWTGNT (SEQ ID NO: 128);k). ISN(S / T)GTTT (SEQ ID NO: 129);l). ISNDGEHI (SEQ ID NO: 87);m). ISWNGDIT (SEQ ID NO: 91);n). IT(G / S)(A / S)G(G / S)(N / S)T (SEQ ID NO: 130);o). IFISGN(D / N) (SEQ ID NO: 2207);p). (I / L)T(K / M / S)D(D / G)TT (SEQ ID NO: 2210); andq). ISS(A / G / S)(G / S)G(I / Y)(I / T / V) (SEQ ID NO: 2213).

[0347] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises a complementarity determining region 2 (CDR2) comprising an amino acid sequence selected from VSWNGAST (SEQ ID NO: 2); DHRSGT (SEQ ID NO: 7); ISWTGVDT (SEQ ID NO: 12); ISWTGTDT (SEQ ID NO: 19); ISWSGVDT (SEQ ID NO: 24); LNWSGEST (SEQ ID NO: 29); INWADET (SEQ ID NO: 34); INWSGGST (SEQ ID NO: 39); ISWSDMSA (SEQ ID NO: 48); IRWTGDT (SEQ ID NO: 53); ISQTSST (SEQ ID NO: 63); ISQSSDT(SEQ ID NO: 68); MKWTGNT (SEQ ID NO: 73); IKWTGNT (SEQ ID NO: 77); ISNTGTTT (SEQ ID NO: 82); ISNDGEHI (SEQ ID NO: 87); ISWNGDIT (SEQ ID NO: 91); ITSAGGST (SEQ ID NO: 96); IFISGNN (SEQ ID NO: 2190); ITSDDTT (SEQ ID NO: 2196); and ISSAGGYI (SEQ ID NO: 2202).

[0348] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from (amino acids listed in a pair of brackets represent the possible amino acids at the particular position):94323195900v2Attorney Docket No: 260525.000128a). (A / V)ASRL(P / V)FNSRSA(I / V)YTDRIYDS (SEQ ID NO: 100);b). AVRRSAWY(S / T)DSIYTVSQYDY (SEQ ID NO: 101);c). NAARSYSR(D / G / N)(G / Y)(E / R)PL(E / K)P(A / D)Y (SEQ ID NO: 102);d). AAASSWSRGG(A / G / I / V)PYGMDY (SEQ ID NO: 103);e). AADS(H / R)FRR(P / Y)(A / T / V)PG(I / Q)QYEY (SEQ ID NO: 104);f). NAA(K / R)SYHRDY(K / S)PL(K / S)(G / P)DY (SEQ ID NO: 105);g). AAAPSFGM(M / R / T)(I / N)PESYVHS (SEQ ID NO: 106);h). AANRGIMSMRLSRYDD (SEQ ID NO: 49);i). T(A / V)GP(A / T)MSYSRGGEF (SEQ ID NO: 107);j). (A / V)ADRGAISRSGAGM(D / N)Y (SEQ ID NO: 108);k). TAGP(A / S)IS(L / Y)SRGGEY (SEQ ID NO: 109);l). A(A / T)NGWGLDP(S / T)TYH(Y / D) (SEQ ID NO: 110);m). SAGWTRRIFQY (SEQ ID NO: 88);n). TAGQSISLSQGGE(H / Y) (SEQ ID NO: 111);o). KAGIRGE(T / V)Y (SEQ ID NO: 112);p). RAYNDGGEY (SEQ ID NO: 2191);q). HS(N / R)WYNL (SEQ ID NO: 2208); andr). (F / M / N)T(A / S)DY,wherein one or more non-alanine residues in the CDR3 sequence is optionally replaced with an alanine, and / or one or more alanine residues in the CDR3 sequence is optionally replaced with a glycine.

[0349] In some embodiments, one non-alanine residue in the CDR3 sequence is replaced with an alanine. In some embodiments, two non-alanine residues in the CDR3 sequence are replaced with an alanine. In some embodiments, three non-alanine residues in the CDR3 sequence are replaced with an alanine. In some embodiments, four non-alanine residues in the CDR3 sequence are replaced with an alanine. In some embodiments, five or more non-alanine residues in the CDR3 sequence are replaced with an alanine.

[0350] In some embodiments, one alanine residue in the CDR3 sequence is replaced with a glycine. In some embodiments, two alanine residues in the CDR3 sequence are replaced with a glycine. In some embodiments, three alanine residues in the CDR3 sequence are replaced with a glycine. In some embodiments, four alanine residues in the CDR3 sequence are replaced with a glycine. In some embodiments, five alanine or more residues in the CDR3 sequence are replaced with a glycine.95323195900v2Attorney Docket No: 260525.000128

[0351] In some embodiments, any of the above-described non-alanine residue replacement(s) with an alanine in the CDR3 sequence can be combined with any of the above-described alanine residue replacement(s) with a glycine in the CDR3 sequence.

[0352] In some embodiments, the CDR3 of the anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises an amino acid sequence selected froms). (A / G)(A / G)(A / G)(A / S)(A / S)(A / W)(A / S)(A / R)(A / G)(A / G)(A / G / l / V)(A / P)(A / Y)(A / G)(A / M)(A / D) (A / Y);t). (A / T)(A / G / V)(A / G)(A / P)(A / T)(A / M)(A / S)(A / Y)(A / S)(A / R)(A / G)(A / G)(A / E)(A / F);u). (A / R)(A / G)(A / Y)(A / N)(A / D)(A / G)(A / G)(A / E)(A / Y);v). (A / H)(A / S)(A / N / R)(A / W)(A / Y)(A / N)(A / L); andw). (A / F / M / N)(A / T)(A / G / S)(A / D)Y.

[0353] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from VASRLPFNSRSAIYTDRIYDS (SEQ ID NO: 3); AVRRSAWYSDSIYTVSQYDY (SEQ ID NO: 8); NAARSYSRDYEPLKPDY (SEQ ID NO: 13); NAARSYSRNYEPLKPDY (SEQ ID NO: 16); NAARSYSRGGEPLKPDY (SEQ ID NO: 20); NAARSYSRGGRPLEPAY (SEQ ID NO: 25);AAASSWSRGGVPYGMDY (SEQ ID NO: 30); AADSHFRRYTPGQQYEY (SEQ ID NO: 35); NAAKSYHRDYSPLSPDY (SEQ ID NO: 40); AAAPSFGMRNPESYVHS (SEQ ID NO: 44); AANRGIMSMRLSRYDD (SEQ ID NO: 49);TAGPTMSYSRGGEF (SEQ ID NO: 54); VADRGAISRSGAGMDY (SEQ ID NO: 64); AADRGAISRSGAGMDY (SEQ ID NO: 69); TAGPAISLSRGGEY (SEQ ID NO: 74); TAGPSISYSRGGEY (SEQ ID NO: 78); AANGWGLDPTTYHY (SEQ ID NO: 83); SAGWTRRIFQY (SEQ ID NO: 88); TAGQSISLSQGGEY (SEQ ID NO: 92); KAGIRGEVY (SEQ ID NO: 97); RAYNDGGEY (SEQ ID NO: 2191); HSRWYNL (SEQ ID NO: 2197); FTADY (SEQ ID NO: 2203);GAASSWSRGGVPYGMDY (SEQ ID NO: 2298); AGASSWSRGGVPYGMDY (SEQ ID NO: 2299);AAGSSWSRGGVPYGMDY (SEQ ID NO: 2300); AAAASWSRGGVPYGMDY (SEQ ID NO: 2301);AAASAWSRGGVPYGMDY (SEQ ID NO: 2302); AAASSASRGGVPYGMDY (SEQ ID NO: 2303);AAASSWARGGVPYGMDY (SEQ ID NO: 2304); AAASSWSAGGVPYGMDY (SEQ ID NO: 2305);AAASSWSRAGVPYGMDY (SEQ ID NO: 2306); AAASSWSRGAVPYGMDY (SEQ ID NO: 2307);AAASSWSRGGAPYGMDY (SEQ ID NO: 2308); AAASSWSRGGVAYGMDY (SEQ ID NO: 2309);AAASSWSRGGVPAGMDY (SEQ ID NO: 2310); AAASSWSRGGVPYAMDY (SEQ ID NO: 2311);AAASSWSRGGVPYGADY (SEQ ID NO: 2312); AAASSWSRGGVPYGMAY (SEQ ID NO: 2313);AAASSWSRGGVPYGMDA (SEQ ID NO: 2314); AAGPTMSYSRGGEF (SEQ ID NO: 2315); TGGPTMSYSRGGEF96323195900v2Attorney Docket No: 260525.000128(SEQ ID NO: 2316); TAAPTMSYSRGGEF (SEQ ID NO: 2317); TAGATMSYSRGGEF (SEQ ID NO: 2318);TAGPAMSYSRGGEF (SEQ ID NO: 2319); TAGPTASYSRGGEF (SEQ ID NO: 2320); TAGPTMAYSRGGEF (SEQ ID NO: 2321); TAGPTMSASRGGEF (SEQ ID NO: 2322); TAGPTMSYARGGEF (SEQ ID NO: 2323);TAGPTMSYSAGGEF (SEQ ID NO: 2324); TAGPTMSYSRAGEF (SEQ ID NO: 2325); TAGPTMSYSRGAEF (SEQ ID NO: 2326); TAGPTMSYSRGGAF (SEQ ID NO: 2327); TAGPTMSYSRGGEA (SEQ ID NO: 2328); AAYNDGGEY (SEQ ID NO: 2329); RGYNDGGEY (SEQ ID NO: 2330); RAANDGGEY (SEQ ID NO: 2331); RAYADGGEY (SEQ ID NO: 2332); RAYNAGGEY (SEQ ID NO: 2333); RAYNDAGEY (SEQ ID NO: 2334); RAYNDGAEY (SEQ ID NO: 2335); RAYNDGGAY (SEQ ID NO: 2336); RAYNDGGEA (SEQ ID NO: 2337); ASRWYNL (SEQ ID NO: 2338); HARWYNL (SEQ ID NO: 2339); HSAWYNL (SEQ ID NO: 2340); HSRAYNL (SEQ ID NO: 2341); HSRWANL (SEQ ID NO: 2342); HSRWYAL (SEQ ID NO: 2343); HSRWYNA (SEQ ID NO: 2344); ATADY (SEQ ID NO: 2345); FAADY (SEQ ID NO: 2346); FTGDY (SEQ ID NO: 2347); and FTAAY (SEQ ID NO: 2348).

[0354] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprisesi). a CDR1 comprising an amino acid sequence of GRTFSSNL (SEQ ID NO: 1), a CDR2 comprising an amino acid sequence of VSWNGAST (SEQ ID NO: 2), and a CDR3 comprising an amino acid sequence of (A / V)ASRL(P / V)FNSRSA(I / V)YTDRIYDS (SEQ ID NO: 100);ii). a CDR1 comprising an amino acid sequence of GGT(F / L)(A / S)N(D / N)G (SEQ ID NO: 113), a CDR2 comprising an amino acid sequence of DHR(S / T)GT (SEQ ID NO: 121), a CDR3 comprising an amino acid sequence of AVRRSAWY(S / T)DSIYTVSQYDY (SEQ ID NO: 101);iii). a CDR1 comprising an amino acid sequence of GRTL(D / N / S)(A / D / E)Y(A / G) (SEQ ID NO: 114), a CDR2 comprising an amino acid sequence of I(N / S)W(N / S / T)G(T / V)(D / G)T (SEQ ID NO: 122), a CDR3 comprising an amino acid sequence of NAARSYSR(D / G / N)(G / Y)(E / R)PL(E / K)P(A / D)Y (SEQ ID NO: 102);iv). a CDR1 comprising an amino acid sequence of GRTFS(N / S)YA (SEQ ID NO: 115), a CDR2 comprising an amino acid sequence of LNW(N / S)G(D / E)ST (SEQ ID NO: 123), a CDR3 comprising an amino acid sequence of AAASSWSRGG(A / G / I / V)PYGMDY (SEQ ID NO: 103);v). a CDR1 comprising an amino acid sequence of GRDFSNYV (SEQ ID NO: 33), a CDR2 comprising an amino acid sequence of INWAD(E / T)T (SEQ ID NO: 124), a CDR3 comprising an amino acid sequence of AADS(H / R)FRR(P / Y)(A / T / V)PG(I / Q)QYEY (SEQ ID NO: 104);vi). a CDR1 comprising an amino acid sequence of G( L / R)( l / S) FS( D / S)YA (SEQ ID NO: 116), a CDR2 comprising an amino acid sequence of INWSGG(S / T)T (SEQ ID NO: 125), a CDR3 comprising an amino acid sequence of NAA(K / R)SYHRDY(K / S)PL(K / S)(G / P)DY (SEQ ID NO: 105);97323195900v2Attorney Docket No: 260525.000128vii). a CDR1 comprising an amino acid sequence of GR(A / T)FSTLA (SEQ ID NO: 117), a CDR2 comprising an amino acid sequence of INWSGG(S / T)T (SEQ ID NO: 125), a CDR3 comprising an amino acid sequence of AAAPSFGM(M / R / T)(I / N)PESYVHS (SEQ ID NO: 106);viii). a CDR1 comprising an amino acid sequence of GRTFSSDI (SEQ ID NO: 47), a CDR2 comprising an amino acid sequence of ISWSDMSA (SEQ ID NO: 48), a CDR3 comprising an amino acid sequence of AANRGIMSMRLSRYDD (SEQ ID NO: 49);ix). a CDR1 comprising an amino acid sequence of GRSFGD(D / F / Y)A (SEQ ID NO: 118), a CDR2 comprising an amino acid sequence of I(N / R)W(A / D / T)G(D / N)T (SEQ ID NO: 126), a CDR3 comprising an amino acid sequence of T(A / V)GP(A / T)MSYSRGGEF (SEQ ID NO: 107);x). a CDR1 comprising an amino acid sequence of G(G / R)TLSNYA (SEQ ID NO: 119), a CDR2 comprising an amino acid sequence of ISQ(S / T)S(D / S)T (SEQ ID NO: 127), a CDR3 comprising an amino acid sequence of (A / V)ADRGAISRSGAGM(D / N)Y (SEQ ID NO: 108);xi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of (I / M)KWTGNT (SEQ ID NO: 128), a CDR3 comprising an amino acid sequence of TAGP(A / S)IS(L / Y)SRGGEY (SEQ ID NO: 109);xii). a CDR1 comprising an amino acid sequence of GFTLDLGAYA (SEQ ID NO: 81), a CDR2 comprising an amino acid sequence of ISN(S / T)GTTT (SEQ ID NO: 129), a CDR3 comprising an amino acid sequence of A(A / T)NGWGLDP(S / T)TYH(Y / D) (SEQ ID NO: 110);xiii). a CDR1 comprising an amino acid sequence of GFTFGALA (SEQ ID NO: 86), a CDR2 comprising an amino acid sequence of ISNDGEHI (SEQ ID NO: 87), a CDR3 comprising an amino acid sequence of SAGWTRRIFQY (SEQ ID NO: 88);xiv). a CDR1 comprising an amino acid sequence of GRTFS(N / S)YA (SEQ ID NO: 115), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), a CDR3 comprising an amino acid sequence of TAGQSISLSQGGE(H / Y) (SEQ ID NO: 111);xv). a CDR1 comprising an amino acid sequence of GFTLS(G / S)YA (SEQ ID NO: 120), a CDR2 comprising an amino acid sequence of IT(G / S)(A / S)G(G / S)(N / S)T (SEQ ID NO: 130), a CDR3 comprising an amino acid sequence of KAGIRGE(T / V)Y (SEQ ID NO: 112);xvi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGN(D / N) (SEQ ID NO: 2207), a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);98323195900v2Attorney Docket No: 260525.000128xvii). a CDR1 comprising an amino acid sequence of G(G / S)NFRILS (SEQ ID NO: 2209), a CDR2 comprising an amino acid sequence of (l / L)T(K / M / S)D(D / G)TT (SEQ ID NO: 2210), a CDR3 comprising an amino acid sequence of HS(N / R)WYNL (SEQ ID NO: 2208);xviii). a CDR1 comprising an amino acid sequence of G(F / L)(A / T)F(R / S)(R / S)YA (SEQ ID NO: 2212), a CDR2 comprising an amino acid sequence of ISS(A / G / S)(G / S)G( l / Y)( l / T / V) (SEQ ID NO: 2213), a CDR3 comprising an amino acid sequence of (F / M / N)T(A / S)DY;xix). a CDR1 comprising an amino acid sequence of GRTFS(N / S)YA (SEQ ID NO: 115), a CDR2 comprising an amino acid sequence of LNW(N / S)G(D / E)ST (SEQ ID NO: 123), a CDR3 comprising an amino acid sequence of (A / G)(A / G)(A / G)(A / S)(A / S)(A / W)(A / S)(A / R)(A / G)(A / G)(A / G / I / V)(A / P)(A / Y)(A / G)(A / M)(A / D)(A / Y);xx). a CDR1 comprising an amino acid sequence of GRSFGD(D / F / Y)A (SEQ ID NO: 118), a CDR2 comprising an amino acid sequence of I(N / R)W(A / D / T)G(D / N)T (SEQ ID NO: 126), a CDR3 comprising an amino acid sequence of (A / T)(A / G / V)(A / G)(A / P)(A / T)(A / M)(A / S)(A / Y)(A / S)(A / R)(A / G)(A / G)(A / E)(A / F); xxi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGN(D / N) (SEQ ID NO: 2207), a CDR3 comprising an amino acid sequence of (A / R)(A / G)(A / Y)(A / N)(A / D)(A / G)(A / G)(A / E)(A / Y);xxii). a CDR1 comprising an amino acid sequence of G(G / S)NFRILS (SEQ ID NO: 2209), a CDR2 comprising an amino acid sequence of (l / L)T(K / M / S)D(D / G)TT (SEQ ID NO: 2210), a CDR3 comprising an amino acid sequence of (A / H)(A / S)(A / N / R)(A / W)(A / Y)(A / N)(A / L); orxxiii). a CDR1 comprising an amino acid sequence of G(F / L)(A / T)F(R / S)(R / S)YA (SEQ ID NO: 2212), a CDR2 comprising an amino acid sequence of ISS( A / G / S)(G / S)G( l / Y)( l / T / V) (SEQ ID NO: 2213), a CDR3 comprising an amino acid sequence of (A / F / M / N)(A / T)(A / G / S)(A / D)Y.

