Anti-CD73 antibodies and methods of use thereof

Abstract

The present disclosure provides antibodies that specifically bind to CD73 (e.g., human CD73) and antagonize CD73 function. Also provided are anti-CD73 antibodies further comprising a TGF [beta] binding moiety or a VEGF binding moiety. The disclosure additionally provides pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for making these antibodies, and methods of treating subjects using these antibodies.

Description

[0001] This application is a divisional application of the patent application filed on March 8, 2019, with a priority date of March 9, 2018, application number 201980018301.8, entitled "Anti-CD73 Antibody and Method of Using Thereof".

[0002] Related applications

[0003] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 640,850, filed March 9, 2018, the entire disclosure of which is incorporated herein by reference. Technical Field

[0004] This disclosure relates to antibodies that specifically bind to CD73 (e.g., human CD73) and methods of using them. background

[0006] CD73, also known as extracellular 5'-nucleotidase, extracellular 5'-NT, 5'-NT, and NT5E, is a glycosylphosphatidylinositol (GPI)-linked cell surface enzyme. The enzymatic activity of CD73 catalyzes the hydrolysis of extracellular adenosine monophosphate (AMP) to adenosine. This activity, along with the activity of CD39 catalyzing the hydrolysis of extracellular adenosine triphosphate (ATP) to AMP, constitutes the major adenosine production system in the extracellular space. Adenosine is a ligand for many G protein-coupled receptors (GPCRs) and is involved in immune escape, angiogenesis, and tumor motility. CD73 is widely expressed in many tissues and has been found to be upregulated in advanced primary and metastatic tumors. Animal model studies have shown that blocking CD73 activity with anti-CD73 antibodies can inhibit tumor growth and prolong survival by promoting anti-tumor adaptive immunity (Forte et al. (2012) J Immunol. 189(5):2226-33).

[0007] Transforming growth factor-β (TGFβ) is a pleiotropic cytokine that is expressed at elevated levels in advanced primary and metastatic tumors and is involved in tumor cell proliferation, invasion, metastasis, and angiogenesis. TGFβ has also been shown to promote the expansion of immunosuppressive immune populations, such as regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). See Wang et al. (2017) Front Immunol. 8:1934.

[0008] Given the significant roles of human CD73 and TGFβ in regulating the immune response to tumors, therapeutic agents designed to antagonize CD73 activity, or both CD73 activity and TGFβ signaling, hold great promise for cancer treatment. Overview

[0010] This disclosure provides antibodies that specifically bind to CD73 (e.g., human CD73) and antagonize CD73 functions (e.g., enzyme activity that promotes the conversion of AMP to adenosine). Pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for preparing these antibodies are also provided, as well as methods for treating subjects using these antibodies. The antibodies disclosed herein are particularly useful for enhancing immune responses to tumor antigens and, therefore, for treating cancers in subject populations.

[0011] Therefore, in one aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, said antibodies comprising: a heavy chain variable region comprising complementarity-determining regions (CDRs) CDRH1, CDRH2, and CDRH3, and a light chain variable region comprising complementarity-determining regions CDRL1, CDRL2, and CDRL3, wherein:

[0012] (a) CDRH1 contains the amino acid sequence X1X2WX3X4 (SEQ ID NO:13), in which

[0013] X1 is S or N;

[0014] X2 is either S or Y;

[0015] X3 is either I or M; and

[0016] X4 is N or H;

[0017] (b) CDRH2 contains the amino acid sequence X1IYPRX2X3DTNYX4X5KFKX6 (SEQ ID NO:14), in which

[0018] X1 is either R or T;

[0019] X2 is N, A, or S;

[0020] X3 is either G or S;

[0021] X4 is N, A, or S;

[0022] X5 is either G or Q; and

[0023] X6 is either D or G;

[0024] (c) CDRH3 contains the amino acid sequence of LLDYSMDY (SEQ ID NO:7);

[0025] (d) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0026] X1 is N or I; and

[0027] X2 is either Y or S;

[0028] (e) CDRL2 contains the amino acid sequence YTSRLHS (SEQ ID NO: 10); and / or

[0029] (f) CDRL3 contains the amino acid sequence QQGNTLPXT (SEQ ID NO:17), wherein:

[0030] X is either L or W.

[0031] In some embodiments, CDRH1 comprises amino acid sequences selected from the group consisting of SEQ ID NO: 1 and 2. In some embodiments, CDRH2 comprises amino acid sequences selected from the group consisting of SEQ ID NO: 3-6. In some embodiments, CDRL1 comprises amino acid sequences selected from the group consisting of SEQ ID NO: 8 and 9. In some embodiments, CDRL3 comprises amino acid sequences selected from the group consisting of SEQ ID NO: 11 and 12.

[0032] In some implementations,

[0033] (a) CDRH1 contains the amino acid sequence of SSWIN (SEQ ID NO:2);

[0034] (b) CDRH2 contains the amino acid sequence RIYPRX1GDTNYX2GKFKD (SEQ ID NO:15), in which

[0035] X1 is N, A, or S; and

[0036] X2 is N, A, or S;

[0037] (c) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0038] X1 is N or I; and

[0039] X2 is Y or S; and / or

[0040] (d)CDRL3 contains the amino acid sequence QQGNTLPLT (SEQ ID NO:12).

[0041] In some embodiments, CDRH2 comprises amino acid sequences selected from the group consisting of SEQ ID NO:4-6. In some embodiments, CDRL1 comprises amino acid sequences selected from the group consisting of SEQ ID NO:8 and 9.

[0042] In some embodiments, CDRH1, CDRH2, and CDRH3 comprise the amino acid sequences shown in SEQ ID NO:2, 4, and 7; 2, 5, and 7; 1, 3, and 7; or 2, 6, and 7, respectively. In some embodiments, CDRL1, CDRL2, and CDRL3 comprise the amino acid sequences shown in SEQ ID NO:8, 10, and 12; 9, 10, and 12; or 8, 10, and 11, respectively. In some embodiments, CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 comprise the amino acid sequences shown in SEQ ID NO: 2, 4, 7, 8, 10, and 12; 2, 5, 7, 8, 10, and 12; 1, 3, 7, 8, 10, and 11; 2, 4, 7, 8, 10, and 11; 2, 4, 7, 9, 10, and 12; 2, 6, 7, 8, 10, and 12; 2, 5, 7, 9, 10, and 12; or 2, 6, 7, 9, 10, and 12. In some embodiments, CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 comprise SEQ ID NO:2, 4, 7, 8, 10 and 12, respectively; or the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 shown in 2, 5, 7, 8, 10 and 12.

[0043] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, said antibodies comprising heavy chain variable regions comprising CDR CDRH1, CDRH2 and CDRH3, and light chain variable regions comprising complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH3 comprises the amino acid sequence LLDYSMDY (SEQ ID NO:7); and / or CDRL3 comprises the amino acid sequence QQGNTLPLT (SEQ ID NO:12) or QQGNTLPWT (SEQ ID NO:11).

[0044] In some embodiments, the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31. In some embodiments, the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:19-21 and 23-30. In some embodiments, the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:19-21 and 23-30. In some embodiments, the antibody comprises a heavy chain variable region containing at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences selected from the group consisting of SEQ ID NO: 23-30. In some embodiments, the heavy chain variable region contains amino acid sequences selected from the group consisting of SEQ ID NO: 23-30. In some embodiments, the heavy chain variable region contains the amino acid sequence of SEQ ID NO: 24 or 27. In some embodiments, the antibody comprises a heavy chain variable region containing an amino acid sequence derived from a human pedigree sequence as shown in SEQ ID NO: 22.

[0045] In some embodiments, the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO:41 or 42. In some embodiments, the antibody comprises a light chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:33-35 and 37-40. In some embodiments, the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO:33-35 and 37-40. In some embodiments, the antibody comprises a light chain variable region containing at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical amino acid sequences selected from the group consisting of SEQ ID NO:37-40. In some embodiments, the light chain variable region contains amino acid sequences selected from the group consisting of SEQ ID NO:37-40. In some embodiments, the light chain variable region contains the amino acid sequence of SEQ ID NO:37. In some embodiments, the antibody comprises a light chain variable region containing an amino acid sequence derived from a human pedigree sequence as shown in SEQ ID NO:36.

[0046] In some embodiments, the heavy chain variable region and the light chain variable region respectively comprise the amino acid sequences shown in SEQ ID NO: 24 and 37; 27 and 37; 19 and 33; 20 and 34; 21 and 35; 23 and 37; 23 and 38; 24 and 38; 25 and 37; 26 and 37; 28 and 37; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 29 and 37; 30 and 37; 23 and 39; or 23 and 40. In some embodiments, the heavy chain variable region and the light chain variable region respectively comprise the amino acid sequences shown in SEQ ID NO: 24 and 37; or 27 and 37.

[0047] In some embodiments, the antibody comprises a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the same amino acid sequence as SEQ ID NO:27, and the light chain variable region comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the same amino acid sequence as SEQ ID NO:37.

[0048] In some embodiments, the antibody comprises a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the same amino acid sequence as SEQ ID NO:85, and the light chain variable region comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the same amino acid sequence as SEQ ID NO:97.

[0049] In some embodiments, the antibody comprises a heavy chain variable region and / or a light chain variable region, wherein the heavy chain variable region comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the same amino acid sequence as SEQ ID NO:115, and the light chain variable region comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the same amino acid sequence as SEQ ID NO:97.

[0050] In some embodiments, the antibody comprises a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In some embodiments, the heavy chain constant region is IgG1. In some embodiments, the amino acid sequence of IgG1 contains the N297A mutation numbered according to the EU numbering system. In some embodiments, the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 50, 49, 46, 45, 55, or 56. In some embodiments, the antibody comprises a heavy chain containing amino acid sequences selected from the group consisting of SEQ ID NO: 63-88.

[0051] In some embodiments, the antibody comprises a light chain constant region selected from the group consisting of human Igκ and Igλ. In some embodiments, the light chain constant region is Igκ. In some embodiments, the light chain constant region comprises the amino acid sequence of SEQ ID NO: 93 or 89. In some embodiments, the antibody comprises a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 92 and 94-98.

[0052] In some embodiments, the antibody comprises a heavy chain and a light chain, the heavy chain and the light chain each comprising an amino acid sequence shown in any row of Table 4, preferably selected from the group consisting of: SEQ ID NO: 79 and 97; 85 and 97; 77 and 97; 80 and 97; 86 and 97; 64 and 92; 70 and 96; or 78 and 97.

[0053] In some embodiments of any of SEQ ID NO:19, 20, and 65-68, X is glutamine. In some embodiments of any of SEQ ID NO:21, 23-30, 63, 64, and 69-88, X is glutamic acid. In some embodiments of any of SEQ ID NO:19-21, 23-30, and 63-88, X is pyroglutamic acid.

[0054] In another aspect, this disclosure provides isolated antibodies that bind to the same human CD73 epitope as the antibody of any of the preceding claims.

[0055] In some embodiments of any of the foregoing aspects, the antibody is a humanized antibody. In some embodiments, the antibody is antagonistic to human CD73. In some embodiments, the antibody deactivates, reduces, or inhibits the activity of human CD73. In some embodiments, the antibody weakens the ability of human CD73 to convert adenosine monophosphate to adenosine. In some embodiments, the antibody is internalized after binding to cells expressing human CD73.

[0056] In some embodiments, the isolated antibody further comprises a TGFβ-binding moiety. In some embodiments, the TGFβ-binding moiety is linked to the heavy chain variable region. In some embodiments, the TGFβ-binding moiety is linked to the heavy chain variable region via a peptide linker. In some embodiments, the isolated antibody further comprises a TGFβ-binding moiety linked to the heavy chain constant region. In some embodiments, the TGFβ-binding moiety is linked to a C-terminal residue of the heavy chain constant region. In some embodiments, an N-terminal residue of the TGFβ-binding moiety is linked to a C-terminal residue of the heavy chain constant region via a peptide linker. In some embodiments, the peptide linker comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 103-108.

[0057] In some embodiments, the TGFβ binding moiety specifically binds to human TGFβ. In some embodiments, the TGFβ binding moiety comprises an extracellular domain of the human TGFβ receptor. In some embodiments, the human TGFβ receptor is selected from the group consisting of human TGFβR1, TGFβR2, and TGFβR3. In some embodiments, the human TGFβ receptor is human TGFβR2. In some embodiments, the TGFβ binding moiety comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of SEQ ID NO: 109-112. In some embodiments, the TGFβ-binding moiety comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of SEQ ID NO: 109-112. In some embodiments, the TGFβ-binding moiety comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 109-112, preferably SEQ ID NO: 111. In some embodiments, the TGFβ-binding moiety comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 109-112, preferably SEQ ID NO: 111.

[0058] In some embodiments, the isolated antibody comprises a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 113-121, preferably SEQ ID NO: 114 or 115. In some embodiments, the isolated antibody comprises a polypeptide comprising an amino acid sequence selected from SEQ ID NO: 113-121 and a polypeptide comprising an amino acid sequence selected from SEQ ID NO: 92 or 97. In some embodiments, the isolated antibody comprises two polypeptides each comprising an amino acid sequence selected from SEQ ID NO: 113-121 and two polypeptides each comprising an amino acid sequence selected from SEQ ID NO: 92 or 97. In some embodiments, the isolated antibody comprises two polypeptides each comprising an amino acid sequence selected from SEQ ID NO: 115 and two polypeptides each comprising an amino acid sequence selected from SEQ ID NO: 97.

[0059] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, said antibodies comprising a TGFβ-binding moiety. In some embodiments, said antibodies are antagonistic to human CD73. In some embodiments, said antibodies deactivate, reduce, or inhibit the activity of human CD73. In some embodiments, said antibodies attenuate the ability of human CD73 to convert adenosine monophosphate to adenosine. In some embodiments, said antibodies are antagonistic to human CD73 when bound to TGFβ.

[0060] In some embodiments, the TGFβ binding moiety specifically binds to human TGFβ. In some embodiments, the TGFβ binding moiety comprises an extracellular domain of the human TGFβ receptor. In some embodiments, the human TGFβ receptor is selected from the group consisting of human TGFβR1, TGFβR2, and TGFβR3. In some embodiments, the human TGFβ receptor is human TGFβR2. In some embodiments, the TGFβ binding moiety comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of SEQ ID NO: 109-112. In some embodiments, the TGFβ-binding moiety comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of SEQ ID NO: 109-112. In some embodiments, the TGFβ-binding moiety comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 109-112, preferably SEQ ID NO: 111. In some embodiments, the TGFβ-binding moiety comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 109-112, preferably SEQ ID NO: 111.

[0061] In some embodiments, the isolated antibody further comprises a VEGF-binding moiety. In some embodiments, the VEGF-binding moiety is linked to the heavy chain variable region. In some embodiments, the VEGF-binding moiety is linked to the heavy chain variable region via a peptide linker. In some embodiments, the isolated antibody further comprises a VEGF-binding moiety linked to the heavy chain constant region. In some embodiments, the VEGF-binding moiety is linked to a C-terminal residue of the heavy chain constant region. In some embodiments, an N-terminal residue of the VEGF-binding moiety is linked to a C-terminal residue of the heavy chain constant region via a peptide linker. In some embodiments, the peptide linker comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 103-108.