[0355] Provided herein are anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) comprising a CDR1 (CDR1) comprising an amino acid sequence selected from any of the CDR1 amino acid sequences listed in Table 1-3 or Table 6, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0356] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as singledomain antibody) comprises a CDR1 comprising an amino acid sequence selected from SEQ ID NOs: 1, 6, 11, 23, 28, 33, 38, 43, 47, 52, 62, 67, 72, 81, 86, 95, 762-1084, 2189, 2195, 2201, and 2252-2261, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.99323195900v2Attorney Docket No: 260525.000128

[0357] Provided herein are anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) comprising a CDR2 (CDR2) comprising an amino acid sequence selected from any of the CDR2 amino acid sequences listed in Table 1-3 or Table 6, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0358] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as singledomain antibody) comprises a CDR2 comprising an amino acid sequence selected from SEQ ID NOs: 2, 7, 12, 19, 24, 29, 34, 39, 48, 53, 63, 68, 73, 77, 82, 87, 91, 96, 1085-1407, 2190, 2196, 2202, and 2262-2271, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0359] Provided herein are anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) comprising a CDR3 (CDR3) comprising an amino acid sequence selected from any of the CDR3 amino acid sequences listed in Table 1-3 or Table 6, or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0360] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as singledomain antibody) comprises a CDR3 comprising an amino acid sequence selected from SEQ ID NOs: 3, 8, 13, 16, 20, 25, 30, 35, 40, 44, 49, 54, 64, 69, 74, 78, 83, 88, 92, 97, 1408-1730, 2191, 2197, 2203, 2272-2277, and 2298-2348 or a similar sequence thereof having at least 70%, at least 80%, at least 90%, or at least 95% sequence identity.

[0361] Provided herein are anti-TRAILR2 antigen-binding domains (e.g., antibodies such as singledomain antibodies) comprising a set of three CDRs ( / . e., CDR1-CDR2-CDR3) contained within any of the exemplary anti-TRAILR2 VHH antibodies listed in Table 1-3, Table 1-4, or Table 6. In certain embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprisesi) a CDR1 comprising an amino acid sequence of GRTFSSNL (SEQ ID NO: 1), a CDR2 comprising an amino acid sequence of VSWNGAST (SEQ ID NO: 2), and a CDR3 comprising an amino acid sequence of VASRLPFNSRSAIYTDRIYDS (SEQ ID NO: 3);ii) a CDR1 comprising an amino acid sequence of GGTLANNG (SEQ ID NO: 6), a CDR2 comprising an amino acid sequence of DHRSGT (SEQ ID NO: 7), a CDR3 comprising an amino acid sequence of AVRRSAWYSDSIYTVSQYDY (SEQ ID NO: 8);iii) a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), a CDR3 comprising an amino acid sequence of NAARSYSRDYEPLKPDY (SEQ ID NO: 13);100323195900v2Attorney Docket No: 260525.000128iv) a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), a CDR3 comprising an amino acid sequence of NAARSYSRNYEPLKPDY (SEQ ID NO: 16);v) a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGTDT (SEQ ID NO: 19), a CDR3 comprising an amino acid sequence of NAARSYSRGGEPLKPDY (SEQ ID NO: 20);vi) a CDR1 comprising an amino acid sequence of GRTLSDYA (SEQ ID NO: 23), a CDR2 comprising an amino acid sequence of ISWSGVDT (SEQ ID NO: 24), a CDR3 comprising an amino acid sequence of NAARSYSRGGRPLEPAY (SEQ ID NO: 25);vii) a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);viii) a CDR1 comprising an amino acid sequence of GRDFSNYV (SEQ ID NO: 33), a CDR2 comprising an amino acid sequence of INWADET (SEQ ID NO: 34), a CDR3 comprising an amino acid sequence of AADSHFRRYTPGQQYEY (SEQ ID NO: 35);ix) a CDR1 comprising an amino acid sequence of GRSFSSYA (SEQ ID NO: 38), a CDR2 comprising an amino acid sequence of INWSGGST (SEQ ID NO: 39), a CDR3 comprising an amino acid sequence of NAAKSYHRDYSPLSPDY (SEQ ID NO: 40);x) a CDR1 comprising an amino acid sequence of GRTFSTLA (SEQ ID NO: 43), a CDR2 comprising an amino acid sequence of INWSGGST (SEQ ID NO: 39), a CDR3 comprising an amino acid sequence of AAAPSFGMRNPESYVHS (SEQ ID NO: 44);xi) a CDR1 comprising an amino acid sequence of GRTFSSDI (SEQ ID NO: 47), a CDR2 comprising an amino acid sequence of ISWSDMSA (SEQ ID NO: 48), a CDR3 comprising an amino acid sequence of AANRGIMSMRLSRYDD (SEQ ID NO: 49);xii) a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);xiii) a CDR1 comprising an amino acid sequence of GGTLSNYA (SEQ ID NO: 62), a CDR2 comprising an amino acid sequence of ISQTSST (SEQ ID NO: 63), a CDR3 comprising an amino acid sequence of VADRGAISRSGAGMDY (SEQ ID NO: 64);101323195900v2Attorney Docket No: 260525.000128xiv) a CDR1 comprising an amino acid sequence of GRTLSNYA (SEQ ID NO: 67), a CDR2 comprising an amino acid sequence of ISQSSDT (SEQ ID NO: 68), a CDR3 comprising an amino acid sequence of AADRGAISRSGAGMDY (SEQ ID NO: 69);xv) a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of MKWTGNT (SEQ ID NO: 73), a CDR3 comprising an amino acid sequence of TAGPAISLSRGGEY (SEQ ID NO: 74);xvi) a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78);xvii) a CDR1 comprising an amino acid sequence of GFTLDLGAYA (SEQ ID NO: 81), a CDR2 comprising an amino acid sequence of ISNTGTTT (SEQ ID NO: 82), a CDR3 comprising an amino acid sequence of AANGWGLDPTTYHY (SEQ ID NO: 83);xviii) a CDR1 comprising an amino acid sequence of GFTFGALA (SEQ ID NO: 86), a CDR2 comprising an amino acid sequence of ISNDGEHI (SEQ ID NO: 87), a CDR3 comprising an amino acid sequence of SAGWTRRIFQY (SEQ ID NO: 88);xix) a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92);xx) a CDR1 comprising an amino acid sequence of GFTLSGYA (SEQ ID NO: 95), a CDR2 comprising an amino acid sequence of ITSAGGST (SEQ ID NO: 96), a CDR3 comprising an amino acid sequence of KAGIRGEVY (SEQ ID NO: 97);xxi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);xxii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);xxiii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);102323195900v2Attorney Docket No: 260525.000128xxiv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of GAASSWSRGGVPYGMDY (SEQ ID NO: 2298);xxv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AGASSWSRGGVPYGMDY (SEQ ID NO: 2299);xxvi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAGSSWSRGGVPYGMDY (SEQ ID NO: 2300);xxvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAAASWSRGGVPYGMDY (SEQ ID NO: 2301);xxviii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASAWSRGGVPYGMDY (SEQ ID NO: 2302);xxix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSASRGGVPYGMDY (SEQ ID NO: 2303);xxx). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWARGGVPYGMDY (SEQ ID NO: 2304);xxxi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSAGGVPYGMDY (SEQ ID NO: 2305);xxxii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRAGVPYGMDY (SEQ ID NO: 2306);xxxiii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGAVPYGMDY (SEQ ID NO: 2307);103323195900v2Attorney Docket No: 260525.000128xxxiv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGAPYGMDY (SEQ ID NO: 2308);xxxv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVAYGMDY (SEQ ID NO: 2309);xxxvi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPAGMDY (SEQ ID NO: 2310);xxxvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYAMDY (SEQ ID NO: 2311);xxxviii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGADY (SEQ ID NO: 2312);xxxix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMAY (SEQ ID NO: 2313);xl). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDA (SEQ ID NO: 2314);xli). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of AAGPTMSYSRGGEF (SEQ ID NO: 2315);xlii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TGGPTMSYSRGGEF (SEQ ID NO: 2316);xliii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAAPTMSYSRGGEF (SEQ ID NO: 2317);104323195900v2Attorney Docket No: 260525.000128xliv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGATMSYSRGGEF (SEQ ID NO: 2318);xlv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPAMSYSRGGEF (SEQ ID NO: 2319);xlvi). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTASYSRGGEF (SEQ ID NO: 2320);xlvii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMAYSRGGEF (SEQ ID NO: 2321);xlviii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSASRGGEF (SEQ ID NO: 2322);xlix). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYARGGEF (SEQ ID NO: 2323);I), a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSAGGEF (SEQ ID NO: 2324);li). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRAGEF (SEQ ID NO: 2325);lii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGAEF (SEQ ID NO: 2326);liii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGAF (SEQ ID NO: 2327);105323195900v2Attorney Docket No: 260525.000128liv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEA (SEQ ID NO: 2328);Iv). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of AAYNDGGEY (SEQ ID NO: 2329);Ivi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RGYNDGGEY (SEQ ID NO: 2330);Ivii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAANDGGEY (SEQ ID NO: 2331);Iviii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYADGGEY (SEQ ID NO: 2332);lix). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNAGGEY (SEQ ID NO: 2333);lx), a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDAGEY (SEQ ID NO: 2334);Ixi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGAEY (SEQ ID NO: 2335);Ixii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGAY (SEQ ID NO: 2336);Ixiii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEA (SEQ ID NO: 2337);106323195900v2Attorney Docket No: 260525.000128Ixiv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of ASRWYNL (SEQ ID NO: 2338);Ixv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HARWYNL (SEQ ID NO: 2339);Ixvi). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSAWYNL (SEQ ID NO: 2340);Ixvii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRAYNL (SEQ ID NO: 2341);Ixviii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWANL (SEQ ID NO: 2342);Ixix). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYAL (SEQ ID NO: 2343);Ixx). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNA (SEQ ID NO: 2344);Ixxi). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of ATADY (SEQ ID NO: 2345);Ixxii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FAADY (SEQ ID NO: 2346);Ixxiii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTGDY (SEQ ID NO: 2347); or107323195900v2Attorney Docket No: 260525.000128Ixxiv). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTAAY (SEQ ID NO: 2348).

[0362] In certain embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprisesi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);ii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);iii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);iv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197); orv). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203),wherein one or more non-alanine residues in the CDR3 sequence is optionally replaced with an alanine, and / or one or more alanine residues in the CDR3 sequence is optionally replaced with a glycine.

[0363] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure comprises:i). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);ii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);108323195900v2Attorney Docket No: 260525.000128iii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);iv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);v). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203); orvi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92).

[0364] In some embodiments, the one or more anti-TRAILR2 antigen-binding domains comprise: i). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);ii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);iii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);iv). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92).109323195900v2Attorney Docket No: 260525.000128

[0365] In a related embodiment, provided herein are anti-TRAILR2 antigen-binding domains (e.g., antibodies such as single-domain antibodies) comprising a set of three CDRs ( / .e., CDR1-CDR2-CDR3) contained within a VHH amino acid sequence as defined by any of the exemplary anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6. For example, provided herein are antibodies, or antigen-binding fragments thereof, comprising the set of CDR1-CDR2-CDR3 amino acid sequences contained within a VHH amino acid sequence selected from SEQ ID NOs: 4, 5, 9, 10, 14, 15, 17, 18, 21, 22, 26, 27, 31, 32, 36, 37, 41, 42, 45, 46, 50, 51, 55, 56, 65, 66, 70, 71, 75, 76, 79, 80, 84, 85, 89, 90, 93, 94, 98, 99, 146-468, 469-761, 2192, 2194, 2198, 2200, 2204, 2206, 2214-2233, 2234-2251, 2354-2404, 4662-4677, and 4692-4697.

[0366] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can includea) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 4;b) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 9;c) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 14;d) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 17;e) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 21;f) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 26;g) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 31;h) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 36;i) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 41;j) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 45;110323195900v2Attorney Docket No: 260525.000128k) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 50;l) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 55;m) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 65;n) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 70;o) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 75;p) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 79;q) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 84;r) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 89;s) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 93;t) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 98;u) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 2192;v) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 2198; orw) a variable domain that comprises a CDR1, CDR2, and CDR3 contained within a VHH comprising the amino acid sequence of SEQ ID NO: 2204.

[0367] In an embodiment provided herein, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a VHH amino acid sequence selected from SEQ ID NOs: 4, 9, 14, 17, 21, 26, 31, 36, 41, 45, 50, 55, 65, 70, 75, 79, 84, 89, 93, 98, 146-468, 2192, 2198, 2204, and 2214-2233, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%sequence identity.111323195900v2Attorney Docket No: 260525.000128

[0368] In an embodiment provided herein, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a VHH amino acid sequence selected from SEQ ID NOs: 4, 9, 14, 17, 21, 26, 31, 36, 41, 45, 50, 55, 65, 70, 75, 79, 84, 89, 93, 98, 2192, 2198, and 2204, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0369] In an embodiment provided herein, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a VHH amino acid sequence selected from SEQ ID NOs: 14, 31, 55, 2192, 2198, and 2204, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0370] In some embodiments of any of the above-described antigen-binding proteins, the VHH may be a humanized VHH.

[0371] In an embodiment provided herein, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a humanized VHH amino acid sequence selected from SEQ ID NOs: 5, 10, 15, 18, 22, 27, 32, 37, 42, 46, 51, 56, 66, 71, 76, 80, 85, 90, 94, 99, 469-761, 2194, 2200, 2206, 2234-2251, 2354-2404, 4662-4677, and 4692-4697, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0372] In an embodiment provided herein, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a humanized VHH amino acid sequence selected from SEQ ID NOs: 5, 10, 15, 18, 22, 27, 32, 37, 42, 46, 51, 56, 66, 71, 76, 80, 85, 90, 94, 99, 2194, 2200, 2206, 2354-2404, 4662-4677, and 4692-4697, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0373] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a humanized VHH amino acid sequence selected from any one of SEQ ID NOs: 32, 56, 76, 80, 94, 99, 2194, 2200, 2206, 2250, 2379, 2381, and 4662-4677, or an amino acid sequence having at least 75% identity thereto.112323195900v2Attorney Docket No: 260525.000128

[0374] In some embodiments, an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) of the present disclosure can include a humanized VHH amino acid sequence selected from any one of SEQ ID NOs: 4662-4677 and 4692-4697, or a similar sequence thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity.

[0375] In some embodiments, a multispecific antigen-binding protein of the present disclosure also comprises an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) that competes for binding to TRAILR2 with any one of the exemplary anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6.

[0376] In some embodiments, a multispecific antigen-binding protein of the present disclosure also comprises an anti-TRAILR2 antigen-binding domain (e.g., antibody such as a single-domain antibody) that binds to the same epitope on TRAILR2 as any one of the exemplary anti-TRAILR2 VHH antibodies listed in Table 1-1, Table 1-2, or Table 6.Anti-CDHl 7 Antigen-binding domains

[0377] The present disclosure provides antigen-binding domains (e.g., antibodies, such as singledomain antibodies) that bind to cadherin 17 (CDH17).

[0378] Cadherin 17 (CDH17) is a calcium-dependent cell adhesion glycoprotein belonging to the 7D cadherin subfamily within the cadherin superfamily. It consists of seven extracellular cadherin (EC) repeats with a total length of 764 amino acids, a single transmembrane helix and a very short intracellular domain of only 23 amino acids, lacking binding sites for interactions with the cytoskeleton proteins such as catenins. CDH17 is predicted to harbor eight N-glycans located throughout all ECs except for ECI, with several of them confirmed experimentally. Between the linkers of each EC domain (except for EC2 and EC3) CDH17 exhibits the typical canonical calcium binding motif that coordinates three calcium ions, which were shown to contribute to the rigidity of the protein.

[0379] Similar to other cadherins, CDH17 contributes to cell adhesion via trans-interaction with cadherins of neighboring cells. However, instead of dimerizing via ECI and EC2, it was shown that in CDH17 EC2 of one chain interacts with EC4 of a second chain. Further it was suggested that additional EC domains may be involved in the dimerization.

[0380] In addition, it was reported that CDH17 comprises an RGD (Arg-Gly-Asp) tripeptide motif in EC6, through which the protein interacts with integrin a2pl. This CDH17 a201 interaction was associated with aggressive forms of cancer during late-stage metastasis. The soluble form of CDH17 was113323195900v2Attorney Docket No: 260525.000128reported to contain the RGD motif suggesting proteolytic cleavage C-terminal of EC6 or even more proximal to the membrane.

[0381] In some embodiments, anti-CDH17 antigen-binding domains that specifically bind to cadherin 17 (CDH17) described herein can act as an anchor and / or a guide to a target cancer cell, while avoiding impact on CDH17 biology. Since CDH17 is not expressed on normal adult hepatocytes, CDH17 can be used as a targeting antigen so as to avoid liver toxicity that has plagued various therapeutic molecules aimed at targeting cancer. An anti-CDH17 antigen-binding domain of the present disclosure can target membrane proximal epitopes of CDH17 in order to not impair its dimerization. In some embodiments, membrane proximal targeting of can avoid binding to potentially shed CDH17 thus retaining targeting to tumor cells. In some embodiments, binding of the anti-CDH17 antigen-binding domains of the present disclosure to the integrin binding tripeptide RGD motif in extracellular cadherin domain 6 (EC6) of human CDH17 can be avoided.

[0382] In various embodiment, anti-CDH17 antigen-binding domains described herein can specifically bind to CDH17 from human, cyno and / or mouse.

[0383] In some embodiments, a human CDH17 is encoded by a CDH17 gene (NCBI Gene ID 1015; AA Sequence: >sp | Q128641 CAD17_HUMAN Cadherin-17 0S=Homo sapiens OX=9606 GN=CDH17 PE=1 S\Z=3 ) and has the amino acid sequence MILQAHLHSLCLLMLYLATGYGQEGKFSGPLKPMTFSIYEGQEPSQIIFQFKANPPAVTFELTGETDNIFVIEREGLLYYNR ALDRETRSTHNLQVAALDANGIIVEGPVPITIKVKDINDNRPTFLQSKYEGSVRQNSRPGKPFLYVNATDLDDPATPNGQ LYYQIVIQLPMINNVMYFQINNKTGAISLTREGSQELNPAKNPSYNLVISVKDMGGQSENSFSDTTSVDIIVTENIWKAP KPVEMVENSTDPHPIKITQVRWNDPGAQYSLVDKEKLPRFPFSIDQEGDIYVTQPLDREEKDAYVFYAVAKDEYGKPLS YPLEIHVKVKDINDNPPTCPSPVTVFEVQENERLGNSIGTLTAHDRDEENTANSFLNYRIVEQ. TPKLPMDGLFLIQ. TYAG MLQLAKQSLKKQDTPQYNLTIEVSDKDFKTLCFVQINVIDINDQIPIFEKSDYGNLTLAEDTNIGSTILTIQATDADEPFTGS SKILYHIIKGDSEGRLGVDTDPHTNTGYVIIKKPLDFETAAVSNIVFKAENPEPLVFGVKYNASSFAKFTUVTDVNEAPQFS QHVFQAKVSEDVAIGTKVGNVTAKDPEGLDISYSLRGDTRGWLKIDHVTGEIFSVAPLDREAGSPYRVQVVATEVGGSS LSSVSEFHLILMDVNDNPPRLAKDYTGLFFCHPLSAPGSLIFEATDDDQHLFRGPHFTFSLGSGSLQNDWEVSKINGTHA RLSTRHTEFEEREYVVLIRINDGGRPPLEGIVSLPVTFCSCVEGSCFRPAGHQTGIPTVGMAVGILLTTLLVIGIILAVVFIRIK KDKGKDNVESAQASEVKPLRS (UniProtKB Accession No. Q12864) (SEQ. ID NO: 4104).

[0384] In some embodiments, a cyno CDH17 is encoded by a CDH17 gene (NCBI Gene ID 102129651; AA Sequence: >tr| AOA2K5X8I8| AOA2K5X8I8_MACFA Cadherin 17 OS=Macaca fascicularis OX=9541 GN=CDH17 PE=4 SV=2) and has the amino acid sequence MLHSYCANKLLEGTTLKVLRISYLVIEPVKILSEIGDLLQIPQLCFFPYSLICCFFPQFKANPPAVTFELTGETDNIFKIEQEGLL114323195900v2Attorney Docket No: 260525.000128YYTKALDRETRSTHNLQVAALDANGAIVEGPVPITIEVKDVNDNRPTFLQSKYEGSVRQNSRPGKPFLYVNATDLDDPA TPNGQLSYQIVIQLPMINNVMYFQINNKTGGISLTREGSQELNPAKNPSYNLVISVKDMGGQSENSFSDTTSVDIIVTEN IWKAPEPVEMVENSTDPHPIKITQVRWNDPGAQYSLVDKEKLPRFPFSIDQEGDIYVTQPLDREEKDAYVFYAVAKDEY GKPLSYPLEIHVKVQDINDNPPTCPSPVTVFEVQENERLGNSIGALTAHDSDEENTANSLLNYRIVEQTPKLPMDGLFLIQ TYAGMLQLAKQSLKKQDTPQYNLTIEVSDKDFKTLCFVQINVIDINDQIPIFEKSDYGNLTLAEDTNVGSTILTIQATDADE PFTGSSKILYHVIKGDSEGRLGVDTDPHTNTGYVIIKKPLDFETAAISNIVFKAENPEPLVFGVTYNASSFAKFTLFVTDVNE APEFSQYVFQAKVSEDVAIGTKVGNVTAKDPEGLDISYSLRGDTRGWLKIDHVTGEIFSVAPLDREAGSPYRVQVVATEV GGSSLSSVSQFHLILTDVNDNPPRLAKDYMDLYFCHPLSAPGSLIFEATDDDQHLFRGPHFTFSIASESLQNDWQVSKIN GTHARLSTRHTDFEEKEYVVSIRINDGGRPPLESTVSLTVTFCSCGEDGCFRPAGHQPGIPTVGMAVGILLTTLLVIGIILA VVFIRMKTDKGKDNVESAQASEVKPLRS (UniProtKB Accession No. A0A2K5X8I8) (SEQ ID NO: 4105).

[0385] In some embodiments, a mouse CDH17 is encoded by a Cdhl7 gene (NCBI Gene ID 12557; AA Sequence: >sp | Q9R1001 CAD17_MOUSE Cadherin-17 OS=Mus musculus OX=10090 GN=Cdhl7 PE=1 SV=1) and has the amino acid sequence MVSAQLHFLCLLTLYLTCGYGEEGKFSGPLKPMTFSIFEGQEPSQVIFQFKTNPPAVTFELTGETDGIFKIEKDGLLYHTRA LDRETRAVHHLQLAALDSHGAIVDGPVPITIEVKDINDNRPTFLQSKYEGSVRQNSRPGKPFMYVNATDLDDPATPNG QLFYQIVIQLPQINDVMYFQIDSKTGAISLTPEGSQELDPVKNPSYNLVVSVKDMGGQSENSFSDTTYVDISIRENIWKA PEPVEIRENSTDPHPIKITQVQWNDPGAQYSLVNKEKLSPFPFSIDQEGNIYVTQALDREEKNSHVFFATAKDENGKPLA YPLEIYVKVIDINDNPPTCLSPVTVFEVQENEPLGNSIGIFEAHDMDEANNINSILKYKLVDQTPKVPSDGLFLIGEYEGKV QLSKQSLKKQDSPQYNLSIEVSDVDFKTLCYIQVNVIDINDQIPIFETSNYGSKTLSEDTAIGSTILIIQATDADEPFTGSSKIL YKIVQGDTEGRLEWTDPTTNAGYVKIKKPLDFETQPVSSIVFQAENPEPLVKGIEYNASSFASFEUVTDVNEVPVFPQRI FQANVSEDAAVGSRVGNVTARDPEGLTVSYSLKGNMRGWLKIDSVTGEIFSAAPLDRETESVYRVQVVATEVGGSSLSS TADFHLVLTDVNDNPPRLAKDYTGLFFCHPLSAPGSLIFEVTDDDQQSLRRPKFTFALGREGLQSDWEVSKINGTHARLS TRHTRFEEQVYNIPIRINDGGQPPMEGTVFLPVTFCQCVEGSCFRPAGRQDGIPTVGMAVGILLTTFLVIGIILAVVFIRM RKDKVENPQSPENKPLRS (UniProtKB Accession No. Q9R100) (SEQ ID NO: 4106).

[0386] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure bind to human CDH17. In some embodiments, anti-CDH17 antigen-binding domains [e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to human CDH17 with a KDof less than about 1x10“® M, for example, less than about 5xl0"7M, less than about 3xl0“7M, less than about IxlO-7M, less than about 8xl0"8M, less than about 5xl0-8M, less than about 3xl0-8M, less than about IxlO-8M, less than about 8xl0-9M, less than about 5xio-9M, less than about 3xl0“9M, or less than about IxlO-9M, or about IxlO’10to IxlO-9M, IxlO’10to 5xl0-9M, about IxlO’10to IxlO-8M, about115323195900v2Attorney Docket No: 260525.000128lxlO-10to 5xl0"8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8M, about lxlO-9to lxlO-7M, or about lxlO-8to lxlO-7M.

[0387] In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human CDH17 with a KDof less than about 3xl0“7M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human CDH17 with a KDof less than about 2.2xl0“7M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human CDH17 with a KDof less than about lxlO-7M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human CDH17 with a KDof about 1x1010to 5xl0“8M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human CDH17 with a KDof less than about 160 nM, 150 nM, 140 nM, 130 nM, 120 nM, 110 nM, 100 nM, 90 nM, 80 nM, 70 nM, 60 nM, 50 nM, 40 nM, 30 nM, 25 nM, 20 nM, 15 nM (e.g., about 12 nM, 11 nM, 10 nM, 9nM, 8 nM, 7 nM, 6 nM, 5 nM, 4 nM, 3 nM, 2nM, or 1 nM, or less).

[0388] In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to human CDH17 with a KDof about 1.54 x 10'7M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to human CDH17 with a KDof about 1.37 x 10'7M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to human CDH17 with a KDof about 6.2 x 10'8M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to human CDH17 with a KDof about 3.2 x 10’8M.

[0389] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure bind to cynomolgus monkey ("cyno") CDH17. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to cyno CDH17 with a KDof less than about lxlO-5M, for example, less than about 5xl0-7M, less than about 3xl0-7M, less than about lxlO-7M, less than about 8xl0-8M, less than about 5xl0-8M, less than about 3xl0-8M, less than about 2xl0-8M, less than about lxlO-8M, less than about 8xl0-9M, less than about 5xl0-9M, less than about 3xl0-9M, or less than about lxlO-9M, or about lxlO-10to lxlO-9M, lxlO-10to 5xl0-9M, about lxlO-10to lxlO-8M, about lxlO-10to 5xl0-8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8116323195900v2Attorney Docket No: 260525.000128M, about 1x109to 1x107M, about 1x109to 2x107M, about 1x109to 5x107M, about 1x108to lxlO-7M, about lxlO-8to 2xl0-7M, about lxl0“8to 5xl0“7M, or about lxl0“8to lxl0“6M.

[0390] In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to cyno CDH17 with a KDof about 1.1 x IO-8M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to cyno CDH17 with a KDof about 1 x 10’8M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to cyno CDH17 with a KDof about 9.7 x 10'9M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as single-domain antibodies) of the present disclosure may bind to cyno CDH17 with a KDof about 5.8 x 109M.

[0391] In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to cyno CDH17 with a KDof less than about 3×10-7M. In some embodiments, anti-CDH17 antigen-binding domains (e.g., antibodies such as singledomain antibodies) of the present disclosure may bind to cyno CDH17 with a KD of about lxlO-9to lxlO-7M.

[0392] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure do not bind to cyno CDH17.

[0393] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure bind to mouse CDH17. In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure may bind to mouse CDH17 with a KDof less than about lxlO-6M, for example, less than about 5xl0-7M, less than about 3xl0-7M, less than about lxlO-7M, less than about 8xl0-8M, less than about 5xl0-8M, less than about 3xl0-8M, less than about lxlO-8M, less than about 8xl0-9M, less than about 5xl0-9M, less than about 3xl0-9M, or less than about lxlO-9M, or about lxlO-10to lxlO-9M, lxlO-10to 5xl0-9M, about lxlO-10to lxlO-8M, about lxlO-10to 5xl0-8M, about lxlO-9to lxlO-8M, about lxlO-9to 5xl0-8M, about lxlO-9to lxlO-7M, about lxlO-9to 2xl0-7M, about lxlO-9to 5xl0-7M, about lxlO-8to lxlO-7M, about lxlO-8to 2xl0-7M, about lxlO-8to 5xl0-7M, or about lxlO-8to lxlO-6M. In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure may bind to mouse CDH17 with a KDof less than about 2xl0-7M (e.g., about 160 nM, 120 nM, 115 nM, 110 nM, 100 nM, 95 nM, 90 nM, 75 nM, 50 nM, 45 nM, 30 nM, 20 nM, or less).

[0394] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure may bind to mouse CDH17 with a KDof about lxlO-6M. In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure may bind to mouse CDH17 with a KDof about 7xl0-8M. In some117323195900v2Attorney Docket No: 260525.000128embodiments, anti-CDH17 antigen-binding domains of the present disclosure may bind to mouse CDH17 with a KDof about 5.2xl0-8M.

[0395] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure do not bind to mouse CDH17.

[0396] In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure bind to rat CDH17. In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure may bind to rat CDH17 with a KDof less than about lxlO’6M, for example, less than about 5xl0“7M, less than about 3xl0“7M, less than about lxlO’7M, less than about 8xl0“8M, less than about 5xl0“8M, less than about 3xl0“8M, less than about lxlO’8M, less than about 8xl0’9M, less than about 5xl0“9M, less than about 3x109M, or less than about 1x109M, or about 1x1010to 1x109M, 1x1010to 5x109M, about lxlO’10to lxlO’8M, about lxlO’10to 5xl0’8M, about lxlO’9to lxlO’8M, about lxlO’9to 5xl0’8M, about lxlO’9to lxlO’7M, about lxlO"9to 2xl0"7M, about lxlO’9to 5xl0“7M, about lxlO’8to lxlO’7M, about lxlO’8to 2xl0’7M, about lxlO’8to 5xl0’7M, or about lxlO’8to lxlO’6M. In some embodiments, anti-CDH17 antigen-binding domains of the present disclosure do not bind to rat CDH17.

[0397] In some embodiments, anti-CDH17 antigen-binding domains described herein do not block dimerization of CDH17. In some embodiments, anti-CDH17 antigen-binding domains described herein do not impact CDH17 biological functions. In some embodiments, anti-CDH17 antigen-binding domains described herein do not bind to shed CDH17 thus retaining targeting to tumor cells.

[0398] In some embodiments, anti-CDH17 antigen-binding domains described herein do not bind to the integrin binding RGD (Arg-Gly-Asp) motif in extracellular cadherin domain 6 (EC6) of human CDH17.

[0399] In some embodiments, anti-CDH17 antigen-binding domains described herein bind to the extracellular cadherin domain 6 (EC6) and / or extracellular cadherin domain 7 (EC7) of human CDH17.

[0400] In some embodiments, anti-CDH17 antigen-binding domains described herein bind to an extracellular region C-terminal to the RGD motif in EC6 of human CDH17.

[0401] In some embodiments, anti-CDH17 antigen-binding domains described herein bind to the extracellular cadherin domain 1 (ECI) of human CDH17.

[0402] Binding affinity of a molecular interaction between two molecules can be measured via various techniques, such as surface plasmon resonance (SPR), bio-layer interferometry (BLI), enzyme-linked immunosorbent assay (ELISA), equilibrium dialysis, fluorescent-activated cell sorting (FACS), flow cytometry binding assays, or isothermal titration calorimetry (ITC), and the like. Surface plasmon resonance is a biosensor technique that allows for the analysis of real-time biospecific interactions by detection of alterations in protein concentrations within a biosensor matrix, where one molecule is118323195900v2Attorney Docket No: 260525.000128immobilized on the biosensor chip and the other molecule is passed over the immobilized molecule underflow conditions (see e.g., Ober etal. 2001, Intern. Immunology 13: 1551-1559). SPR can for example be performed using the BIACORE® system or Carterra LSA system. Another biosensor technique that can be used to determine affinities of biomolecular interactions is bio-layer interferometry (BLI) (see e.g., Abdiche et al. 2008, Anal. Biochem. 377: 209-217). Bio-layer Interferometry is a label-free optical technique that analyzes the interference pattern of light reflected from two surfaces: an internal reference layer (reference beam) and a layer of immobilized protein on the biosensor tip (signal beam). A change in the number of molecules bound to the tip of the biosensor causes a shift in the interference pattern, reported as a wavelength shift (nm), the magnitude of which is a direct measure of the number of molecules bound to the biosensor tip surface. Since the interactions can be measured in real-time, association and dissociation rates and affinities can be determined. BLI can for example be performed using the Octet® Systems. Alternatively, affinities can be measured in Kinetic Exclusion Assay (KinExA) (see e.g., Drake et al. 2004, Anal. Biochem., 328: 35-43), which is a solution-based method to measure true equilibrium binding affinity and kinetics of unmodified molecules. Equilibrated solutions of an antibody / antigen complex are passed over a column with beads precoated with antigen (or antibody), allowing the free antibody (or antigen) to bind to the coated molecule. Detection of the antibody (or antigen) thus captured is accomplished with a fluorescently labeled protein binding the antibody (or antigen).

[0403] Anti-CDH17 antigen-binding domains of the present disclosure can include an antibody or an antigen-binding fragment of an antibody, such as a human antibody, a humanized antibody; a camelid antibody; a chimeric antibody; a recombinant antibody; a heavy chain antibody; a single-domain antibody (e.g., VHH); a single chain antibody (e.g., single chain fragment variable (scFv)); a diabody; a triabody; a tetrabody; a Fab fragment; a F(ab') 2 fragment; an IgD antibody; an IgE antibody; an IgM antibody; an IgGl antibody; an lgG2 antibody; an lgG3 antibody; or an lgG4 antibody, and fragments thereof.

[0404] In some embodiments, an anti-CDH17 antigen-binding domain that binds to CDH17 is a singledomain antibody (also termed as "sdAb"). The single-domain antibodies of the present disclosure can be derived from numerous sources, including but not limited to VHHs, VNARs, or VH domains (naturally occurring or engineered VH domains). VHHs can be generated from camelid heavy chain only antibodies and libraries (e.g., synthetic libraries) thereof. VNARs can be generated from cartilaginous fish heavy chain only antibodies and libraries (e.g., synthetic libraries) thereof. Various methods have been implemented to generate monomeric sdAbs from conventionally heterodimeric VH and VL domains,119323195900v2Attorney Docket No: 260525.000128including interface engineering and selection of specific germline families. In some embodiments, the sdAb of the present invention are human or humanized.

[0405] In some embodiments, an anti-CDH17 single-domain antibody described herein is a VHH fragment (also known as a nanobody). VHH fragments are also referred to as " V-bodies" in the present disclosure. In some embodiments, the VHH is a camelid VHH, a humanized VHH or a camelized VH. In some embodiments, a single-domain antibody described herein is a VH domain. In some embodiments, a single-domain antibody described herein is a naturally occurring VH domain or engineered VH domain.

[0406] The variable domain of an anti-CDH17 antigen-binding domain (e.g., antibody such as a singledomain antibody) of the present disclosure comprises at least three complementarity determining regions (CDRs) which determine its binding specificity. Preferably, in a variable domain, the CDRs are distributed between framework regions (FRs). The variable domain typically contains 4 framework regions interspaced by 3 CDR regions, resulting in the following typical antibody variable domain structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. CDRs and / or FRs of the single domain antibody of the invention may be fragments or derivatives from a naturally occurring antibody variable domain or may be synthetic.