[0062] In some embodiments, the VEGF-binding moiety specifically binds to human VEGF. In some embodiments, the VEGF-binding moiety comprises an extracellular domain of a human VEGF receptor. In some embodiments, the human VEGF receptor is selected from the group consisting of human VEGFR1, VEGFR2, and VEGFR3. In some embodiments, the human VEGF receptor is human VEGFR1. In some embodiments, the VEGF-binding moiety comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence shown in SEQ ID NO:122. In some embodiments, the VEGF-binding moiety consists of at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence shown in SEQ ID NO:122. In some embodiments, the VEGF binding portion comprises the sequence shown in SEQ ID NO:122. In some embodiments, the VEGF binding portion consists of the sequence shown in SEQ ID NO:122.

[0063] In some embodiments, the isolated antibody comprises a polypeptide containing an amino acid sequence selected from the group consisting of SEQ ID NO: 123-127. In some embodiments, the antibody comprises a polypeptide containing an amino acid sequence selected from SEQ ID NO: 123-127 and a polypeptide containing an amino acid sequence selected from SEQ ID NO: 92, 128, and 97. In some embodiments, the antibody comprises two polypeptides containing amino acid sequences selected from SEQ ID NO: 123-127 and two polypeptides containing amino acid sequences selected from SEQ ID NO: 92, 128, and 97.

[0064] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, said antibodies comprising a VEGF-binding moiety. In some embodiments, said antibodies are antagonistic to human CD73. In some embodiments, said antibodies deactivate, reduce, or inhibit the activity of human CD73. In some embodiments, said antibodies attenuate the ability of human CD73 to convert adenosine monophosphate to adenosine. In some embodiments, said antibodies are antagonistic to human CD73 when bound to VEGF.

[0065] In some embodiments, the VEGF-binding moiety specifically binds to human VEGF. In some embodiments, the VEGF-binding moiety comprises an extracellular domain of a human VEGF receptor. In some embodiments, the human VEGF receptor is selected from the group consisting of human VEGFR1, VEGFR2, and VEGFR3. In some embodiments, the human VEGF receptor is human VEGFR1. In some embodiments, the VEGF-binding moiety comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence shown in SEQ ID NO:122. In some embodiments, the VEGF-binding moiety consists of at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence shown in SEQ ID NO:122. In some embodiments, the VEGF-binding moiety comprises the amino acid sequence shown in SEQ ID NO:122. In some embodiments, the VEGF-binding moiety consists of the amino acid sequence shown in SEQ ID NO:122.

[0066] In some embodiments of any of the foregoing aspects, the antibody further comprises a conjugated cytotoxic agent, cell growth inhibitor, toxin, radionuclide, or detectable marker.

[0067] In some embodiments of any of the foregoing aspects, the antibody binds to an epitope located in a region of human CD73, the amino acid sequence of which consists of the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0068] In some embodiments, the antibody binds to at least one residue in the amino acid sequence of SEQ ID NO:90 and / or 91.

[0069] In some embodiments, the antibody binds to one or more amino acid residues of human CD73 selected from the group consisting of Y158, Y161, P165, or D168 according to the amino acid sequence number of SEQ ID NO:99.

[0070] In some embodiments, the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:133 is significantly reduced (e.g., at least 4-, 5-, 6-, 7-, 8-, 9-, or 10-fold) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141. In some embodiments, the maximum binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:133 is significantly reduced (e.g., at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%) relative to the maximum binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0071] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:134 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141.

[0072] In some embodiments, the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:134 is significantly reduced relative to the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0073] In some embodiments, the antibody binds to amino acid residue Y158 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0074] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:59 is significantly reduced relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times).

[0075] In some embodiments, the antibody binds to amino acid residue Y161 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0076] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:60 is significantly reduced relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times).

[0077] In some embodiments, the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:60 is significantly reduced relative to the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0078] In some embodiments, the antibody binds to amino acid residue P165 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0079] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:139 is significantly reduced relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0080] In some embodiments, the antibody binds to amino acid residue D168 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0081] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:140 is significantly reduced relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times).

[0082] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:135 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141.

[0083] In some embodiments, the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:136 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141.

[0084] In some embodiments, the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:137 is not significantly reduced (e.g., not more than 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0085] In some embodiments, the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:142 is significantly reduced relative to the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%), and the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:137 is not significantly reduced relative to the maximum binding of the antibody to a protein containing the amino acid sequence of SEQ ID NO:141 (e.g., reduced by no more than 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0086] In some embodiments, the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:138 is not significantly reduced (e.g., not more than 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0087] In some embodiments, the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is significantly reduced relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%), and the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:138 is not significantly reduced relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141 (e.g., reduced by no more than 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0088] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibody binds to an epitope located in a region of human CD73, the amino acid sequence of which consists of the amino acid sequences of SEQ ID NO: 90 and / or 91.

[0089] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibody binds to at least one residue in the amino acid sequence of SEQ ID NO:90 and / or 91.

[0090] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibodies bind to one or more amino acid residues of human CD73 selected from the group consisting of Y158, Y161, P165 or D168 according to the amino acid sequence number of SEQ ID NO:99.

[0091] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:133 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0092] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:133 is substantially reduced (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%) relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0093] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:134 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0094] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:134 is substantially reduced (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%) relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0095] In another respect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibody binds to amino acid residue Y158 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0096] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:59 is substantially reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0097] In another respect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibody binds to amino acid residue Y161 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0098] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:60 is substantially reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0099] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:60 is substantially reduced (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%) relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0100] In another respect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibody binds to amino acid residue P165 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0101] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:139 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0102] In another respect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the antibody binds to amino acid residue D168 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0103] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:140 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0104] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:135 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0105] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:136 is significantly reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of said antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0106] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is substantially reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:137 is not substantially reduced (e.g., not more than 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0107] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is substantially reduced relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%), and wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:137 is not substantially reduced relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141 (e.g., reduced by no more than 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0108] On the other hand, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is substantially reduced (e.g., at least 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:138 is not substantially reduced (e.g., not more than 4, 5, 6, 7, 8, 9, or 10 times) relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0109] In another aspect, this disclosure provides isolated antibodies that specifically bind to human CD73, wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:142 is substantially reduced relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141 (e.g., reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90%), and wherein the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:138 is not substantially reduced relative to the maximum binding of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141 (e.g., reduced by no more than 30%, 40%, 50%, 60%, 70%, 80%, or 90%).

[0110] In another respect, this disclosure provides compounds or molecules comprising a CD73-binding moiety and a TGFβ-binding moiety.

[0111] In another respect, this disclosure provides pharmaceutical compositions comprising the antibodies disclosed herein and pharmaceutically acceptable carriers or excipients.

[0112] In another aspect, this disclosure provides isolated polynucleotides encoding polypeptides of the antibodies disclosed herein. In some embodiments, the polynucleotide comprises cDNA or mRNA. In another aspect, this disclosure provides vectors comprising polynucleotides. In some embodiments, the vector is a plasmid vector or a viral vector. In another aspect, this disclosure provides recombinant host cells comprising polynucleotides or vectors. In another aspect, this disclosure provides lipid nanoparticles comprising polynucleotides or vectors. In another aspect, this disclosure provides a method for generating antibodies, the method comprising culturing host cells under suitable conditions to express the polynucleotides and generate the antibodies. In another aspect, this disclosure provides pharmaceutical compositions comprising antibodies, polynucleotides, vectors or lipid nanoparticles, and pharmaceutically acceptable carriers or excipients.

[0113] In another aspect, this disclosure provides a method for treating cancer in a subject, the method comprising administering to the subject an effective amount of the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition disclosed herein. In another aspect, this disclosure provides a method for inhibiting cancer metastasis in a subject, the method comprising administering to the subject an effective amount of the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition disclosed herein. In some embodiments, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is administered intravenously, intratumorally, subcutaneously, intradermally, intramuscularly, intrabladderly, intracranially, intracavitarily, or intraventricularly. In some embodiments, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is administered intratumorally.

[0114] In some embodiments, the method further includes administering one or more additional therapeutic agents to the subject. In some embodiments, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition, along with the one or more additional therapeutic agents, are administered co-administered according to the same schedule (e.g., at the same time intervals). In some embodiments, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition, along with the one or more additional therapeutic agents, are administered co-administered according to different schedules (e.g., at different time intervals). In some embodiments, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is administered co-administered with one or more additional therapeutic agents comprising an activator or agonist of an fms-associated tyrosine kinase 3 (FLT3; CD135) receptor, a toll-like receptor (TLR), or an interferon gene stimulator (STING) receptor. In some embodiments, the TLR agonist or activator is selected from the group consisting of TLR2 agonists, TLR3 agonists, TLR7 agonists, TLR8 agonists, and TLR9 agonists. In some embodiments, the STING receptor agonist or activator is selected from the group consisting of: ADU-S100 (MIW-815), SB-11285, MK-1454, SR-8291, AdVCA0848, GSK-532, SYN-STING, MSA-1, SR-8291, 5,6-dimethylxanthenone-4-acetic acid (DMXAA), cyclic GAMP (cGAMP), and cyclic diAMP.In some embodiments, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is co-administered with one or more additional therapeutic agents comprising inhibitors or antagonists of: non-receptor protein tyrosine phosphatase 11 (PTPN11 or SHP2), myeloid leukemia sequence 1 (MCL1) apoptosis regulator; mitogen-activated protein kinase kinase 1 (MAP4K1) (also known as hematopoietic progenitor cell kinase 1 (HPK1)); diacylglycerol kinase alpha (DGKA, DAGK, DAGK1, or DGK-alpha); extracellular 5'-nucleotidase (NT5E or CD73); transforming growth factor beta 1 (TGFB1 or TGFβ); heme oxygenase 1 (HMOX1, HO-1, or HO1); vascular endothelial growth factor A (VEGFA or VEGF); erb-b2 receptor tyrosine kinase 2 (ERBB2). HER2, HER2 / neu, or CD340; epidermal growth factor receptor (EGFR, ERBB, ERBB1, or HER1); ALK receptor tyrosine kinase (ALK, CD246); poly(ADP-ribose) polymerase 1 (PARP1 or PARP); cyclin-dependent kinase 4 (CDK4); cyclin-dependent kinase 6 (CDK6); CC motif chemokine receptor 8 (CCR8, CDw198); CD274 molecule (CD274, PDL1, or PD-L1); programmed cell death 1 (PDCD1, PD1, or PD-1); and / or cytotoxic T-lymphocyte-associated protein 4 (CTLA4, CTLA-4, CD152). In some embodiments, the inhibitor comprises an antigen-binding molecule, an antibody, or an antigen-binding fragment thereof. In some embodiments, the inhibitor comprises a small organic molecule. In some embodiments, the inhibitor of MCL1 is selected from the group consisting of: AMG-176, AMG-397, S-64315, AZD-5991, 483-LM, A1210477, UMI-77, and JKY-5-037. In some embodiments, the inhibitor of PTPN11 or SHP2 is selected from the group consisting of: TNO155 (SHP-099), RMC-4550, JAB-3068, and RMC-4630.

[0115] In some embodiments, the one or more additional therapeutic agents are chemotherapeutic agents, antibiological agents, radiotherapy agents, or checkpoint targeting agents. In some embodiments, the one or more antibiological agents or chemotherapeutic agents are selected from the group consisting of: nucleoside analogs (e.g., 5-fluorouracil, gemcitabine, cytarabine), taxanes (e.g., paclitaxel, nab-paclitaxel, docetaxel, cabazitaxel), platinum complexes (cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin).tetranitrate, phenanthriplatin, picoplatin, satraplatin, dicycloplatin, eptaplatin, lobaplatin, miriplatin, dihydrofolate reductase (DHFR) inhibitors (e.g., methotrexate, trimetrexate, pemetrexed), topoisomerase inhibitors (e.g., doxorubicin, daunorubicin, dactinomycin, eniposide, epirubicin, etoposide, idarubicin, irinotecan, mitoxantrone, pixantrone, sobuzoxane, topotecan otecan), irinotecan, MM-398 (irinotecan liposome), vosaroxin and GPX-150, aldoxorubicin, AR-67, growlertinib, AST-2818, avitinib (ACEA-0010), irofulven (MGI-114), alkylating agents (e.g., nitrogen mustard, cyclophosphamide, chlormethine, urethane) Uramustine or uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, temozolomide, carmustine, nitrosoureas (e.g., carmustine, lomustine, streptozocin), alkyl sulfonates (e.g., busulfan), and mixtures thereof.

[0116] In some embodiments, the additional therapeutic agent is a checkpoint target. In some embodiments, the checkpoint target is selected from the group consisting of: antagonist anti-PD-1 antibodies, antagonist anti-PD-L1 antibodies, antagonist anti-PD-L2 antibodies, antagonist anti-CTLA-4 antibodies, antagonist anti-TIM-3 antibodies, antagonist anti-LAG-3 antibodies, antagonist anti-CEACAM1 antibodies, agonist anti-GITR antibodies, antagonist anti-TIGIT antibodies, antagonist anti-VISTA antibodies, agonist anti-CD137 antibodies, and agonist anti-OX40 antibodies. In some embodiments, the additional therapeutic agent is an anti-PD-1 antibody, optionally wherein the anti-PD-1 antibody is pembrolizumab or nivolumab. In some embodiments, the additional therapeutic agent is an inhibitor of indoleamine-2,3-dioxygenase (IDO). In some embodiments, the inhibitor is selected from the group consisting of epacadostat, F001287, indoximod, and NLG919. In some embodiments, the inhibitor is epacadostat. In some embodiments, the additional therapeutic agent is a vaccine. In some embodiments, the vaccine comprises a heat shock protein peptide complex (HSPPC), wherein the HSPPC comprises a heat shock protein complexed with an antigenic peptide. In some embodiments, the heat shock protein is hsc70 and complexed with a tumor-associated antigenic peptide. In some embodiments, the heat shock protein is gp96 and complexed with a tumor-associated antigenic peptide, wherein the HSPPC is derived from a tumor obtained from the subject. In some embodiments, the additional therapeutic agent is etoposide or doxorubicin.

[0117] In another aspect, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is administered in combination with the FOLFOX, FOLFOXIRI, or FOLFIRINOX regimens. In yet another aspect, the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is administered in combination with immunotherapy, immunostimulation therapy, cell therapy, or gene therapy.

[0118] On the other hand, this disclosure provides isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions disclosed herein for the treatment of cancer.

[0119] On the other hand, this disclosure provides the use of the isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions disclosed herein in the preparation of medicaments for the treatment of cancer.