[0407] Sequence identifiers corresponding to exemplary anti-CDH17 VHH antibodies provided herein are listed in Table 1-3. Table 1-3 sets forth the sequence identifiers of amino acid sequences of the complementarity determining regions (CDR1, CDR2 and CDRS), amino acid and DNA sequences of the full-length camelid VHH antibodies, as well as amino acid sequences of corresponding humanized VHH antibodies. In the present disclosure, the suffix "-Hui" in an antibody ID indicates a humanized version of the referenced antibody. Amino acid sequences of additional exemplary anti-CDH17 VHH antibodies and corresponding humanized VHH antibodies are provided in Table 1-4.Table 1-3. Sequence identifiers for exemplary anti-CDH17 VHH antibodiesCDR1 CDR2 CDR3 Non-humanized VHH Humanized VHH Antibody ID Group DNA Amino acid Amino acid sequencesequence sequence C00004 A 2501 2502 2503 2504 2567 2505 C00013 B 2506 2507 2508 2509 2568 2510 C00007 C 2511 2512 2513 2514 2569 2515 C00006 D 2516 2517 2518 2519 2570 2520 C00010 D 2521 2517 2522 2523 2571 2524 C00002 D 2516 2517 2525 2526 2572 2527 C00012 E 2528 2517 2529 2530 2573 2531 C00015 F 3137 3375 3613 2687 3851 2921C00001 F 2532 2533 2534 2535 2574 2536120323195900v2Attorney Docket No: 260525.000128CDR1 CDR2 CDR3 Non-humanized VHH Humanized VHH Antibody ID Group DNA Amino acid Amino acid sequencesequence sequence C00011 G 2537 2538 2539 2540 2575 2541 C00009 H 2542 2543 2544 2545 2576 2546 C00005 1 2547 2548 2549 2550 2577 2551 C00014 1 2552 2553 2554 2555 2578 2556 C00008 J 2557 2558 2559 2560 2579 2561 C00003 K 2562 2563 2564 2565 2580 2566 C00016 M 4107 4108 4109 4119 4127 4123 C00017 N 4110 4111 4112 4120 4128 4124 C00018 0 4113 4114 4115 4121 4129 4125C00019 P 4116 4117 4118 4122 4130 4126 Table 1-4. Sequence identifiers for additional exemplary anti-CDH17 VHH antibodies and humanized VHH antibodiesNon-humanized VHH Humanized VHHGroupAmino Acid SequenceA 2504 2505A 2605 2843A 2606 2844A 2607 2845A 2608 2846A 2609 2847A 2610 2505A 2611 2848A 2612 2505A 2613 2849B 2614 2850B 2509 2510B 2615 2851B 2616 2852B 2617 2853B 2618 2854B 2619 2855B 2620 2856B 2621 2857B 2622 2858B 2623 2859B 2624 2860B 2625 2861B 2626 2862B 2627 2863121323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceC 2514 2515C 2628 2864c 2629 2865c 2630 2866c 2631 2867D 2523 2524D 2519 2520D 2632 2868D 2633 2869D 2634 2870D 2635 2871D 2636 2872D 2637 2873D 2638 2874D 2639 2875D 2640 2876D 2641 2877D 2642 2878D 2643 2879D 2644 2880D 2526 2527D 2645 2881D 2646 2882D 2647 2883D 2648 2884D 2649 2885D 2650 2886D 2651 2887D 2652 2888D 2653 2889D 2654 2890D 2655 2891D 2656 2892D 2657 2893D 2658 2894D 2659 2895D 2660 2896D 2661 2897D 2662 2898D 2663 2899D 2664 2900D 2665 2901D 2666 2902122323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceD 2667 2903D 2668 2877D 2669 2904D 2670 2905D 2671 2906D 2672 2907D 2673 2908D 2674 2909D 2675 2910D 2676 2527D 2677 2911D 2678 2912D 2679 2913D 2680 2914D 2681 2915D 2682 2916D 2683 2917D 2684 2918D 2685 2919D 2686 2920E 2530 2531F 2687 2921F 2535 2536F 2688 2536F 2689 2921F 2690 2922F 2691 2923F 2692 2921F 2693 2924F 2694 2925G 2540 2541G 2695 2926G 2696 2927G 2697 2928H 2545 2546H 2698 2546H 2699 2546H 2700 2546H 2701 25461 2702 29291 2703 29301 2704 29311 2705 2932123323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid Sequence1 2706 29331 2707 29341 2708 29351 2709 29361 2710 29371 2711 29381 2712 29391 2713 29401 2714 29411 2550 25511 2715 29421 2716 29431 2717 29441 2718 29401 2719 29451 2720 29461 2721 25511 2722 29361 2723 29471 2724 29481 2725 29491 2726 29501 2727 29511 2728 29521 2729 29531 2730 29541 2731 29551 2732 25511 2733 29561 2734 29571 2735 29581 2736 29351 2737 29591 2738 29601 2739 29611 2740 29621 2741 29631 2742 29641 2743 29311 2744 29651 2745 29661 2746 29671 2747 2938124323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid Sequence1 2748 29681 2749 29691 2750 29701 2555 25561 2751 29711 2752 29721 2753 29731 2754 29741 2755 29751 2756 29761 2757 29771 2758 29781 2759 29791 2760 29801 2761 29811 2762 29821 2763 25511 2764 29831 2765 29841 2766 29851 2767 29861 2768 29871 2769 29881 2770 29891 2771 29901 2772 29911 2773 29921 2774 29931 2775 29941 2776 29951 2777 29961 2778 29971 2779 29981 2780 29311 2781 29991 2782 30001 2783 30011 2784 30001 2785 30021 2786 30031 2787 30041 2788 30051 2789 3006125323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid Sequence1 2790 29381 2791 3007J 2560 2561J 2792 3008J 2793 3009J 2794 3010J 2795 3011J 2796 3012J 2797 3013J 2798 3013J 2799 2561J 2800 3014J 2801 3015K 2802 3016K 2803 3016K 2804 3017K 2805 3018K 2806 3019K 2807 3020K 2808 3021K 2809 3022K 2810 3023K 2811 3024K 2812 3025K 2813 3026K 2814 3027K 2815 3028K 2816 3029K 2817 3030K 2818 3031K 2819 3032K 2820 3033K 2821 3034K 2822 3035K 2823 3036K 2824 3037K 2825 3038K 2565 2566K 2826 3039K 2827 3040K 2828 3041K 2829 3042K 2830 3043126323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceK 2831 3044K 2832 3045K 2833 3046K 2834 3047K 2835 3019K 2836 3048K 2837 3049K 2838 3050K 2839 3051K 2840 3052K 2841 3053K 2842 3054M 4119 4123M 4143 4172M 4144 4173M 4145 4123N 4120 4124N 4146 4174N 4147 41750 4148 41760 4121 41250 4149 41770 4150 41780 4151 41790 4152 41800 4153 41780 4154 41780 4155 41810 4156 41800 4157 41820 4158 41810 4159 41830 4160 41840 4161 41850 4162 41860 4163 41860 4164 41810 4165 41820 4166 41870 4167 41780 4168 41880 4169 4189P 4122 4126127323195900v2Attorney Docket No: 260525.000128Non-humanized VHH Humanized VHHGroupAmino Acid SequenceP 4170 4190P 4171 4191

[0408] In some embodiments, an anti-CDH17 antigen-binding domain (e.g., antibody such as a singledomain antibody) described herein may comprise a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from (amino acids listed in a pair of brackets represent the possible amino acids at the particular position):a). A(A / L)CLLRFE(S / T)CLEYNRAQY(N / P)Y (SEQ ID NO: 2583);b). AAVR(A / S)GSDWWTTM(R / T)QR(D / H)YD(F / Y) (SEQ ID NO: 2585);c). AARDSR(K / R)GGLFADLN(E / G)YDY (SEQ ID NO: 2588);d). AATG(D / N)(L / S)YRGAYDRP(A / T)EYDY (SEQ ID NO: 2590);e). VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529);f). AAQFSLPVDA(S / T)PLRRY(A / Y)(H / Y) (SEQ ID NO: 2593);g). AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);h). AARNGGYDLNDYAY (SEQ ID NO: 2544);i). NAGG(G / P / A)(A / R)(L / R)GY (SEQ ID NO: 2598);j). NVGGQL(K / L / R)GY (SEQ ID NO: 2601);k). NQGG(Q / S)KGY (SEQ ID NO: 2604);l). AADG(L / P)PY(G / S)(D / S)WFGDQFDV (SEQ ID NO: 4133);m). AFNKWGRLSADL(D / N)DYFR (SEQ ID NO: 4136);n). N(M / T)HRSY(A / D)I(D / N / R / S)FYDN (SEQ ID NO: 4139); ando). RRYDDY(D / G)S (SEQ ID NO: 4142).

[0409] In some embodiments, the CDR3 of the anti-CDH17 antigen-binding domain (e.g., antibody such as a single-domain antibody) described herein comprises an amino acid sequence selected from ALCLLRFETCLEYNRAQYPY (SEQ ID NO: 2503); AAVRSGSDWWTTMTQRHYDF (SEQ ID NO: 2508);AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513); AATGDSYRGAYDRPAEYDY (SEQ ID NO: 2518);AATGNSYRGAYDRPTEYDY (SEQ ID NO: 2522); AATGDLYRGAYDRPAEYDY (SEQ ID NO: 2525);VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529); AAQFSLPVDASPLRRYYY (SEQ ID NO: 2534);AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539); AARNGGYDLNDYAY (SEQ ID NO: 2544); NAGGGRLGY (SEQ ID NO: 2549); NAGGAALGY (SEQ ID NO: 2554); NVGGQLLGY (SEQ ID NO: 2559); NQGGSKGY (SEQ ID NO: 2564); AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613); AADGLPYGDWFGDQFDV (SEQ ID NO: 4109);128323195900v2Attorney Docket No: 260525.000128AFNKWGRLSADLDDYFR (SEQ ID NO: 4112); NMHRSYDISFYDN (SEQ ID NO: 4...

Claims

Attorney Docket No: 260525.000128Claims1. A multispecific antigen-binding protein comprising:a. one or more antigen-binding domains that specifically bind to TNF-related apoptosisinducing ligand receptor 2 (TRAILR2); andb. one or more antigen-binding domains that specifically bind to cadherin 17 (CDH17).

2. The multispecific antigen-binding protein of claim 1, wherein the multispecific antigen-binding protein is bivalent, trivalent, tetravalent, pentavalent, hexavalent, septivalent, or octavalent.

3. The multispecific antigen-binding protein of claim 1 or 2, wherein the multispecific antigen-binding protein comprises at least two, at least three, or at least four anti-TRAILR2 antigen-binding domains, and at least one, at least two, or at least four anti-CDH17 antigen-binding domains.

4. The multispecific antigen-binding protein of any one of claims 1-3, further comprising one or more antigen-binding domains that binds to another antigen.

5. The multispecific antigen-binding protein of claim 4, wherein the other antigen is serum albumin.

6. The multispecific antigen-binding protein of claim 5, wherein the serum albumin is human serum albumin (HSA).

7. The multispecific antigen-binding protein of any one of claims 1-6, which further comprises an immunoglobulin Fc region.

8. A multispecific antigen-binding protein comprising a single polypeptide chain, said single polypeptide chain comprisingi) one or more antigen-binding domains that specifically bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2);ii) one or more antigen-binding domain(s) that specifically bind to cadherin-17 (CDH17); and iii) optionally, one or more antigen-binding domains that bind human serum albumin (HSA); and optionally, wherein each of the antigen-binding domains is operably linked to each of the other antigen-binding domains via a linker.493323195900v2Attorney Docket No: 260525.0001289. The multispecific antigen-binding protein of claim 8, wherein the single polypeptide chain comprises three antigen-binding domains (ABDI- ABD3), four antigen-binding domains (ABDI- ABD4), five antigen-binding domains (ABDI- ABD5), or six antigen-binding domains (ABD1-ABD6).

10. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises three antigen-binding domains, said three antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2; andiii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2.

11. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises four antigen binding domains, said four antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2; andiv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2.

12. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises four antigen binding domains, said four antigen-binding domains comprising, in an N- to C-terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17;ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2; andiv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2.

13. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,494323195900v2Attorney Docket No: 260525.000128ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

14. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

15. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds HSA; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

16. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds HSA.495323195900v2Attorney Docket No: 260525.00012817. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds TRAILR2;ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds CDH17;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2; andv) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2.

18. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises five antigen-binding domains, said five antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds TRAILR2;ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds CDH17; andv) a fifth antigen-binding domain (ABD5) that specifically binds CDH17.

19. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

20. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,496323195900v2Attorney Docket No: 260525.000128ii) a second antigen-binding domain (ABD2) that specifically binds TRAILR2;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds HSA.

21. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds HSA.

22. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds HSA; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

23. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds TRAILR2;497323195900v2Attorney Docket No: 260525.000128iv) a fourth antigen-binding domain (ABD4) that specifically binds HSA;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

24. The multispecific antigen-binding protein of claim 8 or 9, wherein the single polypeptide chain comprises six antigen-binding domains, said six antigen-binding domains comprising, in an N- to C- terminal orientation:i) a first antigen-binding domain (ABDI) that specifically binds CDH17,ii) a second antigen-binding domain (ABD2) that specifically binds CDH17;iii) a third antigen-binding domain (ABD3) that specifically binds HSA;iv) a fourth antigen-binding domain (ABD4) that specifically binds TRAILR2;v) a fifth antigen-binding domain (ABD5) that specifically binds TRAILR2; andvi) a sixth antigen-binding domain (ABD6) that specifically binds TRAILR2.

25. A multispecific antigen-binding protein comprising a first polypeptide chain and a second polypeptide chain, each polypeptide chain comprising:i) one or more antigen-binding domains that specifically bind to TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2),ii) one or more antigen-binding domain that specifically bind to cadherin-17 (CDH17); and iii) an immunoglobulin Fc region; andwherein the first and second polypeptide chains dimerize via the immunoglobulin Fc region to form a dimer, and optionally, wherein each of the antigen-binding domains is operably linked to each of the other antigen-binding domains or the immunoglobulin Fc region via a linker.

26. The multispecific antigen-binding protein of claim 25, wherein the first polypeptide chain and / or the second polypeptide chain each comprises two antigen-binding domains (ABD1-ABD2 or ABD3- ABD4), three antigen-binding domains (ABD1-ABD3 or ABD4-ABD6), or four antigen-binding domains (ABD1-ABD4 or ABD5-ABD8).

27. The multispecific antigen-binding protein of claim 25 or 26, wherein the first polypeptide chain and the second polypeptide chain each comprise two antigen-binding domains (ABD1-ABD2 or ABD3- ABD4), wherein:498323195900v2Attorney Docket No: 260525.000128a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, and (ii) a second antigen-binding domain (ABD2) operably linked to the C-terminus of the first immunoglobulin Fc region; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a third antigen binding domain (ABD3) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, and (ii) a fourth antigen-binding domain (ABD4) operably linked to the C-terminus of the second immunoglobulin Fc region.

28. The multispecific antigen-binding protein of claim 27, wherein ABDI and ABD3 specifically binds to CDH17, and ABD2 and ABD4 specifically bind to TRAILR2.

29. The multispecific antigen-binding protein of claim 27, wherein ABDI and ABD3 specifically bind to TRAILR2, and ABD2 and ABD4 specifically bind to CDH17.

30. The multispecific antigen-binding protein of claim 25 or 26, wherein the first polypeptide chain and the second polypeptide chain each comprise three antigen-binding domains (ABD1-ABD3 or ABD4- ABD6), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, (ii) a second antigen-binding domain (ABD2) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker, and (iii) a third antigen-binding domain (ABD3) operably linked to the second antigen-binding domain (ABD2) via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fourth antigen binding domain (ABD4) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, (ii) a fifth antigen-binding domain (ABD5) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker, and (iii) a sixth antigen-binding domain (ABD6) operably linked to the fifth antigen-binding domain (ABD5) via a linker.

31. The multispecific antigen-binding protein of claim 30, wherein ABDI and ABD4 specifically bind to CDH17, and ABD2, ABD3, ABD5, and ABD6 specifically bind to TRAILR2.499323195900v2Attorney Docket No: 260525.00012832. The multispecific antigen-binding protein of claim 30, wherein ABDI and ABD4 specifically bind to TRAILR2, and ABD2, ABD3, ABD5, and ABD6 specifically bind to CDH17.

33. The multispecific antigen-binding protein of claim 25 or 26, wherein the first polypeptide chain and the second polypeptide chain each comprise three antigen-binding domains (ABD1-ABD3 or ABD4- ABD6), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to a second antigen-binding domain (ABD2) via a linker, (ii) said second antigen binding domain (ABD2) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, and (iii) a third antigen-binding domain (ABD3) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker; andb). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fourth antigen binding domain (ABD4) operably linked to a fifth antigen-binding domain (ABD5) via a linker, (ii) said fifth antigen binding domain (ABD5) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, and (iii) a sixth antigen-binding domain (ABD6) operably linked to the C-terminus of the second immunoglobulin Fc region via a linker.

34. The multispecific antigen-binding protein of claim 33, wherein ABDI, ABD2, ABD4, and ABD5 specifically bind to CDH17, and ABD3 and ABD6 specifically bind to TRAILR2.

35. The multispecific antigen-binding protein of claim 33, wherein ABDI, ABD2, ABD4, and ABD5 specifically bind to TRAILR2, and ABD3 and ABD6 specifically bind to CDH17.

36. The multispecific antigen-binding protein of claim 25 or 26, wherein the first polypeptide chain and the second polypeptide chain each comprise four antigen-binding domains (ABD1-ABD4 or ABD5- ABD8), wherein:a), the first polypeptide chain comprises, in an N- to C-terminal orientation, (i) a first antigen binding domain (ABDI) operably linked to a second antigen-binding domain (ABD2) via a linker, ii) said second antigen binding domain (ABD2) operably linked to the N-terminus of a first immunoglobulin Fc region via a linker, (iii) a third antigen-binding domain (ABD3) operably linked to the C-terminus of the first immunoglobulin Fc region via a linker, and iv) said third antigen-binding domain (ABD3) operably linked to a fourth antigen-binding domain (ABD4) via a linker; and500323195900v2Attorney Docket No: 260525.000128b). the second polypeptide chain comprises, in an N- to C-terminal orientation, (i) a fifth antigen binding domain (ABD5) operably linked to a sixth antigen-binding domain (ABD6) via a linker, ii) said sixth antigen binding domain (ABD6) operably linked to the N-terminus of a second immunoglobulin Fc region via a linker, (iii) a seventh antigen-binding domain (ABD7) operably linked to the C- terminus of the second immunoglobulin Fc region via a linker, and iv) said seventh antigen-binding domain (ABD7) operably linked to an eighth antigen-binding domain (ABD8) via a linker.

37. The multispecific antigen-binding protein of claim 36, wherein ABDI, ABD2, ABD5, and ABD6 specifically bind to CDH17, and ABD3, ABD4, ABD7, and ABD8 specifically bind to TRAILR2.

38. The multispecific antigen-binding protein of claim 36, wherein ABDI, ABD2, ABD5, and ABD6 specifically bind to TRAILR2, and ABD3, ABD4, ABD7, and ABD8 specifically bind to CDH17.