[0120] In certain embodiments of any of the aspects of methods for treating cancer in subjects, this disclosure provides isolated antibodies, polynucleotides, carriers, lipid nanoparticles, or pharmaceutical compositions for the treatment of cancer, or provides use of isolated antibodies, polynucleotides, carriers, lipid nanoparticles, or pharmaceutical compositions in the preparation of a medicament for the treatment of cancer, said cancer being selected from the group consisting of solid tumors, hematologic malignancies, and metastatic lesions. In some embodiments, the cancer is a solid tumor. In some embodiments, the solid tumor is selected from the group consisting of sarcomas, fibroblastic sarcomas, carcinomas, and adenocarcinomas. In some embodiments, the cancer is a hematologic malignancies. In some embodiments, the hematologic malignancies are selected from the group consisting of leukemia, lymphomas, and myelomas. In some embodiments, the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), and chronic myelomonocytic leukemia (CM). ML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma. In some embodiments, the cancer is a solid tumor in or originating from a tissue or organ selected from the group consisting of: bone (e.g., ameloblastoma, aneurysmal bone cyst, angiosarcoma, chondroblastoma, chondroma, chondromycinoma, chondrosarcoma, chordoma, dedifferentiated chondrosarcoma, enchondroma, epithelioid hemangioendothelioma, fibrous osteodystrophy, giant cell tumor of bone, hemangioma and related lesions, osteoblastoma, osteochondroma, osteosarcoma, osteoid osteoma, osteoma, periosteal chondroma, desmoidoma, Ewing sarcoma); lips and oral cavity (e.g., odontogenic ameloblastoma, oral leukoplakia, oral squamous cell carcinoma, primary oral mucosal melanoma); salivary glands (e.g., pleomorphic salivary gland adenoma, salivary gland adenoid cystic carcinoma, salivary gland mucoepidermoid carcinoma, salivary gland Warthin tumor); esophagus (e.g., Barrett's esophagus, dysplasia, and adenocarcinoma);The gastrointestinal tract, including the stomach (e.g., gastric adenocarcinoma, primary gastric lymphoma, gastrointestinal stromal tumor (GIST), metastatic deposits, gastric carcinoid, gastric sarcoma, neuroendocrine carcinoma, primary squamous cell carcinoma of the stomach, gastric adenoacanthoma), intestine and smooth muscle (e.g., intravenous leiomyomatosis), colon (e.g., colorectal adenocarcinoma), rectum, anus; pancreas (e.g., serous neoplasms, including microcystic or macrocystic serous cystadenomas, solid serous cystadenomas, Von Hippel-Landau (VHL) related serous cystic neoplasms, serous cystadenocarcinoma, mucinous cystic neoplasms (MCN), intraductal papillary mucinous neoplasms (IPMN), intraductal eosinophilic papillary neoplasms (IOPN), intraductal tubular neoplasms, cystic acinar neoplasms including acinar cell cystadenoma, acinar cell cystadenocarcinoma, pancreatic adenocarcinoma, invasive pancreatic ductal adenocarcinoma including tubular adenocarcinoma, adenosquamous carcinoma, colloid carcinoma, medullary carcinoma, hepatoid carcinoma, signet ring cell carcinoma, undifferentiated carcinoma, undifferentiated carcinoma with osteoclast-like giant cells, acinar cell carcinoma, neuronal carcinoma. Neuroendocrine neoplasms, neuroendocrine microadenomas, neuroendocrine tumors (NETs), neuroendocrine carcinomas (NECs), including small cell or large cell NECs, insulinomas, gastrinomas, glucagonomas, serotonin-producing NETs, ​​somatostatinomas, angiotensin-releasing vasomotor tumors, solid pseudopapillary neoplasms (SPNs), pancreatoblastomas); gallbladder (e.g., gallbladder cancer and extrahepatic bile duct cancer, intrahepatic bile duct cancer); neuroendocrine (e.g., adrenocortical carcinoma, carcinoid tumors, pheochromocytomas, pituitary adenomas); thyroid (e.g., anaplastic (undifferentiated) carcinoma, medullary carcinoma, eosinophilic tumors, papillary carcinomas, adenomas). Cancer); Liver (e.g., adenoma, combined hepatocellular and cholangiocarcinoma, fibrolamellar carcinoma, hepatoblastoma, hepatocellular carcinoma, mesenchymal, nested stromal epithelial tumors, undifferentiated carcinoma, hepatocellular carcinoma, intrahepatic cholangiocarcinoma, bile duct cystadenocarcinoma, epithelioid hemangioendothelioma, angiosarcoma, embryonal sarcoma, rhabdomyosarcoma, solitary fibroma, teratoma, yolk sac tumor, carcinosarcoma, rhabdomyosarcoma); Kidney (e.g., ALK rearranged renal cell carcinoma, chromophobe renal cell carcinoma, clear cell renal cell carcinoma, clear cell sarcoma, metanephrotic adenoma, metanephrotic fibroma, mucinous tubular and spindle cell carcinoma, nephroma, nephroblastoma (Wilms tumor)). (e.g., papillary adenoma, papillary renal cell carcinoma, renal eosinophilic carcinoma, renal cell carcinoma, succinate dehydrogenase deficient renal cell carcinoma, collecting duct carcinoma); breast (e.g., invasive ductal carcinoma, including but not limited to acinic cell carcinoma, adenoid cystic carcinoma, apocrine carcinoma, cribriform carcinoma, glycogen-rich / clear cell carcinoma, inflammatory carcinoma, lipid-rich carcinoma, medullary carcinoma, metaplastic carcinoma, micropapillary carcinoma, mucinous carcinoma, neuroendocrine carcinoma, eosinophilic carcinoma, papillary carcinoma, sebaceous gland carcinoma, secretory breast cancer, tubular carcinoma, lobular carcinoma, including but not limited to pleomorphic carcinoma, signet ring cell carcinoma, peritoneum (e.g., mesothelioma, primary peritoneal carcinoma);Female sex organ tissues, including the ovaries (e.g., choriocarcinoma, epithelial tumors, germ cell tumors, sex cord-stromal tumors), fallopian tubes (e.g., serous adenocarcinoma, mucinous adenocarcinoma, endometrioid adenocarcinoma, clear cell adenocarcinoma, transitional cell carcinoma, squamous cell carcinoma, undifferentiated carcinoma, Müllerian tumor, adenosarcoma, leiomyosarcoma, teratoma, germ cell tumors, choriocarcinoma, trophoblastic tumors), uterus (e.g., cervical cancer, endometrial polyps, endometrial hyperplasia, epithelial intraepithelial carcinoma (EIC), endometrial cancer (e.g., endometrioid carcinoma, serous adenocarcinoma, mucinous adenocarcinoma, endometrioid adenocarcinoma, clear cell adenocarcinoma, transitional cell carcinoma, squamous cell carcinoma, undifferentiated carcinoma, Müllerian tumor, adenosarcoma, leiomyosarcoma, teratoma, germ cell tumor, choriocarcinoma, trophoblastic tumor), and endometrial cancer (e.g., endometrioid carcinoma, serous adenocarcinoma, endometrioid carcinoma, choriocarcinoma, trophoblastic tumor). Hypoxetine carcinoma, clear cell carcinoma, mucinous carcinoma, squamous cell carcinoma, transitional carcinoma, small cell carcinoma, undifferentiated carcinoma, mesenchymal neoplasm formation), leiomyomas (e.g., endometrial stromal nodules, leiomyosarcoma, endometrial stromal sarcoma (ESS), mesenchymal tumors), mixed epithelial and mesenchymal tumors (e.g., adenofibroma, carcinoma fibroma, adenosarcoma, carcinosarcoma (malignant mixed mesodermal sarcoma - MMMT), endometrial stromal tumors, malignant mixed Müllerian tumors of the endometrium, gestational trophoblastic tumors (partial hydatiform mole) Mole, complete hydatidiform mole, invasive hydatidiform mole, placental site tumors); vulva, vagina; male sex organs, including the prostate, testes (e.g., germ cell tumors, seminoma), penis; bladder (e.g., squamous cell carcinoma, urothelial carcinoma, bladder urothelial carcinoma); brain (e.g., gliomas (e.g., astrocytomas, including non-invasive, low-grade, anaplastic, glioblastomas; oligodendrogliomas, ependymomas), meningiomas, gangliogliomas, Schwann cell tumors (schwannomas), craniopharyngiomas, chordomas, non-Hodgkin lymphomas, pituitary tumors); eye (e.g., retinoma, retinoblastoma, ocular melanoma, retrouvian melanoma, iris hamartoma); head and neck (e.g., Nasopharyngeal carcinoma, endolymphatic cystic tumor (ELST), epidermoid carcinoma, laryngeal carcinoma including squamous cell carcinoma (SCC) (e.g., glottic carcinoma, supraglottic carcinoma, subglottic carcinoma, transglottic carcinoma), carcinoma in situ, verrucous, spindle cell and basaloid SCC, undifferentiated carcinoma, laryngeal adenocarcinoma, adenoid cystic carcinoma, neuroendocrine carcinoma, laryngeal sarcoma), head and neck paraganglioma (e.g., carotid body, tympanic cavity, vagus nerve); thymus (e.g., thymoma); heart (e.g., cardiac myxoma); lung (e.g., small cell carcinoma (SCLC), non-small cell lung cancer (NSCLC), including squamous cell carcinoma (SCC), adenocarcinoma and large cell carcinoma, carcinoid (typical or atypical), carcinosarcoma, pulmonary blastoma, giant cell carcinoma, spindle cell carcinoma, pleural pulmonary blastoma);Lymphomas (e.g., lymphomas, including Hodgkin's lymphoma, non-Hodgkin's lymphoma), Epstein-Barr virus (EBV)-associated lymphoproliferative disorders, including B-cell lymphomas and T-cell lymphomas (e.g., Burkitt lymphoma, large B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, indolent B-cell lymphoma, low-grade B-cell lymphoma, fibrin-associated diffuse large cell lymphoma; primary exudative lymphoma; plasmablastic lymphoma; extranodal NK / T-cell lymphoma (nasal type); peripheral T-cell lymphoma, cutaneous T-cell lymphoma, angioimmunoblastic T-cell lymphoma; follicular lymphoma). T-cell lymphoma; systemic T-cell lymphoma, lymphangioleiomyomatosis; central nervous system (CNS) (e.g., gliomas, including astrocytic tumors such as pilocytic astrocytoma, pilocyxoid astrocytoma, subependymal giant cell astrocytoma, pleomorphic xanthoastrocytoma, diffuse astrocytoma, fibrous astrocytoma, and gemistocytic astrocytoma). astrocytoma, protoplasmic astrocytoma, anaplastic astrocytoma, glioblastoma (e.g., giant cell glioblastoma, glioma, glioblastoma multiforme, and gliomatosis), oligodendroglioma (e.g., oligodendroglioma, anaplastic oligodendroglioma), oligodendroastrocytic tumor (e.g., oligodendroglioma, anaplastic oligodendroglioma), ependymal tumor (e.g., subependymoma, myxopapillary ependymoma, ependymoma (e.g., cellular, papillary, clear cell, elongated cell), anaplastic ependymoma), optic glioma and non-glial tumor (e.g., choroid plexus tumors, neuronal and mixed neuronal glial tumors, pineal region tumors, embryonal tumors). Tumors, including medulloblastomas, meningeal tumors, primary CNS lymphomas, germ cell tumors, pituitary adenomas, cranial nerve and paravertebral nerve tumors, stellate region tumors, neurofibromas, meningiomas, peripheral nerve sheath tumors, peripheral neuroblastomas (including but not limited to neuroblastomas, ganglioneuromas, ganglioneuromas), trisomy 19 ependymomas; neuroendocrine tissues (e.g., paraganglionic systems, including adrenal medulla (pheochromocytoma) and extra-adrenal paragangliomas (extra-adrenal paragangliomas); skin (e.g., clear cell hidradenoma, benign fibrous histiocytoma of the skin, cylindrica, hidradenoma, melanoma (including cutaneous melanoma, mucosal melanoma), pilomatoma, Spitz tumors);and soft tissue (e.g., invasive angiomyxoma, alveolar rhabdomyosarcoma, alveolar soft sarcoma, angiofibroma, hemangioma-like fibrous histiocytoma, synovial sarcoma, biphasic synovial sarcoma, clear cell sarcoma, dermatofibrosarcoma protuberans, desmoid fibromatosis, small round cell tumor, pro-connective tissue proliferative small round cell tumor, elastic fibroma, embryonal rhabdomyosarcoma, Ewing's tumor / primary neuroectodermal tumor (PNET), extraskeletal myxoid chondrosarcoma, extraskeletal osteosarcoma, paravertebral sarcoma, inflammatory myofibroblastic tumor, lipoblastoma, lipoma, chondroid lipoma, liposarcoma / malignant lipomatous tumor, liposarcoma, myxoma). Liposarcoma, fibromyxoid sarcoma, lymphangioleiomyoma, malignant myoepithelioma, soft malignant melanoma, myoepithelial carcinoma, myoepithelioma, myxoinflammatory fibroblastic sarcoma, undifferentiated sarcoma, peridermoma, rhabdomyosarcoma, non-rhabdomyosarcoma soft tissue sarcoma, soft tissue leiomyosarcoma, undifferentiated sarcoma, well-differentiated liposarcoma. In some embodiments, the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal carcinoma, and endometrial cancer.