39. The multispecific antigen-binding protein of any of claims 1-38, wherein the one or more anti- TRAILR2 antigen-binding domains comprise a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from:a). (A / V)ASRL(P / V)FNSRSA(I / V)YTDRIYDS (SEQ ID NO: 100);b). AVRRSAWY(S / T)DSIYTVSQYDY (SEQ ID NO: 101);c). NAARSYSR(D / G / N)(G / Y)(E / R)PL(E / K)P(A / D)Y (SEQ ID NO: 102);d). AAASSWSRGG(A / G / I / V)PYGMDY (SEQ ID NO: 103);e). AADS(H / R)FRR(P / Y)(A / T / V)PG(I / Q)QYEY (SEQ ID NO: 104);f). NAA(K / R)SYHRDY(K / S)PL(K / S)(G / P)DY (SEQ ID NO: 105);g). AAAPSFGM(M / R / T)(I / N)PESYVHS (SEQ ID NO: 106);h). AANRGIMSMRLSRYDD (SEQ ID NO: 49);i). T(A / V)GP(A / T)MSYSRGGEF (SEQ ID NO: 107);j). (A / V)ADRGAISRSGAGM(D / N)Y (SEQ ID NO: 108);k). TAGP(A / S)IS(L / Y)SRGGEY (SEQ ID NO: 109);l). A(A / T)NGWGLDP(S / T)TYH(Y / D) (SEQ ID NO: 110);m). SAGWTRRIFQY (SEQ ID NO: 88);n). TAGQSISLSQGGE(H / Y) (SEQ ID NO: 111);o). KAGIRGE(T / V)Y (SEQ ID NO: 112);p). RAYNDGGEY (SEQ ID NO: 2191);501323195900v2Attorney Docket No: 260525.000128q). HS(N / R)WYNL (SEQ ID NO: 2208); andr). (F / M / N)T(A / S)DY,wherein one or more non-alanine residues in the CDR3 sequence are optionally replaced with an alanine, and / or one or more alanine residues in the CDR3 sequence are optionally replaced with a glycine.

40. The multispecific antigen-binding protein of claim 39, wherein the CDR3 of the one or more anti- TRAILR2 antigen-binding domains comprises an amino acid sequence selected from:a). (A / G)(A / G)(A / G)(A / S)(A / S)(A / W)(A / S)(A / R)(A / G)(A / G)(A / G / I / V)(A / P)(A / Y)(A / G)(A / M)(A / D) (A / Y);b). (A / T)(A / G / V)(A / G)(A / P)(A / T)(A / M)(A / S)(A / Y)(A / S)(A / R)(A / G)(A / G)(A / E)(A / F); c). (A / R)(A / G)(A / Y)(A / N)(A / D)(A / G)(A / G)(A / E)(A / Y);d). (A / H)(A / S)(A / N / R)(A / W)(A / Y)(A / N)(A / L); ande). (A / F / M / N)(A / T)(A / G / S)(A / D)Y.

41. The multispecific antigen-binding protein of claim 39 or 40, wherein the CDR3 of the one or more anti-TRAILR2 antigen-binding domains comprises an amino acid sequence selected from:VASRLPFNSRSAIYTDRIYDS (SEQ ID NO: 3); AVRRSAWYSDSIYTVSQYDY (SEQ ID NO: 8);NAARSYSRDYEPLKPDY (SEQ ID NO: 13); NAARSYSRNYEPLKPDY (SEQ ID NO: 16);NAARSYSRGGEPLKPDY (SEQ ID NO: 20); NAARSYSRGGRPLEPAY (SEQ ID NO: 25);AAASSWSRGGVPYGMDY (SEQ ID NO: 30); AADSHFRRYTPGQQYEY (SEQ ID NO: 35);NAAKSYHRDYSPLSPDY (SEQ ID NO: 40); AAAPSFGMRNPESYVHS (SEQ ID NO: 44);AANRGIMSMRLSRYDD (SEQ ID NO: 49); TAGPTMSYSRGGEF (SEQ ID NO: 54); VADRGAISRSGAGMDY (SEQ ID NO: 64); AADRGAISRSGAGMDY (SEQ ID NO: 69); TAGPAISLSRGGEY (SEQ ID NO: 74);TAGPSISYSRGGEY (SEQ ID NO: 78); AANGWGLDPTTYHY (SEQ ID NO: 83); SAGWTRRIFQY (SEQ ID NO: 88); TAGQSISLSQGGEY (SEQ ID NO: 92); KAGIRGEVY (SEQ ID NO: 97); RAYNDGGEY (SEQ ID NO: 2191); HSRWYNL (SEQ ID NO: 2197); FTADY (SEQ ID NO: 2203); GAASSWSRGGVPYGMDY (SEQ ID NO: 2298); AGASSWSRGGVPYGMDY (SEQ ID NO: 2299); AAGSSWSRGGVPYGMDY (SEQ ID NO: 2300); AAAASWSRGGVPYGMDY (SEQ ID NO: 2301); AAASAWSRGGVPYGMDY (SEQ ID NO: 2302);AAASSASRGGVPYGMDY (SEQ ID NO: 2303); AAASSWARGGVPYGMDY (SEQ ID NO: 2304);AAASSWSAGGVPYGMDY (SEQ ID NO: 2305); AAASSWSRAGVPYGMDY (SEQ ID NO: 2306);AAASSWSRGAVPYGMDY (SEQ ID NO: 2307); AAASSWSRGGAPYGMDY (SEQ ID NO: 2308);502323195900v2Attorney Docket No: 260525.000128AAASSWSRGGVAYGMDY (SEQ ID NO: 2309); AAASSWSRGGVPAGMDY (SEQ ID NO: 2310);AAASSWSRGGVPYAMDY (SEQ ID NO: 2311); AAASSWSRGGVPYGADY (SEQ ID NO: 2312);AAASSWSRGGVPYGMAY (SEQ ID NO: 2313); AAASSWSRGGVPYGMDA (SEQ ID NO: 2314);AAGPTMSYSRGGEF (SEQ ID NO: 2315); TGGPTMSYSRGGEF (SEQ ID NO: 2316); TAAPTMSYSRGGEF (SEQ ID NO: 2317); TAGATMSYSRGGEF (SEQ ID NO: 2318); TAGPAMSYSRGGEF (SEQ ID NO: 2319); TAGPTASYSRGGEF (SEQ ID NO: 2320); TAGPTMAYSRGGEF (SEQ ID NO: 2321); TAGPTMSASRGGEF (SEQ ID NO: 2322); TAGPTMSYARGGEF (SEQ ID NO: 2323); TAGPTMSYSAGGEF (SEQ ID NO: 2324); TAGPTMSYSRAGEF (SEQ ID NO: 2325); TAGPTMSYSRGAEF (SEQ ID NO: 2326); TAGPTMSYSRGGAF (SEQ ID NO: 2327); TAGPTMSYSRGGEA (SEQ ID NO: 2328); AAYNDGGEY (SEQ ID NO: 2329);RGYNDGGEY (SEQ ID NO: 2330); RAANDGGEY (SEQ ID NO: 2331); RAYADGGEY (SEQ ID NO: 2332); RAYNAGGEY (SEQ ID NO: 2333); RAYNDAGEY (SEQ ID NO: 2334); RAYNDGAEY (SEQ ID NO: 2335); RAYNDGGAY (SEQ ID NO: 2336); RAYNDGGEA (SEQ ID NO: 2337); ASRWYNL (SEQ ID NO: 2338); HARWYNL (SEQ ID NO: 2339); HSAWYNL (SEQ ID NO: 2340); HSRAYNL (SEQ ID NO: 2341); HSRWANL (SEQ ID NO: 2342); HSRWYAL (SEQ ID NO: 2343); HSRWYNA (SEQ ID NO: 2344); ATADY (SEQ ID NO: 2345); FAADY (SEQ ID NO: 2346); FTGDY (SEQ ID NO: 2347); and FTAAY (SEQ ID NO: 2348).

42. The multispecific antigen-binding protein of any of claims 1-41, wherein the one or more anti- TRAILR2 antigen-binding domains further comprise a CDR1 comprising an amino acid sequence selected from:a). GRTFSSNL (SEQ ID NO: 1);b). GGT(F / L)(A / S)N(D / N)G (SEQ ID NO: 113);c). GRTL(D / N / S)(A / D / E)Y(A / G) (SEQ ID NO: 114);d). GRTFS(N / S)YA (SEQ ID NO: 115);e) GRDFSNYV (SEQ ID NO: 33);f). G(L / R)(I / S)FS(D / S)YA (SEQ ID NO: 116);g). GR(A / T)FSTLA (SEQ ID NO: 117);h). GRTFSSDI (SEQ ID NO: 47);i). GRSFGD(D / F / Y)A (SEQ ID NO: 118);j). G(G / R)TLSNYA (SEQ ID NO: 119);k). GRSFGAQGMEG (SEQ ID NO: 72);l). GFTLDLGAYA (SEQ ID NO: 81);m). GFTFGALA (SEQ ID NO: 86);503323195900v2Attorney Docket No: 260525.000128n). GFTLS(G / S)YA (SEQ ID NO: 120);o). GSIFGGYN (SEQ ID NO: 2189);p). G(G / S)NFRILS (SEQ ID NO: 2209); andq). G(F / L)(A / T)F(R / S)(R / S)YA (SEQ ID NO: 2212).

43. The multispecific antigen-binding protein of claim 42, wherein the CDR1 of the one or more anti- TRAILR2 antigen-binding domains comprises an amino acid sequence selected from GRTFSSNL (SEQ ID NO: 1); GGTLANNG (SEQ ID NO: 6); GRTLDAYG (SEQ ID NO: 11); GRTLSDYA (SEQ ID NO: 23); GRTFSSYA (SEQ ID NO: 28); GRDFSNYV (SEQ ID NO: 33); GRSFSSYA (SEQ ID NO: 38); GRTFSTLA (SEQ ID NO: 43); GRTFSSDI (SEQ ID NO: 47); GRSFGDFA (SEQ ID NO: 52); GGTLSNYA (SEQ ID NO: 62); GRTLSNYA (SEQ ID NO: 67); GRSFGAQGMEG (SEQ ID NO: 72); GFTLDLGAYA (SEQ ID NO: 81);GFTFGALA (SEQ ID NO: 86); GFTLSGYA (SEQ ID NO: 95); GSIFGGYN (SEQ ID NO: 2189); GSNFRILS (SEQ ID NO: 2195); and GFTFSRYA (SEQ ID NO: 2201).

44. The multispecific antigen-binding protein of any one of claims 1-43, wherein the one or more anti- TRAILR2 antigen-binding domains further comprise a CDR2 comprising an amino acid sequence selected from:a). VSWNGAST (SEQ ID NO: 2);b). DHR(S / T)GT (SEQ ID NO: 121);c). I(N / S)W(N / S / T)G(T / V)(D / G)T (SEQ ID NO: 122);d). LNW(N / S)G(D / E)ST (SEQ ID NO: 123);e). INWAD(E / T)T (SEQ ID NO: 124);f). INWSGG(S / T)T (SEQ ID NO: 125);g). ISWSDMSA (SEQ ID NO: 48);h). I(N / R)W(A / D / T)G(D / N)T (SEQ ID NO: 126);i). ISQ(S / T)S(D / S)T (SEQ ID NO: 127);j). (I / M)KWTGNT (SEQ ID NO: 128);k). ISN(S / T)GTTT (SEQ ID NO: 129);l). ISNDGEHI (SEQ ID NO: 87);m). ISWNGDIT (SEQ ID NO: 91);n). IT(G / S)(A / S)G(G / S)(N / S)T (SEQ ID NO: 130);o). IFISGN(D / N) (SEQ ID NO: 2207);504323195900v2Attorney Docket No: 260525.000128p). (I / L)T(K / M / S)D(D / G)TT (SEQ ID NO: 2210); andq). ISS(A / G / S)(G / S)G(I / Y)(I / T / V) (SEQ ID NO: 2213).

45. The multispecific antigen-binding protein of claim 44, wherein the CDR2 of the one or more anti- TRAILR2 antigen-binding domains comprises an amino acid sequence selected from VSWNGAST (SEQ ID NO: 2); DHRSGT (SEQ ID NO: 7); ISWTGVDT (SEQ ID NO: 12); ISWTGTDT (SEQ ID NO: 19); ISWSGVDT (SEQ ID NO: 24); LNWSGEST (SEQ ID NO: 29); INWADET (SEQ ID NO: 34); INWSGGST (SEQ ID NO: 39); ISWSDMSA (SEQ ID NO: 48); IRWTGDT (SEQ ID NO: 53); ISQTSST (SEQ ID NO: 63);ISQSSDT(SEQ ID NO: 68); MKWTGNT (SEQ ID NO: 73); IKWTGNT (SEQ ID NO: 77); ISNTGTTT (SEQ ID NO: 82); ISNDGEHI (SEQ ID NO: 87); ISWNGDIT (SEQ ID NO: 91); ITSAGGST (SEQ ID NO: 96); IFISGNN (SEQ ID NO: 2190); ITSDDTT (SEQ ID NO: 2196); and ISSAGGYI (SEQ ID NO: 2202).

46. The multispecific antigen-binding protein of any one of claims 1-45, wherein the one or more anti- TRAILR2 antigen-binding domains comprise a CDR1 comprising an amino acid sequence selected from SEQ ID NOs: 1, 6, 11, 23, 28, 33, 38, 43, 47, 52, 62, 67, 72, 81, 86, 95, 762-1084, 2189, 2195, 2201, and 2252-2261; a CDR2 comprising an amino acid sequence selected from SEQ ID NOs: 2, 7, 12, 19, 24, 29, 34, 39, 48, 53, 63, 68, 73, 77, 82, 87, 91, 96, 1085-1407, 2190, 2196, 2202, and 2262- 2271; and / or a CDR3 comprising an amino acid sequence selected from SEQ ID NOs: 3, 8, 13, 16, 20, 25, 30, 35, 40, 44, 49, 54, 64, 69, 74, 78, 83, 88, 92, 97, 1408-1730, 2191, 2197, 2203, 2272-2277, and 2298-2348.

47. The multispecific antigen-binding protein of any one of claims 1-46, wherein the one or more anti- TRAILR2 antigen-binding domains comprise:i). a CDR1 comprising an amino acid sequence of GRTFSSNL (SEQ ID NO: 1), a CDR2 comprising an amino acid sequence of VSWNGAST (SEQ ID NO: 2), and a CDR3 comprising an amino acid sequence of VASRLPFNSRSAIYTDRIYDS (SEQ ID NO: 3);ii). a CDR1 comprising an amino acid sequence of GGTLANNG (SEQ ID NO: 6), a CDR2 comprising an amino acid sequence of DHRSGT (SEQ ID NO: 7), and a CDR3 comprising an amino acid sequence of AVRRSAWYSDSIYTVSQYDY (SEQ ID NO: 8);iii). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), and a CDR3 comprising an amino acid sequence of NAARSYSRDYEPLKPDY (SEQ ID NO: 13);505323195900v2Attorney Docket No: 260525.000128iv). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), and a CDR3 comprising an amino acid sequence of NAARSYSRNYEPLKPDY (SEQ ID NO: 16);v). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGTDT (SEQ ID NO: 19), and a CDR3 comprising an amino acid sequence of NAARSYSRGGEPLKPDY (SEQ ID NO: 20);vi). a CDR1 comprising an amino acid sequence of GRTLSDYA (SEQ ID NO: 23), a CDR2 comprising an amino acid sequence of ISWSGVDT (SEQ ID NO: 24), and a CDR3 comprising an amino acid sequence of NAARSYSRGGRPLEPAY (SEQ ID NO: 25);vii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);viii). a CDR1 comprising an amino acid sequence of GRDFSNYV (SEQ ID NO: 33), a CDR2 comprising an amino acid sequence of INWADET (SEQ ID NO: 34), and a CDR3 comprising an amino acid sequence of AADSHFRRYTPGQQYEY (SEQ ID NO: 35);ix). a CDR1 comprising an amino acid sequence of GRSFSSYA (SEQ ID NO: 38), a CDR2 comprising an amino acid sequence of INWSGGST (SEQ ID NO: 39), and a CDR3 comprising an amino acid sequence of NAAKSYHRDYSPLSPDY (SEQ ID NO: 40);x). a CDR1 comprising an amino acid sequence of GRTFSTLA (SEQ ID NO: 43), a CDR2 comprising an amino acid sequence of INWSGGST (SEQ ID NO: 39), and a CDR3 comprising an amino acid sequence of AAAPSFGMRNPESYVHS (SEQ ID NO: 44);xi). a CDR1 comprising an amino acid sequence of GRTFSSDI (SEQ ID NO: 47), a CDR2 comprising an amino acid sequence of ISWSDMSA (SEQ ID NO: 48), and a CDR3 comprising an amino acid sequence of AANRGIMSMRLSRYDD (SEQ ID NO: 49);xii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);xiii). a CDR1 comprising an amino acid sequence of GGTLSNYA (SEQ ID NO: 62), a CDR2 comprising an amino acid sequence of ISQTSST (SEQ ID NO: 63), and a CDR3 comprising an amino acid sequence of VADRGAISRSGAGMDY (SEQ ID NO: 64);506323195900v2Attorney Docket No: 260525.000128xiv). a CDR1 comprising an amino acid sequence of GRTLSNYA (SEQ ID NO: 67), a CDR2 comprising an amino acid sequence of ISQSSDT (SEQ ID NO: 68), and a CDR3 comprising an amino acid sequence of AADRGAISRSGAGMDY (SEQ ID NO: 69);xv). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of MKWTGNT (SEQ ID NO: 73), and a CDR3 comprising an amino acid sequence of TAGPAISLSRGGEY (SEQ ID NO: 74);xvi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78);xvii). a CDR1 comprising an amino acid sequence of GFTLDLGAYA (SEQ ID NO: 81), a CDR2 comprising an amino acid sequence of ISNTGTTT (SEQ ID NO: 82), and a CDR3 comprising an amino acid sequence of AANGWGLDPTTYHY (SEQ ID NO: 83);xviii). a CDR1 comprising an amino acid sequence of GFTFGALA (SEQ ID NO: 86), a CDR2 comprising an amino acid sequence of ISNDGEHI (SEQ ID NO: 87), and a CDR3 comprising an amino acid sequence of SAGWTRRIFQY (SEQ ID NO: 88);xix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92);xx). a CDR1 comprising an amino acid sequence of GFTLSGYA (SEQ ID NO: 95), a CDR2 comprising an amino acid sequence of ITSAGGST (SEQ ID NO: 96), and a CDR3 comprising an amino acid sequence of KAGIRGEVY (SEQ ID NO: 97);xxi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);xxii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);xxiii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);507323195900v2Attorney Docket No: 260525.000128xxiv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of GAASSWSRGGVPYGMDY (SEQ ID NO: 2298);xxv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AGASSWSRGGVPYGMDY (SEQ ID NO: 2299);xxvi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAGSSWSRGGVPYGMDY (SEQ ID NO: 2300);xxvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAAASWSRGGVPYGMDY (SEQ ID NO: 2301);xxviii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASAWSRGGVPYGMDY (SEQ ID NO: 2302);xxix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSASRGGVPYGMDY (SEQ ID NO: 2303);xxx). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWARGGVPYGMDY (SEQ ID NO: 2304);xxxi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSAGGVPYGMDY (SEQ ID NO: 2305);xxxii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRAGVPYGMDY (SEQ ID NO: 2306);xxxiii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGAVPYGMDY (SEQ ID NO: 2307);508323195900v2Attorney Docket No: 260525.000128xxxiv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGAPYGMDY (SEQ ID NO: 2308);xxxv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVAYGMDY (SEQ ID NO: 2309);xxxvi). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPAGMDY (SEQ ID NO: 2310);xxxvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYAMDY (SEQ ID NO: 2311);xxxviii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGADY (SEQ ID NO: 2312);xxxix). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMAY (SEQ ID NO: 2313);xl). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDA (SEQ ID NO: 2314);xli). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of AAGPTMSYSRGGEF (SEQ ID NO: 2315);xlii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TGGPTMSYSRGGEF (SEQ ID NO: 2316);xliii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAAPTMSYSRGGEF (SEQ ID NO: 2317);509323195900v2Attorney Docket No: 260525.000128xliv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGATMSYSRGGEF (SEQ ID NO: 2318);xlv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPAMSYSRGGEF (SEQ ID NO: 2319);xlvi). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTASYSRGGEF (SEQ ID NO: 2320);xlvii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMAYSRGGEF (SEQ ID NO: 2321);xlviii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSASRGGEF (SEQ ID NO: 2322);xlix). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYARGGEF (SEQ ID NO: 2323);I), a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSAGGEF (SEQ ID NO: 2324);li). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRAGEF (SEQ ID NO: 2325);lii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGAEF (SEQ ID NO: 2326);liii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGAF (SEQ ID NO: 2327);510323195900v2Attorney Docket No: 260525.000128liv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEA (SEQ ID NO: 2328);Iv). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of AAYNDGGEY (SEQ ID NO: 2329);Ivi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RGYNDGGEY (SEQ ID NO: 2330);Ivii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAANDGGEY (SEQ ID NO: 2331);Iviii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYADGGEY (SEQ ID NO: 2332);lix). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNAGGEY (SEQ ID NO: 2333);lx), a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDAGEY (SEQ ID NO: 2334);Ixi). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGAEY (SEQ ID NO: 2335);Ixii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGAY (SEQ ID NO: 2336);Ixiii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEA (SEQ ID NO: 2337);511323195900v2Attorney Docket No: 260525.000128Ixiv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of ASRWYNL (SEQ ID NO: 2338);Ixv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HARWYNL (SEQ ID NO: 2339);Ixvi). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSAWYNL (SEQ ID NO: 2340);Ixvii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRAYNL (SEQ ID NO: 2341);Ixviii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWANL (SEQ ID NO: 2342);Ixix). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYAL (SEQ ID NO: 2343);Ixx). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNA (SEQ ID NO: 2344);Ixxi). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of ATADY (SEQ ID NO: 2345);Ixxii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FAADY (SEQ ID NO: 2346);Ixxiii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTGDY (SEQ ID NO: 2347); or512323195900v2Attorney Docket No: 260525.000128Ixxiv). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTAAY (SEQ ID NO: 2348).