[0121] On the other hand, this disclosure provides for the use of isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions as disclosed herein in any of the foregoing methods for treating cancer.

[0122] On the other hand, this disclosure provides isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions as disclosed herein for use in any of the foregoing methods for treating cancer.

[0123] The following describes other or alternative embodiments of the invention disclosed herein.

[0124] Implementation Scheme 1. An antibody that binds to human CD73, said antibody comprising: a heavy chain variable region containing complementarity-determining regions (CDRs) CDRH1, CDRH2, and CDRH3, and a light chain variable region containing complementarity-determining regions CDRL1, CDRL2, and CDRL3, wherein:

[0125] (a) CDRH1 contains the amino acid sequence X1X2WX3X4 (SEQ ID NO:13), in which

[0126] X1 is S or N;

[0127] X2 is either S or Y;

[0128] X3 is either I or M; and

[0129] X4 is N or H;

[0130] (b) CDRH2 contains the amino acid sequence X1IYPRX2X3DTNYX4X5KFKX6 (SEQ ID NO:14), in which

[0131] X1 is either R or T;

[0132] X2 is N, A, or S;

[0133] X3 is either G or S;

[0134] X4 is N, A, or S;

[0135] X5 is either G or Q; and

[0136] X6 is either D or G;

[0137] (c) CDRH3 contains the amino acid sequence of LLDYSMDY (SEQ ID NO:7);

[0138] (d) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0139] X1 is N or I; and

[0140] X2 is either Y or S;

[0141] (e) CDRL2 contains the amino acid sequence YTSRLHS (SEQ ID NO: 10); and / or

[0142] (f) CDRL3 contains the amino acid sequence QQGNTLPXT (SEQ ID NO:17), wherein:

[0143] X is either L or W.

[0144] Implementation Scheme 2. The antibody of Implementation Scheme 1, wherein CDRH1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1 and 2.