48. The multispecific antigen-binding protein of any one of claim 1-47, wherein the one or more anti- TRAILR2 antigen-binding domains comprise:i). a CDR1 comprising an amino acid sequence of GRTLDAYG (SEQ ID NO: 11), a CDR2 comprising an amino acid sequence of ISWTGVDT (SEQ ID NO: 12), and a CDR3 comprising an amino acid sequence of NAARSYSRDYEPLKPDY (SEQ ID NO: 13);ii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRAGEF (SEQ ID NO: 2325);iii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYARGGEF (SEQ ID NO: 2323);iv). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);v). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);vi). a CDR1 comprising an amino acid sequence of GFTLSGYA (SEQ ID NO: 95), a CDR2 comprising an amino acid sequence of ITSAGGST (SEQ ID NO: 96), and a CDR3 comprising an amino acid sequence of KAGIRGEVY (SEQ ID NO: 97);vii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);viii). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);513323195900v2Attorney Docket No: 260525.000128ix). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);x). a CDR1 comprising an amino acid sequence of GLPFRSYA (SEQ ID NO: 2260), a CDR2 comprising an amino acid sequence of ISSGGGIV (SEQ ID NO: 2271), and a CDR3 comprising an amino acid sequence of FASDY (SEQ ID NO: 2276);xi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of MKWTGNT (SEQ ID NO: 73), and a CDR3 comprising an amino acid sequence of TAGPAISLSRGGEY (SEQ ID NO: 74);xii). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orxiii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92).

49. The multispecific antigen-binding protein of any one of claims 1-48, wherein the one or more anti- TRAILR2 antigen-binding domains comprise:i). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of LNWSGEST (SEQ ID NO: 29), and a CDR3 comprising an amino acid sequence of AAASSWSRGGVPYGMDY (SEQ ID NO: 30);ii). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);iii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);iv). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);514323195900v2Attorney Docket No: 260525.000128v). a CDR1 comprising an amino acid sequence of GFTFSRYA (SEQ ID NO: 2201), a CDR2 comprising an amino acid sequence of ISSAGGYI (SEQ ID NO: 2202), and a CDR3 comprising an amino acid sequence of FTADY (SEQ ID NO: 2203);vi). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orvii). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92).

50. The multispecific antigen-binding protein of any one of claims 1-49, wherein the one or more anti- TRAILR2 antigen-binding domains comprise at least one anti-TRAILR2 single-domain antibody.

51. The multispecific antigen-binding protein of claim 50, wherein the anti-TRAILR2 single-domain antibody is an anti-TRAILR2 VHH, an anti-TRAILR2 VNAR, or an anti-TRAILR2 VH domain.

52. The multispecific antigen-binding protein of claim 50, wherein the anti-TRAILR2 single-domain antibody is an anti-TRAILR2 VHH.

53. The multispecific antigen-binding protein of claim 52, wherein the anti-TRAILR2 VHH is a camelid anti-TRAILR2 VHH.

54. The multispecific antigen-binding protein of claim 53, wherein the camelid anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 4, 9, 14, 17, 21, 26, 31, 36, 41, 45, 50, 55, 65, 70, 75, 79, 84, 89, 93, 98, 146-468, 2192, 2198, 2204, and 2214-2233, or an amino acid sequence having at least 75% identity thereto.

55. The multispecific antigen-binding protein of claim 54, wherein the camelid anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 14, 31, 55, 2192, 2198, and 2204, or an amino acid sequence having at least 75% identity thereto.515323195900v2Attorney Docket No: 260525.00012856. The multispecific antigen-binding protein of claim 52, wherein the anti-TRAILR2 VHH is a humanized anti-TRAILR2 VHH.

57. The multispecific antigen-binding protein of claim 56, wherein the humanized anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 5, 10, 15, 18, 22, 27, 32, 37, 42, 46, 51, 56, 66, 71, 76, 80, 85, 90, 94, 99, 469-761, 2194, 2200, 2206, 2234-2251, 2354-2404, 4662-4677, and 4692-4697, or an amino acid sequence having at least 75% identity thereto.

58. The multispecific antigen-binding protein of claim 57, wherein the humanized anti-TRAILR2 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 32, 56, 76, 80, 94, 99, 2194, 2200, 2206, 2250, 2379, 2381, 4662-4677, and 4692-4697, or an amino acid sequence having at least 75% identity thereto59. The multispecific antibody of any of claims 1-58, wherein the one or more anti-TRAILR2 antigenbinding domains that specifically binds to human TRAILR2.

60. The multispecific antigen-binding protein of claim 59, wherein the one or more anti-TRAILR2 antigen-binding domains bind to human TRAILR2 with a KDof less than about 3xl0"7M.

61. The multispecific antigen-binding protein of claim 60, wherein the one or more anti-TRAILR2 antigen-binding domains bind to human TRAILR2 with a KDof about lxlO-10to 5xl0“8M.

62. The multispecific antigen-binding protein of any one of claims 1-61, wherein the one or more anti- TRAILR2 antigen-binding domains bind to cyno TRAILR2.

63. The multispecific antigen-binding protein of claim 62, wherein the one or more anti-TRAILR2 antigen-binding domains bind to cyno TRAILR2 with a KDof less than about 3xl0"7M.

64. The multispecific antigen-binding protein of claim 63, wherein the one or more anti-TRAILR2 antigen-binding domains bind to cyno TRAILR2 with a KDof about lxlO"9to lxlO"7M.516323195900v2Attorney Docket No: 260525.00012865. The multispecific antigen-binding protein of any one of claims 1-64, wherein the one or more anti- TRAILR2 antigen-binding domains do not block binding of TNF-related apoptosis-inducing ligand (TRAIL) to TRAILR2.

66. The multispecific antigen-binding protein of any one of claims 1-64, wherein the one or more anti- TRAILR2 antigen-binding domains block binding of TRAIL to TRAILR2.

67. The multispecific antigen-binding protein of any one of claims 1-66, wherein the one or more anti- TRAILR2 antigen-binding domains do not bind specifically to TRAILR1, TRAILR3 and / or TRAILR4.

68. The multispecific antigen-binding protein of any one of claims 1-66, wherein the one or more anti- TRAILR2 antigen-binding domains bind cysteine-rich domain (CRD) domain 1 (CRD1) and / or CRD2 of TRAILR2.

69. The multispecific antigen-binding protein of any one of claims 1-68, wherein the one or more anti- TRAILR2 antigen-binding domains bind CRD1 of TRAILR2.

70. The multispecific antigen-binding protein of any one of claims 1-68, wherein the one or more anti- TRAILR2 antigen-binding domains bind CRD2 of TRAILR2.

71. The multispecific antigen-binding protein of any one of claims 1-68, wherein the one or more anti- TRAILR2 antigen-binding domains bind CRD1 and CRD2 of TRAILR2.

72. The multispecific antigen-binding protein of any one of claims 1-66, wherein the one or more anti- TRAILR2 antigen-binding domains bind CRD2 and CRD3 of TRAILR2.

73. The multispecific antigen-binding protein of any one of claims 1-72, wherein the one or more anti- TRAILR2 antigen-binding domains bind to the same epitope on TRAILR2.

74. The multispecific antigen-binding protein of any one of claims 1-72, wherein the one or more anti- TRAILR2 antigen-binding domains bind to different epitopes on TRAILR2.517323195900v2Attorney Docket No: 260525.00012875. The multispecific antigen-binding protein of any one of claims 1-74, wherein the one or more anti- CDH17 antigen-binding domains comprise a complementarity determining region 3 (CDR3) comprising an amino acid sequence selected from:a) A(A / L)CLLRFE(S / T)CLEYNRAQY(N / P)Y (SEQ ID NO: 2583);b) AAVR(A / S)GSDWWTTM(R / T)QR(D / H)YD(F / Y) (SEQ ID NO: 2585);c) AARDSR(K / R)GGLFADLN(E / G)YDY (SEQ ID NO: 2588);d) AATG(D / N)(L / S)YRGAYDRP(A / T)EYDY (SEQ ID NO: 2590);e) VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529);f) AAQFSLPVDA(S / T)PLRRY(A / Y)(H / Y) (SEQ ID NO: 2593);g) AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);h) AARNGGYDLNDYAY (SEQ ID NO: 2544);i) NAGG(G / P / A)(A / R)(L / R)GY (SEQ ID NO: 2598);j) NVGGQL(K / L / R)GY (SEQ ID NO: 2601);k) NQGG(Q / S)KGY (SEQ ID NO: 2604);l) AADG(L / P)PY(G / S)(D / S)WFGDQFDV (SEQ ID NO: 4133);m) AFNKWGRLSADL(D / N)DYFR (SEQ ID NO: 4136);n) N(M / T)HRSY(A / D)I(D / N / R / S)FYDN (SEQ ID NO: 4139); ando) RRYDDY(D / G)S (SEQ ID NO: 4142),wherein one or more non-alanine residues in the CDR3 sequence are optionally replaced with an alanine, and / or one or more alanine residues in the CDR3 sequence are optionally replaced with a glycine.

76. The multispecific antigen-binding protein of claim 75, wherein the CDR3 of the one or more anti- CDH17 antigen-binding domains comprises an amino acid sequence selected from:ALCLLRFETCLEYNRAQYPY (SEQ ID NO: 2503); AAVRSGSDWWTTMTQRHYDF (SEQ ID NO: 2508); AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513); AATGDSYRGAYDRPAEYDY (SEQ ID NO: 2518);AATGNSYRGAYDRPTEYDY (SEQ ID NO: 2522); AATGDLYRGAYDRPAEYDY (SEQ ID NO: 2525);VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529); AAQFSLPVDASPLRRYYY (SEQ ID NO: 2534);AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539); AARNGGYDLNDYAY (SEQ ID NO: 2544); NAGGGRLGY (SEQ ID NO: 2549); NAGGAALGY (SEQ ID NO: 2554); NVGGQLLGY (SEQ ID NO: 2559); NQGGSKGY (SEQ ID NO: 2564); AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613); AADGLPYGDWFGDQFDV (SEQ ID NO:518323195900v2Attorney Docket No: 260525.0001284109); AFNKWGRLSADLDDYFR (SEQ ID NO: 4112); NMHRSYDISFYDN (SEQ ID NO: 4115); and RRYDDYGS (SEQ ID NO: 4118).

77. The multispecific antigen-binding protein of any one of claims 1-76, wherein the one or more anti- CDH17 antigen-binding domains further comprise a CDR1 comprising an amino acid sequence selected from:a). GFTLSN(T / Y)N (SEQ ID NO: 2581);b). GSPLDYYA (SEQ ID NO: 2506);c). RL(A / N / T)(F / S)(N / S)(R / S)(S / T)T;d). GRTFS(E / T)PI (SEQ ID NO: 2589);e). GRTFSSPI (SEQ ID NO: 2528);f). TRTF(D / N)MYA (SEQ ID NO: 2591);g). GRTF(D / S)S(L / Y)(L / V) (SEQ ID NO: 2594);h). GRTDSILN (SEQ ID NO: 2542);i). G(I / M)RFS(S / Q)YA (SEQ ID NO: 2596);j). GIRFS(A / S)YA (SEQ ID NO: 2599);k). GS(I / R)FS(R / S)(W / Y)A (SEQ ID NO: 2602);l). G(G / R)TASEYG (SEQ ID NO: 4131);m). EQTMTGF(T / W) (SEQ ID NO: 4134);n). GLRFS(S / N)YA (SEQ ID NO: 4137); ando). (G / R)GT(F / V)SGYA (SEQ ID NO: 4140).

78. The multispecific antigen-binding protein of claim 77, wherein the CDR1 of the one or more anti- CDH17 antigen-binding domains comprises an amino acid sequence selected from GFTLSNYN (SEQ ID NO: 2501); GSPLDYYA (SEQ ID NO: 2506); RLNFSRTT (SEQ ID NO: 2511); GRTFSEPI (SEQ ID NO: 2516); GRTFSTPI (SEQ ID NO: 2521); GRTFSEPI (SEQ ID NO: 2516); GRTFSSPI (SEQ ID NO: 2528); TRTFNMYA (SEQ ID NO: 2532); GRTFSSYL (SEQ ID NO: 2537); GRTDSILN (SEQ ID NO: 2542); GIRFSSYA (SEQ ID NO: 2547); GMRFSQYA (SEQ ID NO: 2552); GIRFSAYA (SEQ ID NO: 2557); GSRFSSYA (SEQ ID NO: 2562); TRTFDMYA (SEQ ID NO: 3137); GGTASEYG (SEQ ID NO: 4107); EQTMTGFW (SEQ ID NO: 4110); GLRFSNYA (SEQ ID NO: 4113); and GGTVSGYA (SEQ ID NO: 4116).519323195900v2Attorney Docket No: 260525.00012879. The multispecific antigen-binding protein of any one of claims 1-78, wherein the one or more anti- CDH17 antigen-binding domains further comprise a CDR2 comprising an amino acid sequence selected from:a) ( F / IJSRGGRT (SEQ ID NO: 2582);b) ISTSGR(C / S)T (SEQ ID NO: 2584);c) SGW(A / S)R(G / T)RT (SEQ ID NO: 2587);d) LISTGGST (SEQ ID NO: 2517);e) l(N / S)RSG(A / T)NT (SEQ ID NO: 2592);f) ISWN(D / G)RST (SEQ ID NO: 2595);g) ISWFRGET (SEQ ID NO: 2543);h) l(F / T)(l / K / N / S)(D / G)(G / Y)(R / S / T)T;i) MT(A / N / T)GGMT (SEQ ID NO: 2600);j) IT(N / S)GG(G / R / S)T (SEQ ID NO: 2603);k) ISTSGGVT (SEQ ID NO: 4108);l) ISASGSRV (SEQ ID NO: 4111);m) IT(N / K)GG(I / N / S)T (SEQ ID NO: 4138); andn) INSGGPT (SEQ ID NO: 4117).

80. The multispecific antigen-binding protein of claim 79, wherein the CDR2 of the one or more anti- CDH17 antigen-binding domains comprises an amino acid sequence selected ISRGGRT (SEQ ID NO: 2502); ISTSGRCT (SEQ ID NO: 2507); SGWARGRT (SEQ ID NO: 2512); LISTGGST (SEQ ID NO: 2517); ISRSGTNT (SEQ ID NO: 2533); ISWNDRST (SEQ ID NO: 2538); ISWFRGET (SEQ ID NO: 2543); ITSGYRT (SEQ ID NO: 2548); IFKDGTT (SEQ ID NO: 2553); MTAGGMT (SEQ ID NO: 2558); ITSGGRT (SEQ ID NO: 2563); INRSGANT (SEQ ID NO: 3375); ISTSGGVT (SEQ ID NO: 4108); ISASGSRV (SEQ ID NO: 4111); ITKGGIT (SEQ ID NO: 4114); and INSGGPT (SEQ ID NO: 4117).

81. The multispecific antigen-binding protein of any one of claims 1-80, wherein the one or more anti- CDH17 antigen-binding domains comprise a CDR1 comprising an amino acid sequence selected from SEQ ID Nos: 2501, 2506, 2511, 2516, 2521, 2528, 2532, 2537, 2542, 2547, 2552, 2557, 2562, 3055- 3292, 4107, 4110, 4113, 4116, 4192-4220; a CDR2 comprising an amino acid sequence selected from SEQ ID Nos: 2502, 2507, 2512, 2517, 2533, 2538, 2543, 2548, 2553, 2558, 2563, 3293-3530, 4108, 4111, 4114, 4117, 4221-4248; and / or a CDR3 comprising an amino acid sequence selected from SEQ520323195900v2Attorney Docket No: 260525.000128ID Nos: 2503, 2508, 2513, 2518, 2522, 2525, 2529, 2534, 2539, 2544, 2549, 2554, 2559, 2564, 3531- 3768, 4109, 4112, 4115, 4118, 4249-4277.