[0145] Implementation Scheme 3. The antibody of Implementation Scheme 1 or 2, wherein CDRH2 comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 3-6.

[0146] Implementation Scheme 4. The antibody of any of the foregoing implementation schemes, wherein CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 8 and 9.

[0147] Implementation Scheme 5. The antibody of any of the foregoing implementation schemes, wherein CDRL3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 11 and 12.

[0148] Implementation Plan 6. The antibody of any one of the aforementioned implementation plans, wherein:

[0149] (a) CDRH1 contains the amino acid sequence of SSWIN (SEQ ID NO:2);

[0150] (b) CDRH2 contains the amino acid sequence RIYPRX1GDTNYX2GKFKD (SEQ ID NO:15), in which

[0151] X1 is N, A, or S; and

[0152] X2 is N, A, or S;

[0153] (c) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0154] X1 is N or I; and

[0155] X2 is Y or S; and / or

[0156] (d)CDRL3 contains the amino acid sequence QQGNTLPLT (SEQ ID NO:12).

[0157] Implementation Scheme 7. The antibody of Implementation Scheme 6, wherein CDRH2 comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 4-6.

[0158] Implementation scheme 8. The antibody of implementation scheme 6 or 7, wherein CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 8 and 9.

[0159] Implementation Scheme 9. An antibody of any of the foregoing implementation schemes, wherein CDRH1, CDRH2 and CDRH3 respectively contain the amino acid sequences of CDRH1, CDRH2 and CDRH3 shown in SEQ ID NO:2, 4 and 7; 2, 5 and 7; 1, 3 and 7; or 2, 6 and 7.

[0160] Implementation Scheme 10. An antibody of any of the preceding implementation schemes, wherein CDRL1, CDRL2 and CDRL3 respectively comprise the amino acid sequences of CDRL1, CDRL2 and CDRL3 shown in SEQ ID NO: 8, 10 and 12; 9, 10 and 12; or 8, 10 and 11.

[0161] Implementation Scheme 11. The antibody of any of the preceding implementation schemes, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 respectively comprise SEQ ID NO: 2, 4, 7, 8, 10 and 12; 2, 5, 7, 8, 10 and 12; 1, 3, 7, 8, 10 and 11; 2, 4, 7, 8, 10 and 11; 2, 4, 7, 9, 10 and 12; 2, 6, 7, 8, 10 and 12; 2, 5, 7, 9, 10 and 12; or the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 shown in 2, 6, 7, 9, 10 and 12.

[0162] Implementation Scheme 12. The antibody of Implementation Scheme 11, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 respectively contain SEQ ID NO: 2, 4, 7, 8, 10 and 12; or the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 shown in 2, 5, 7, 8, 10 and 12.

[0163] Implementation Scheme 13. An antibody that binds to human CD73, said antibody comprising a heavy chain variable region containing CDR CDRH1, CDRH2 and CDRH3, and a light chain variable region containing complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH3 contains the amino acid sequence LLDYSMDY (SEQ ID NO:7); and / or CDRL3 contains the amino acid sequence QQGNTLPLT (SEQ ID NO:12) or QQGNTLPWT (SEQ ID NO:11).

[0164] Implementation Scheme 14. An antibody according to any of the preceding implementation schemes, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31.

[0165] Implementation Scheme 15. An antibody according to any of the preceding embodiments, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 19-21 and 23-30.

[0166] Implementation Scheme 16. The antibody of Implementation Scheme 15, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 19-21 and 23-30.

[0167] Implementation Scheme 17. An antibody of any of the preceding embodiments, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of: SEQ ID NO:23-30.

[0168] Implementation Scheme 18. The antibody of Implementation Scheme 17, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:23-30.

[0169] Implementation Scheme 19. The antibody of Implementation Scheme 18, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 24 or 27.

[0170] Implementation Scheme 20. An antibody according to any of the preceding implementation schemes, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence derived from the human lineage sequence shown in SEQ ID NO:22.

[0171] Implementation Scheme 21. An antibody according to any of the preceding implementation schemes, wherein the antibody comprises a light chain variable region comprising an amino acid sequence of SEQ ID NO: 41 or 42.

[0172] Implementation Scheme 22. An antibody of any of the preceding embodiments, wherein the antibody comprises a light chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 33-35 and 37-40.

[0173] Implementation Scheme 23. The antibody of Implementation Scheme 22, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 33-35 and 37-40.

[0174] Implementation Scheme 24. An antibody of any of the preceding embodiments, wherein the antibody comprises a light chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of: SEQ ID NO:37-40.

[0175] Implementation Scheme 25. The antibody of Implementation Scheme 24, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:37-40.

[0176] Implementation Scheme 26. The antibody of Implementation Scheme 25, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:37.

[0177] Implementation Scheme 27. An antibody of any of the preceding implementation schemes, wherein the antibody comprises a light chain variable region comprising an amino acid sequence derived from the human lineage sequence shown in SEQ ID NO:36.

[0178] Implementation Scheme 28. An antibody of any of the preceding implementation schemes, wherein the heavy chain variable region and the light chain variable region respectively comprise the amino acid sequences shown in SEQ ID NO: 24 and 37; 27 and 37; 19 and 33; 20 and 34; 21 and 35; 23 and 37; 23 and 38; 24 and 38; 25 and 37; 26 and 37; 28 and 37; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 29 and 37; 30 and 37; 23 and 39; or 23 and 40.

[0179] Implementation Scheme 29. The antibody of Implementation Scheme 28, wherein the heavy chain variable region and the light chain variable region comprise the amino acid sequences shown in SEQ ID NO: 24 and 37, respectively; or 27 and 37.

[0180] Implementation Scheme 30. An antibody of any of the preceding implementation schemes, wherein the antibody comprises a heavy chain constant region selected from the group consisting of: human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0181] Implementation Scheme 31. The antibody of Implementation Scheme 30, wherein the heavy chain constant region is IgG1.

[0182] Implementation scheme 32. The antibody of implementation scheme 31, wherein the amino acid sequence of IgG1 contains the N297A mutation numbered according to the EU numbering system.

[0183] Implementation Scheme 33. The antibody of Implementation Scheme 32, wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 50, 49, 46, 45, 55 or 56.

[0184] Implementation Scheme 34. An antibody of any of the foregoing implementation schemes, wherein the antibody comprises a heavy chain, the heavy chain comprising an amino acid sequence selected from the group consisting of: SEQ ID NO:63-88.

[0185] Implementation Scheme 35. An antibody of any of the foregoing implementation schemes, wherein the antibody comprises a light chain constant region selected from the group consisting of human Igκ and Igλ.

[0186] Implementation scheme 36. The antibody of implementation scheme 35, wherein the light chain constant region is Igκ.

[0187] Implementation Scheme 37. The antibody of Implementation Scheme 36, wherein the light chain constant region comprises the amino acid sequence of SEQ ID NO: 93 or 89.

[0188] Implementation Scheme 38. An antibody of any of the preceding implementation schemes, wherein the antibody comprises a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 92 and 94-98.

[0189] Implementation Scheme 39. An antibody of any of the foregoing implementation schemes, wherein the antibody comprises a heavy chain and a light chain, the heavy chain and the light chain each comprising an amino acid sequence shown in any row of Table 4, preferably selected from the group consisting of: SEQ ID NO: 79 and 97; 85 and 97; 77 and 97; 80 and 97; 86 and 97; 64 and 92; 70 and 96; or 78 and 97.

[0190] Implementation Scheme 40. An antibody of any one of Implementation Schemes 15-39, wherein X is glutamine in any one of SEQ ID NO: 19, 20 and 65-68.