82. The multispecific antigen-binding protein of any one of claims 1-81, wherein the one or more anti- CDH17 antigen-binding domains comprise:i). a CDR1 comprising an amino acid sequence of GFTLSNYN (SEQ ID NO: 2501), a CDR2 comprising an amino acid sequence of ISRGGRT (SEQ ID NO: 2502), and a CDR3 comprising an amino acid sequence of ALCLLRFETCLEYNRAQYPY (SEQ ID NO: 2503);ii). a CDR1 comprising an amino acid sequence of GSPLDYYA (SEQ ID NO: 2506), a CDR2 comprising an amino acid sequence of ISTSGRCT (SEQ ID NO: 2507), and a CDR3 comprising an amino acid sequence of AAVRSGSDWWTTMTQRHYDF (SEQ ID NO: 2508);iii). a CDR1 comprising an amino acid sequence of RLNFSRTT (SEQ ID NO: 2511), a CDR2 comprising an amino acid sequence of SGWARGRT (SEQ ID NO: 2512), and a CDR3 comprising an amino acid sequence of AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513);iv). a CDR1 comprising an amino acid sequence of GRTFSEPI (SEQ ID NO: 2516), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of AATGDSYRGAYDRPAEYDY (SEQ ID NO: 2518);v). a CDR1 comprising an amino acid sequence of GRTFSTPI (SEQ ID NO: 2521), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of AATGNSYRGAYDRPTEYDY (SEQ ID NO: 2522);vi). a CDR1 comprising an amino acid sequence of GRTFSEPI (SEQ ID NO: 2516), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of AATGDLYRGAYDRPAEYDY (SEQ ID NO: 2525);vii). a CDR1 comprising an amino acid sequence of GRTFSSPI (SEQ ID NO: 2528), a CDR2 comprising an amino acid sequence of LISTGGST (SEQ ID NO: 2517), and a CDR3 comprising an amino acid sequence of VATGNTYRGAYDRPAEYDF (SEQ ID NO: 2529);viii). a CDR1 comprising an amino acid sequence of TRTFNMYA (SEQ ID NO: 2532), a CDR2 comprising an amino acid sequence of ISRSGTNT (SEQ ID NO: 2533), and a CDR3 comprising an amino acid sequence of AAQFSLPVDASPLRRYYY (SEQ ID NO: 2534);ix). a CDR1 comprising an amino acid sequence of GRTFSSYL (SEQ ID NO: 2537), a CDR2 comprising an amino acid sequence of ISWNDRST (SEQ ID NO: 2538), and a CDR3 comprising an amino acid sequence of AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);521323195900v2Attorney Docket No: 260525.000128x). a CDR1 comprising an amino acid sequence of GRTDSILN (SEQ ID NO: 2542), a CDR2 comprising an amino acid sequence of ISWFRGET (SEQ ID NO: 2543), and a CDR3 comprising an amino acid sequence of AARNGGYDLNDYAY (SEQ ID NO: 2544);xi). a CDR1 comprising an amino acid sequence of GIRFSSYA (SEQ ID NO: 2547), a CDR2 comprising an amino acid sequence of ITSGYRT (SEQ ID NO: 2548), and a CDR3 comprising an amino acid sequence of NAGGGRLGY (SEQ ID NO: 2549);xii). a CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554);xiii). a CDR1 comprising an amino acid sequence of GIRFSAYA (SEQ ID NO: 2557), a CDR2 comprising an amino acid sequence of MTAGGMT (SEQ ID NO: 2558), and a CDR3 comprising an amino acid sequence of NVGGQLLGY (SEQ ID NO: 2559);xiv). a CDR1 comprising an amino acid sequence of GSRFSSYA (SEQ ID NO: 2562), a CDR2 comprising an amino acid sequence of ITSGGRT (SEQ ID NO: 2563), and a CDR3 comprising an amino acid sequence of NQGGSKGY (SEQ ID NO: 2564);xv). a CDR1 comprising an amino acid sequence of TRTFDMYA (SEQ ID NO: 3137), a CDR2 comprising an amino acid sequence of INRSGANT (SEQ ID NO: 3375), and a CDR3 comprising an amino acid sequence of AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613);xvi). a CDR1 comprising an amino acid sequence of GGTASEYG (SEQ ID NO: 4107), a CDR2 comprising an amino acid sequence of ISTSGGVT (SEQ ID NO: 4108), and a CDR3 comprising an amino acid sequence of AADGLPYGDWFGDQFDV (SEQ ID NO:4109);xvii) a CDR1 comprising an amino acid sequence of EQTMTGFW (SEQ ID NO: 4110), a CDR2 comprising an amino acid sequence of ISASGSRV (SEQ ID NO: 4111), and a CDR3 comprising an amino acid sequence of AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);xviii) a CDR1 comprising an amino acid sequence of GLRFSNYA (SEQ ID NO: 4113), a CDR2 comprising an amino acid sequence of ITKGGIT (SEQ ID NO: 4114), and a CDR3 comprising an amino acid sequence of NMHRSYDISFYDN (SEQ ID NO: 4115); orxix). a CDR1 comprising an amino acid sequence of GGTVSGYA (SEQ ID NO: 4116), a CDR2 comprising an amino acid sequence of INSGGPT (SEQ ID NO: 4117), and a CDR3 comprising an amino acid sequence of RRYDDYGS (SEQ ID NO: 4118).522323195900v2Attorney Docket No: 260525.00012883. The multispecific antigen-binding protein of claim 1-82, wherein the one or more anti-CDH17 antigen-binding domains comprise:i). a CDR1 comprising an amino acid sequence of GRTDSILN (SEQ ID NO: 2542), a CDR2 comprising an amino acid sequence of ISWFRGET (SEQ ID NO: 2543), and a CDR3 comprising an amino acid sequence of AARNGGYDLNDYAY (SEQ ID NO: 2544);ii). a CDR1 comprising an amino acid sequence of GIRFSSYA (SEQ ID NO: 2547), a CDR2 comprising an amino acid sequence of ITSGYRT (SEQ ID NO: 2548), and a CDR3 comprising an amino acid sequence of NAGGGRLGY (SEQ ID NO: 2549);iii). a CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554);iv). a CDR1 comprising an amino acid sequence of RLNFSRTT (SEQ ID NO: 2511), a CDR2 comprising an amino acid sequence of SGWARGRT (SEQ ID NO: 2512), and a CDR3 comprising an amino acid sequence of AARDSRRGGLFADLNEYDY (SEQ ID NO: 2513);v). a CDR1 comprising an amino acid sequence of TRTFDMYA (SEQ ID NO: 3137), a CDR2 comprising an amino acid sequence of INRSGANT (SEQ ID NO: 3375), and a CDR3 comprising an amino acid sequence of AAQFSLPVDATPLRRYAH (SEQ ID NO: 3613);vi). a CDR1 comprising an amino acid sequence of GRTFSSYL (SEQ ID NO: 2537), a CDR2 comprising an amino acid sequence of ISWNDRST (SEQ ID NO: 2538), and a CDR3 comprising an amino acid sequence of AATRKTRRSTVAGTEVDY (SEQ ID NO: 2539);vii). a CDR1 comprising an amino acid sequence of GGTASEYG (SEQ ID NO: 4107), a CDR2 comprising an amino acid sequence of ISTSGGVT (SEQ ID NO: 4108), and a CDR3 comprising an amino acid sequence of AADGLPYGDWFGDQFDV (SEQ ID NO: 4109);viii). a CDR1 comprising an amino acid sequence of EQTMTGFW (SEQ ID NO: 4110), a CDR2 comprising an amino acid sequence of ISASGSRV (SEQ ID NO: 4111), and a CDR3 comprising an amino acid sequence of AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);ix). a CDR1 comprising an amino acid sequence of GLRFSNYA (SEQ ID NO: 4113), a CDR2 comprising an amino acid sequence of ITKGGIT (SEQ ID NO: 4114), and a CDR3 comprising an amino acid sequence of NMHRSYDISFYDN (SEQ ID NO: 4115); orx). a CDR1 comprising an amino acid sequence of GGTVSGYA (SEQ ID NO: 4116), a CDR2 comprising an amino acid sequence of INSGGPT (SEQ ID NO: 4117), and a CDR3 comprising an amino acid sequence of RRYDDYGS (SEQ ID NO: 4118).523323195900v2Attorney Docket No: 260525.00012884. The multispecific antigen-binding protein of claim 1-83, wherein the one or more anti-CDH17 antigen-binding domains comprise:a), a CDR1 comprising an amino acid sequence of GGTASEYG (SEQ ID NO: 4107), a CDR2 comprising an amino acid sequence of ISTSGGVT (SEQ ID NO: 4108), and a CDR3 comprising an amino acid sequence of AADGLPYGDWFGDQFDV (SEQ ID NO: 4109);b). a CDR1 comprising an amino acid sequence of EQTMTGFW (SEQ ID NO: 4110), a CDR2 comprising an amino acid sequence of ISASGSRV (SEQ ID NO: 4111), and a CDR3 comprising an amino acid sequence of AFNKWGRLSADLDDYFR (SEQ ID NO: 4112);c). a CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554); ord). a CDR1 comprising an amino acid sequence of GGTVSGYA (SEQ ID NO: 4116), a CDR2 comprising an amino acid sequence of INSGGPT (SEQ ID NO: 4117), and a CDR3 comprising an amino acid sequence of RRYDDYGS (SEQ ID NO: 4118).

85. The multispecific antigen-binding protein of any one of claims 1-84, wherein the one or more anti- CDH17 antigen-binding domains comprise at least one anti-CDH17 single-domain antibody.

86. The multispecific antigen-binding protein of claim 85, wherein the anti-CDH17 single-domain antibody is an anti-CDH17 VHH, an anti-CDH17 VNAR, or an anti-CDH17 VH domain.

87. The multispecific antigen-binding protein of claim 86, wherein the anti-CDH17 single-domain antibody is an anti-CDH17 VHH.

88. The multispecific antigen-binding protein of claim 87, wherein the anti-CDH17 VHH is a camelid anti- CDH17 VHH.

89. The multispecific antigen-binding protein of claim 88, wherein the camelid anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2504, 2509, 2514, 2519, 2523, 2526, 2530, 2535, 2540, 2545, 2550, 2555, 2560, 2565, 2605-2842, 4119-4122, 4143-4171, and 4278-4306, or an amino acid sequence having at least 75% identity thereto.524323195900v2Attorney Docket No: 260525.00012890. The multispecific antigen-binding protein of claim 89, wherein the camelid anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2545, 2555, 4119, 4120, and 4122, or an amino acid sequence having at least 75% identity thereto.

91. The multispecific antigen-binding protein of claim 87, wherein the anti-CDH17 VHH is a humanized anti-CDH17 VHH.

92. The multispecific antigen-binding protein of claim 91, wherein the humanized anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2505, 2510, 2515, 2520, 2524, 2527, 2531, 2536, 2541, 2546, 2551, 2556, 2561, 2566, 2843-3054, 4123-4126, 4172-4191, 4651-4659, and 4691, or a sequence having at least 75% identity thereto.

93. The multispecific antigen-binding protein of claim 91 or 92, wherein the humanized anti-CDH17 VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 2515, 2541, 2556, 4123, 4124, 4126, 4651-4659, and 4691, or a sequence having at least 75% identity thereto.

94. The multispecific antigen-binding protein of any one of claims 1-93, wherein the one or more anti- CDH17 antigen-binding domains bind to human CDH17.

95. The multispecific antigen-binding protein of claim 94, wherein the one or more anti-CDH17 antigenbinding domains bind to human CDH17 with a KDof about 2.2xl0“7M or less.

96. The multispecific antigen-binding protein of claim 95, wherein the one or more anti-CDH17 antigenbinding domains bind to human CDH17 with a KDof about 1.5xl0“7M or less.

97. The multispecific antigen-binding protein of any one of claims 1-93, wherein the one or more anti- CDH17 antigen-binding domains bind to cyno CDH17.

98. The multispecific antigen-binding protein of claim 97, wherein the one or more anti-CDH17 antigenbinding domains bind to cyno CDH17 with a KDof about 2xl0“8M or less.525323195900v2Attorney Docket No: 260525.00012899. The multispecific antigen-binding protein of any one of claims 1-93, wherein the one or more anti- CDH17 antigen-binding domains bind to mouse CDH17.

100. The multispecific antigen-binding protein of any one of claims 1-99, wherein the one or more anti-CDH17 antigen-binding domains bind to the extracellular cadherin domain 6 (EC6) and / or extracellular cadherin domain 7 (EC7) of human CDH17.

101. The multispecific antigen-binding protein of any one of claims 1-99, wherein the one or more anti-CDH17 antigen-binding domains bind to an extracellular region C-terminal to the tripeptide motif, RGD, in EC6 of human CDH17.

102. The multispecific antigen-binding protein of any one of claims 1-99, wherein the one or more anti-CDH17 antigen-binding domains bind to the extracellular cadherin domain 1 (ECI) of human CDH17.

103. The multispecific antigen-binding protein of any one of claims 1-102, wherein the one or more anti-CDH17 antigen-binding domains bind to the same epitope on CDH17.

104. The multispecific antigen-binding protein of any one of claims 1-102, wherein the one or more anti-CDH17 antigen-binding domains bind to different epitopes on CDH17.

105. The multispecific antigen-binding protein of any one of claims 1-104, whereina) the one or more anti-TRAILR2 antigen-binding domains comprisei). a CDR1 comprising an amino acid sequence of GRSFGDFA (SEQ ID NO: 52), a CDR2 comprising an amino acid sequence of IRWTGDT (SEQ ID NO: 53), and a CDR3 comprising an amino acid sequence of TAGPTMSYSRGGEF (SEQ ID NO: 54);ii). a CDR1 comprising an amino acid sequence of GSNFRILS (SEQ ID NO: 2195), a CDR2 comprising an amino acid sequence of ITSDDTT (SEQ ID NO: 2196), and a CDR3 comprising an amino acid sequence of HSRWYNL (SEQ ID NO: 2197);iii). a CDR1 comprising an amino acid sequence of GSIFGGYN (SEQ ID NO: 2189), a CDR2 comprising an amino acid sequence of IFISGNN (SEQ ID NO: 2190), and a CDR3 comprising an amino acid sequence of RAYNDGGEY (SEQ ID NO: 2191);526323195900v2Attorney Docket No: 260525.000128iv). a CDR1 comprising an amino acid sequence of GRSFGAQGMEG (SEQ ID NO: 72), a CDR2 comprising an amino acid sequence of IKWTGNT (SEQ ID NO: 77), and a CDR3 comprising an amino acid sequence of TAGPSISYSRGGEY (SEQ ID NO: 78); orv). a CDR1 comprising an amino acid sequence of GRTFSSYA (SEQ ID NO: 28), a CDR2 comprising an amino acid sequence of ISWNGDIT (SEQ ID NO: 91), and a CDR3 comprising an amino acid sequence of TAGQSISLSQGGEY (SEQ ID NO: 92);andb) the one or more anti-CDH17 antigen-binding domains comprisea CDR1 comprising an amino acid sequence of GMRFSQYA (SEQ ID NO: 2552), a CDR2 comprising an amino acid sequence of IFKDGTT (SEQ ID NO: 2553), and a CDR3 comprising an amino acid sequence of NAGGAALGY (SEQ ID NO: 2554).

106. The multispecific antigen-binding protein of any one of claims 6-9, 15-16, and 20-24, wherein the antigen binding domain(s) that specifically bind to HSA comprise:i). a CDR1 comprising an amino acid sequence of GFTFTDYS (SEQ ID NO: 4678), a CDR2 comprising an amino acid sequence of TRTTGGSI (SEQ ID NO: 4679), and a CDR3 comprising an amino acid sequence of LAYRQKGSNSPLIV (SEQ ID NO: 4680); orii). a CDR1 comprising an amino acid sequence of GFTFTDYS (SEQ ID NO: 4678), a CDR2 comprising an amino acid sequence of TRTTGGSI (SEQ ID NO: 4679), and a CDR3 comprising an amino acid sequence of LAYRQKGSNSPLII (SEQ ID NO: 4681).

107. The multispecific antigen-binding protein of any one of claims 6-9, 15-16, 20-24 and 106, wherein the one or more anti-HSA antigen-binding domains comprise at least one anti-HSA singledomain antibody.

108. The multispecific antigen-binding protein of claim 107, wherein the anti-HSA single-domain antibody is an anti-HSA VHH, an anti-HSA VNAR, or an anti-HSA VH domain.

109. The multispecific antigen-binding protein of claim 108, wherein the anti-HSA single-domain antibody is an anti-HSA VHH.527323195900v2Attorney Docket No: 260525.000128110. The multispecific antigen-binding protein of any one of claims 106-109, wherein the anti-HSA VHH comprises an amino acid sequence selected from any one of SEQ ID NOs: 4660 and 4661, or an amino acid sequence having at least 75% identity thereto.

111. The multispecific antigen-binding protein of any one of claims 1-110, wherein the antigenbinding protein comprises one or more modifications that reduce binding of said antigen-binding protein by pre-existing antibodies found in human blood or serum.

112. The multispecific antigen-binding protein of claim 111, wherein the one or more antigen-binding domains are one or more VHHs, and wherein the amino acid sequence at the carboxy-terminus of at least one VHH starting from position 111 according to Chothia comprises VAGG (SEQ ID NO: 2405) or VPAG (SEQ ID NO: 2406).

113. The multispecific antigen-binding protein of any one of claims 1-112, wherein the one or more antigen-binding domains are one or more VHHs, and wherein the amino acid residue Glu at the first position of at least one VHH is replaced with Asp (EID).

114. The multispecific antigen-binding protein of any one of claims 7 and 25-113, wherein the immunoglobulin Fc region is an Fc region of a human immunoglobulin.

115. The multispecific antigen-binding protein of claim 114, wherein the immunoglobulin Fc region is an Fc region of human IgGl, lgG2, lgG3 or lgG4, or a variant thereof.

116. The multispecific antigen-binding protein of claim 115, wherein the immunoglobulin Fc region is an Fc region of human IgGl, or a variant thereof.

117. The multispecific antigen-binding protein of claim 116, wherein the Fc region of human IgGl comprises one or more mutations selected from Leu234Ala (L234A), Leu234Gly (L234G), Leu234Ser (L234S), Leu234Thr (L234T), Leu235Ala (L235A), Leu235Glu (L235E), Leu235Ser (L235S), Leu235Thr (L235T), Leu235Val (L235V), Leu235Gln (L235Q), Gly236Arg (G236R), Met252Tyr (M252Y), Ser254Thr (S254T), Thr256Glu (T256E), Asp265Asn (D265N), Asp265Ala (D265A), Asp270Asn528323195900v2Attorney Docket No: 260525.000128(D270N), Ser298Asn (S298N), Asn297Ala (N297A), Pro329Ala (P329A), Pro239Gly (P329G), Asn325Glu (N325E) and / or Ala327Ser (A327S) according to EU numbering.

118. The multispecific antigen-binding protein of claim 117, wherein the Fc region of human IgGl comprises a set of mutations selected from1). L234A and L235A;2). L234A, L235A, and P329A;3). D265A, N297A and P329A;4). L234A, L235A, and G237A;5). L234G, L235S, and G236R;6). L234S, L235T, and G236R;7). L234S, L235V, and G236R;8). L234T, L235Q, and G236R;9). L234T, L235T, and G236R;10). L234A, L235A, and P329G; and11). M252Y, S254T, and T256E.

119. The multispecific antigen-binding protein of claim 115, wherein the immunoglobulin Fc region is an Fc region of human lgG4, or a variant thereof.

120. The multispecific antigen-binding protein of claim 119, wherein the Fc region of human lgG4 comprises one or more mutations selected from Ser228Pro (S228P), Leu235Glu (L235E), Leu235Ala (L235A), Phe234Ala (F234A), and / or Pro329Gly (P329G) according to EU numbering.

121. The multispecific antigen-binding protein of claim 120, wherein the Fc region of human lgG4 comprises a set of mutations selected from1). S228P and L235E;2). S228P and L235A;3). S228P, F234A, and L235E;4). S228P, F234A, and L235A; and5) P329G, S228P, and L235E.529323195900v2Attorney Docket No: 260525.000128122. The multispecific antigen-binding protein of any of claims 114-115, wherein the Fc region comprises an amino acid sequence selected from:DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO: 2152);DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG(SEQ ID NO: 2153);ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVWDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL GK (SEQ ID NO: 2091); or ESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL GK (SEQ ID NO: 2156).

123. The multispecific antigen-binding protein of any of claims 1-122, wherein the multispecific antigen-binding protein comprises at least one linker, wherein said linker is selected from the group consisting of SEQ ID NOs: 2088-2090, 2092-2125, 2132-2148, 2180, 2408, 4007-4041, 4048-4067, and 4683.

124. The multispecific antigen-binding protein of claim 123, wherein the at least one linker is a rigid linker.

125. The multispecific antigen-binding protein of claim 124, wherein the at least one rigid linker is selected from the group consisting of GGGGSPAPAPAPAPAPAPAPAPGGGS (SEQ ID NO: 2088);PAPAPAPAPAPAPAPAP (SEQ ID NO: 2132), GGGGSPAPAPAPAPAPAPAPAPGGGGS (SEQ ID NO: 2135), GGGGSPAPAPAPAPGGGGS (SEQ ID NO: 4698), GGGGSPAPAPAPAPAPAPAPAPAPAPAPAPGGGGS (SEQ530323195900v2Attorney Docket No: 260525.000128ID NO: 4699), and A(EAAAK)nA (SEQ ID NO: 2148), where n is any integer, e.g., 1, 2, 3, 4, 5, 6, or 7, 8, 9 or 10.

126. The multispecific antigen-binding protein of claim 123, wherein the at least one linker is a flexible linker.

127. The multispecific antigen-binding protein of claim 126, wherein the at least one flexible linker is selected from the group consisting of GGGGSGGGS (SEQ ID NO: 4683); Gn(SEQ ID NO: 4700, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10); GnS (SEQ ID NO: 2136, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10), SGn(SEQ ID NO: 2137, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, or 7, 8, 9 or 10), and (GGGGS)n(SEQ ID NO: 2138, where n is any integer, e.g., 1, 2, 3, 4, 5, 6, or 7, 8, 9 or 10).

128. The multispecific antigen-binding protein of any of claims 1-3, wherein the multispecific antigenbinding protein comprises the amino acid sequence of any one of SEQ ID NOs: 4500-4506, 4509- 4524, and 4526-4650, or a sequence having at least 70% identity thereto.

129. The multispecific antigen-binding protein of any of claims 1-6 and 8-24, comprising a single polypeptide chain, said single polypeptide chain comprising an amino acid sequence selected from any one of SEQ ID NOs: 4500-4506, 4509-4524, and 4526-4584, or a sequence having at least 70% identity thereto.

130. The multispecific antigen-binding protein of any of claims 1-3 and 25-38, comprising a first polypeptide chain and a second polypeptide chain, each polypeptide chain comprising an amino acid sequence selected from any one of SEQ ID NOs: 4585-4650, or a sequence having at least 70% identity thereto.

131. The multispecific antigen-binding protein of claim 130, wherein each polypeptide chain comprises an amino acid sequence selected from any one of SEQ ID NOs: 4587-4589, 4603, 4625, 4626, 4633, 4641, 4643, and 4644, or a sequence having at least 70% identity thereto.

132. A conjugate comprising the multispecific antigen-binding protein of any one of claims 1-131, wherein the multispecific antigen-binding protein is conjugated to a second moiety.531323195900v2Attorney Docket No: 260525.000128133. The conjugate of claim 132, wherein the second moiety is selected from a detectable label, a drug, a toxin, a radionuclide, an enzyme, an immunomodulatory agent, a cytokine, a cytotoxic agent, a chemotherapeutic agent, and a diagnostic agent, or a combination thereof.

134. A polynucleotide molecule encoding the multispecific antigen-binding protein of any one of claims 1-131.

135. The polynucleotide molecule of claim 134, which comprises the nucleotide sequence of any one of SEQ ID NOs: 57-61, 131-145, 1731-2053, 2071-2087, 2193, 2199, 2205, 2278-2297, 2567-2580, 3769-4006, 4127-4130, and 4307-4335 or a nucleotide sequence having at least 70% identity thereto.

136. The polynucleotide molecule of claim 135, which comprises the nucleotide sequence of any one of SEQ ID NOs: 57-61, 131-145, 1731-2053, 2071-2087, 2193, 2199, 2205, and 2278-2297, or a nucleotide sequence having at least 70% identity thereto.

137. The polynucleotide molecule of claim 135 or 136, which comprises the nucleotide sequence of any one of SEQ ID NOs: 57-61, 131-145, 2071-2087, 2193, 2199, and 2205, or a nucleotide sequence having at least 70% identity thereto.

138. The polynucleotide molecule of any one of claims 135-137, which comprises the nucleotide sequence of any one of SEQ ID NOs: 132, 137, 2193, 2199, and 2205, or a nucleotide sequence having at least 70% identity thereto.

139. The polynucleotide molecule of claim 135, which comprises the nucleotide sequence of any one of SEQ ID NOs: 2567-2580, 3769-4006, 4127-4130, and 4307-4335, or a nucleotide sequence having at least 70% identity thereto.

140. The polynucleotide molecule of claim 139, which comprises the nucleotide sequence of any one of SEQ ID NOs: 2567-2580, 3851, and 4127-4130, or a nucleotide sequence having at least 70% identity thereto.532323195900v2Attorney Docket No: 260525.000128141. A recombinant vector comprising the polynucleotide molecule of any one of claims 134-140.

142. A host cell comprising the polynucleotide molecule of any one of claims 134-140, or the recombinant vector of claim 141.

143. A kit comprising the multispecific antigen-binding protein of any one of claims 1-131, the conjugate of any one of claims 132-133, the polynucleotide molecule of any one of claims 134-140, the recombinant vector of claim 141, or the host cell of claim 142, and, optionally, instructions and / or packaging for the same.

144. A pharmaceutical composition comprising the multispecific antigen-binding protein of any one of claims 1-131, the conjugate of any one of claims 132-133, the polynucleotide molecule of any one of claims 134-140, or the recombinant vector of claim 141, and a pharmaceutically acceptable carrier and / or excipient.

145. A method for preparing a multispecific antigen-binding protein that specifically binds TNF- related apoptosis-inducing ligand receptor 2 (TRAILR2) and cadherin 17 (CDH17), comprising the steps of:a), culturing the host cell of claim 142 in a culture medium under conditions suitable for expression of the multispecific antigen-binding protein; andb). isolating the multispecific antigen-binding protein from the host cell and / or the culture medium.

146. A method for inducing cell death in a cell expressing TNF-related apoptosis-inducing ligand receptor 2 (TRAILR2) and / or cadherin 17 (CDH17), said method comprising contacting the cell with an effective amount of the antigen-binding protein of claim 1-131 or the conjugate of any one of claims 132-133.

147. The method of claim 146, wherein said contacting occurs in vitro.

148. The method of claim 146, wherein said contacting occurs in vivo.533323195900v2Attorney Docket No: 260525.000128149. The method of claim 148, wherein the cell expressing TRAILR2 and / or CDH17 is in a subject in need thereof.

150. The method of claim 149, wherein the method further comprises administering the multispecific antigen-binding protein or the conjugate into the subject.

151. The method of claim any one of claims 146-150, wherein the cell expressing TRAILR2 and / or CDH17 is a cell of a cancer.

152. The method of claim 151, wherein the cancer is a solid tumor.

153. The method of claim 151, wherein the cancer is selected from adenoid cystic carcinoma, adrenal gland tumor, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxia-telangiectasia, Beckwith-Wiedemann syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dube syndrome, bladder cancer, bone cancer (sarcoma of bone), brain stem glioma, brain tumor, breast cancer, inflammatory breast cancer, metastatic breast cancer, male breast cancer, Carney complex, central nervous system tumors (brain and spinal cord), cervical cancer, childhood cancer, colorectal cancer, Cowden syndrome, craniopharyngioma, desmoid tumor, desmoplastic infantile ganglioglioma, childhood tumor, ependymoma, esophageal cancer, Ewing sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial malignant melanoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell cancer, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal carcinoma, HIV / AIDS-related cancer, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumor, laryngeal and hypopharyngeal cancer, Li-Fraumeni syndrome, liver cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, lynch syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia type 1, multiple endocrine neoplasia type 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndromes (MDS), nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the lung, neuroendocrine tumor of the pancreas,534323195900v2Attorney Docket No: 260525.000128neuroendocrine tumors, neurofibromatosis type 1, neurofibromatosis type 2, nevoid basal cell carcinoma syndrome, oral and oropharyngeal cancer, osteosarcoma, ovarian, fallopian tube, and peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers syndrome, pheochromocytoma and paraganglioma, pituitary gland tumor, pleuropulmonary blastoma, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, Kaposi sarcoma, soft tissue sarcomas, skin cancer (non-melanoma), small bowel cancer, stomach cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis complex, uterine cancer, vaginal cancer, Von Hippel-Lindau syndrome, vulvar cancer, Werner syndrome, Wilms tumor, and xeroderma pigmentosum.

154. The method of claim 151, wherein the cancer is selected from colorectal cancer, familial GIST, familial pancreatic cancer, gastrointestinal stromal tumor (GIST), hereditary diffuse gastric cancer, hereditary pancreatitis, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the pancreas, peritoneal cancer, pancreatic cancer, small bowel cancer, and stomach cancer.

155. The method of claim 151, wherein the cancer is gastrointestinal cancer, breast cancer, or lung cancer.

156. The method of claim 155, wherein the cancer is gastrointestinal cancer.

157. The method of claim 156, wherein the gastrointestinal cancer is colorectal cancer, gastric cancer, esophageal cancer, pancreatic cancer, or cholangiocarcinoma cancer.

158. The method of any one of claims 146-157, wherein the method further comprises contacting the cell with one or more additional therapeutic agents.

159. A method of treating or preventing a cancer in a subject in need thereof, said method comprising administering to the subject an effective amount of the multispecific antigen-binding protein of any one of claims 1-131, or the conjugate of any one of claims 132-133.

160. The method of claim 159, wherein the cancer is a solid tumor.535323195900v2Attorney Docket No: 260525.000128161. The method of claim 159, wherein the cancer is selected from adenoid cystic carcinoma, adrenal gland tumor, amyloidosis, anal cancer, appendix cancer, astrocytoma, ataxia-telangiectasia, Beckwith-Wiedemann syndrome, bile duct cancer (cholangiocarcinoma), Birt-Hogg-Dube syndrome, bladder cancer, bone cancer (sarcoma of bone), brain stem glioma, brain tumor, breast cancer, inflammatory breast cancer, metastatic breast cancer, male breast cancer, Carney complex, central nervous system tumors (brain and spinal cord), cervical cancer, childhood cancer, colorectal cancer, Cowden syndrome, craniopharyngioma, desmoid tumor, desmoplastic infantile ganglioglioma, childhood tumor, ependymoma, esophageal cancer, Ewing sarcoma, eye cancer, eyelid cancer, familial adenomatous polyposis, familial GIST, familial malignant melanoma, familial pancreatic cancer, gallbladder cancer, gastrointestinal stromal tumor (GIST), germ cell tumor, gestational trophoblastic disease, head and neck cancer, hereditary breast and ovarian cancer, hereditary diffuse gastric cancer, hereditary leiomyomatosis and renal cell cancer, hereditary mixed polyposis syndrome, hereditary pancreatitis, hereditary papillary renal carcinoma, HIV / AIDS-related cancer, juvenile polyposis syndrome, kidney cancer, lacrimal gland tumor, laryngeal and hypopharyngeal cancer, Li-Fraumeni syndrome, liver cancer, lung cancer, non-small cell lung cancer, small cell lung cancer, lynch syndrome, mastocytosis, medulloblastoma, melanoma, meningioma, mesothelioma, multiple endocrine neoplasia type 1, multiple endocrine neoplasia type 2, multiple myeloma, MUTYH (or MYH)-associated polyposis, myelodysplastic syndromes (MDS), nasal cavity and paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the lung, neuroendocrine tumor of the pancreas, neuroendocrine tumors, neurofibromatosis type 1, neurofibromatosis type 2, nevoid basal cell carcinoma syndrome, oral and oropharyngeal cancer, osteosarcoma, ovarian, fallopian tube, and peritoneal cancer, pancreatic cancer, parathyroid cancer, penile cancer, Peutz-Jeghers syndrome, pheochromocytoma and paraganglioma, pituitary gland tumor, pleuropulmonary blastoma, prostate cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, Kaposi sarcoma, soft tissue sarcomas, skin cancer (non-melanoma), small bowel cancer, stomach cancer, testicular cancer, thymoma and thymic carcinoma, thyroid cancer, tuberous sclerosis complex, uterine cancer, vaginal cancer, Von Hippel-Lindau syndrome, vulvar cancer, Werner syndrome, Wilms tumor, and xeroderma pigmentosum.

162. The method of claim 159, wherein the cancer is selected from colorectal cancer, familial GIST, familial pancreatic cancer, gastrointestinal stromal tumor (GIST), hereditary diffuse gastric cancer,536323195900v2Attorney Docket No: 260525.000128hereditary pancreatitis, neuroendocrine tumor of the gastrointestinal tract, neuroendocrine tumor of the pancreas, peritoneal cancer, pancreatic cancer, small bowel cancer, and stomach cancer.

163. The method of claim 159, wherein the cancer is gastrointestinal cancer, breast cancer, or lung cancer.

164. The method of claim 163, wherein the cancer is gastrointestinal cancer.

165. The method of claim 164, wherein the gastrointestinal cancer is colorectal cancer, gastric cancer, esophageal cancer, pancreatic cancer, or cholangiocarcinoma cancer.

166. The method of any one of claims 159-165, wherein the method further comprises administering one or more additional therapeutic agents into the subject.

167. The method of claim 158 or 166, wherein the one or more additional therapeutic agents are selected from a chemotherapeutic agent, a vascular endothelial growth factor (VEGF) inhibitor, an epidermal growth factor receptor (EGFR) inhibitor, an apoptosis-inducing agent, and an immunotherapeutic agent, or a combination thereof.

168. The method of claim 167, wherein the chemotherapeutic agent is selected from bleomycin, carboplatin, chlorambucil, cisplatin, colchicine, cyclophosphamide, daunorubicin, doxorubicin or liposomal doxorubicin, mitomycin C, actinomycin, diethylstilbestrol, etoposide, 5-fluorouracil, floxuridine, melphalan, methotrexate, mitomycin, 6-mercaptopurine, teniposide, 6-thioguanine, vincristine and vinblastine, leflunomide, tamoxifen, interferon a-2b, glutamic acid, plicamycin, mercaptopurine, 6-thioguanine, carmustine, BCNU, limousine, CCNU, cytosine arabinose, estramustine, hydroxyurea, procarbazine, busulfan, medroxyprogesterone, estramustine phosphate sodium, ethenyl estradiol, estradiol, megestrol acetate, methyltestosterone, diethylstilbestrol diphosphate, chlorotrianisene, testolactone, melphalan, chlorambucil, mechlorethamine, thiourea, bethamethasone sodium phosphate, dicarbazine, asparagine, mitotane, vincristine sulfate, vinblastine sulfate, FOLFOX (folinic acid, 5-fluorouracil and oxaliplatin), and FOLFIRI (folinic acid, 5- fluorouracil, and irinotecan), or a derivative or combination thereof.537323195900v2Attorney Docket No: 260525.000128169. The method of claim 168, wherein chemotherapeutic agent is an active metabolite of irinotecan, SN-38.

170. The method of claim 167, wherein the VEGF inhibitor is selected from bevacizumab, ramucirumab, regorafenib, and ziv-aflibercept, or a combination thereof.

171. The method of claim 167, wherein the EGFR inhibitor is selected from cetuximab and panitumumab, or a combination thereof.

172. The method of claim 167, wherein the apoptosis-inducing agent is selected from a B-cell lymphoma 2 (BCL2) inhibitor, a BCL-extra-large (BCL-XL) inhibitor, and an inhibitor of apoptosis proteins (IAP) inhibitor, or a combination thereof.

173. The method of claim 167, wherein the immunotherapeutic agent is selected from an anti-CTLA4 agent, an anti-PDl agent, an anti-PD-Ll agent, an anti-LAG3 agent, and an anti-TIM3 agent, or a combination thereof.

174. The method of any one of claims 149-173, wherein the subject is a mammal.

175. The method of claim 174, wherein the mammal is human.538323195900v2

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