[0191] Implementation Scheme 41. An antibody of any one of Implementation Schemes 15-39, wherein X is glutamic acid in any one of SEQ ID NO: 21, 23-30, 63, 64, 69-88.

[0192] Implementation Scheme 42. An antibody of any one of Implementation Schemes 15-39, wherein X is pyroglutamic acid in any one of SEQ ID NO: 19-21, 23-30 and 63-88.

[0193] Implementation Scheme 43. An antibody that binds to the same human CD73 epitope as the antibody in any of the preceding implementation schemes.

[0194] Implementation Scheme 44. The antibody of any of the foregoing implementation schemes, wherein the antibody is a humanized antibody.

[0195] Implementation Scheme 45. An antibody according to any of the foregoing implementation schemes, wherein the antibody is antagonistic to human CD73.

[0196] Implementation Scheme 46. The antibody of Implementation Scheme 45, wherein the antibody deactivates, reduces or inhibits the activity of human CD73.

[0197] Implementation Scheme 47. The antibody of Implementation Scheme 45 or 46, wherein the antibody weakens the ability of human CD73 to convert adenosine monophosphate into adenosine.

[0198] Implementation Scheme 48. The antibody of any of the preceding implementation schemes, wherein the antibody is internalized after binding to cells expressing human CD73.

[0199] Implementation Scheme 49. An antibody of any one of Implementation Schemes 1-48, further comprising a TGFβ binding moiety.

[0200] Implementation scheme 50. The antibody of implementation scheme 49, wherein the TGFβ binding portion is linked to the heavy chain variable region.

[0201] Implementation Scheme 51. The antibody of Implementation Scheme 50, wherein the TGFβ binding moiety is linked to the heavy chain variable region via a peptide linker.

[0202] Implementation Scheme 52. An antibody of any one of Implementation Schemes 30-48, further comprising a TGFβ binding moiety linked to the heavy chain constant region.

[0203] Implementation Scheme 53. The antibody of Implementation Scheme 52, wherein the TGFβ binding portion is linked to the C-terminal residue of the heavy chain constant region.

[0204] Implementation Scheme 54. The antibody of Implementation Scheme 52 or 53, wherein the N-terminal residue of the TGFβ binding moiety is linked to the C-terminal residue of the heavy chain constant region via a peptide linker.

[0205] Implementation Scheme 55. The antibody of Implementation Scheme 51 or 54, wherein the peptide linker comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 103-108.

[0206] Implementation Scheme 56. An antibody of any one of Implementation Schemes 49-55, wherein the TGFβ binding portion binds to human TGFβ.

[0207] Implementation Scheme 57. An antibody of any one of Implementation Schemes 49-56, wherein the TGFβ binding portion comprises the extracellular domain of the human TGFβ receptor.

[0208] Implementation scheme 58. The antibody of implementation scheme 57, wherein the human TGFβ receptor is selected from the group consisting of human TGFβR1, TGFβR2 and TGFβR3.

[0209] Implementation scheme 59. The antibody of implementation scheme 57 or 58, wherein the human TGFβ receptor is human TGFβR2.

[0210] Implementation Scheme 60. An antibody of any one of Implementation Schemes 49-59, wherein the TGFβ binding portion comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0211] Implementation Scheme 61. An antibody of any one of Implementation Schemes 49-60, wherein the TGFβ binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0212] Implementation Scheme 62. An antibody of any one of Implementation Schemes 49-61, wherein the TGFβ binding portion comprises an amino acid sequence selected from the group consisting of SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0213] Implementation Scheme 63. An antibody of any one of Implementation Schemes 49-62, wherein the TGFβ binding portion comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0214] Implementation Scheme 64. An antibody of any one of Implementation Schemes 49-63, said antibody comprising a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 113-121, preferably SEQ ID NO: 114 or 115.

[0215] Implementation Scheme 65. An antibody of any one of Implementation Schemes 49-64, wherein the antibody comprises a polypeptide comprising an amino acid sequence selected from SEQ ID NO: 113-121 and a polypeptide comprising an amino acid sequence comprising SEQ ID NO: 92 or 97.

[0216] Implementation Scheme 66. An antibody of any one of Implementation Schemes 49-65, wherein the antibody comprises two polypeptides each comprising an amino acid sequence selected from SEQ ID NO:113-121 and two polypeptides each comprising an amino acid sequence of SEQ ID NO:92 or 97.

[0217] Implementation Scheme 67. An antibody of any one of Implementation Schemes 49-66, wherein the antibody comprises two polypeptides each comprising the amino acid sequence of SEQ ID NO: 115 and two polypeptides each comprising the amino acid sequence of SEQ ID NO: 97.

[0218] Implementation scheme 68. An antibody that binds to human CD73, said antibody comprising a TGFβ binding moiety.

[0219] Implementation Scheme 69. The antibody of Implementation Scheme 68, wherein the antibody is antagonistic to human CD73.

[0220] Implementation Scheme 70. The antibody of Implementation Scheme 69, wherein the antibody deactivates, reduces or inhibits the activity of human CD73.

[0221] Implementation Scheme 71. The antibody of Implementation Scheme 69 or 70, wherein the antibody weakens the ability of human CD73 to convert adenosine monophosphate into adenosine.

[0222] Implementation Scheme 72. An antibody of any one of Implementation Schemes 68-71, wherein the antibody is antagonistic to human CD73 when it binds to TGFβ.

[0223] Implementation Scheme 73. An antibody of any one of Implementation Schemes 68-72, wherein the TGFβ binding portion binds to human TGFβ.

[0224] Implementation Scheme 74. An antibody of any one of Implementation Schemes 68-73, wherein the TGFβ binding portion comprises the extracellular domain of the human TGFβ receptor.

[0225] Implementation Scheme 75. An antibody of any one of Implementation Schemes 68-74, wherein the human TGFβ receptor is selected from the group consisting of human TGFβR1, TGFβR2 and TGFβR3.

[0226] Implementation scheme 76. The antibody of implementation scheme 74 or 75, wherein the human TGFβ receptor is human TGFβR2.

[0227] Implementation Scheme 77. An antibody of any one of Implementation Schemes 68-76, wherein the TGFβ binding portion comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0228] Implementation Scheme 78. An antibody of any one of Implementation Schemes 68-77, wherein the TGFβ binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0229] Implementation Scheme 79. An antibody of any one of Implementation Schemes 68-78, wherein the TGFβ binding portion comprises an amino acid sequence selected from the group consisting of SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0230] Implementation Scheme 80. An antibody of any one of Implementation Schemes 68-79, wherein the TGFβ-binding portion comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0231] Implementation Scheme 81. An antibody of any one of Implementation Schemes 1-48, further comprising a VEGF binding moiety.

[0232] Implementation Scheme 82. The antibody of Implementation Scheme 81, wherein the VEGF binding portion is linked to the heavy chain variable region.

[0233] Implementation Scheme 83. The antibody of Implementation Scheme 82, wherein the VEGF binding portion is linked to the heavy chain variable region via a peptide linker.

[0234] Implementation Scheme 84. An antibody of any one of Implementation Schemes 30-48, further comprising a VEGF binding moiety linked to the heavy chain constant region.

[0235] Implementation Scheme 85. The antibody of Implementation Scheme 84, wherein the VEGF binding portion is linked to the C-terminal residue of the heavy chain constant region.

[0236] Implementation Scheme 86. The antibody of Implementation Scheme 84 or 85, wherein the N-terminal residue of the VEGF binding moiety is linked to the C-terminal residue of the heavy chain constant region via a peptide linker.

[0237] Implementation Scheme 87. The antibody of Implementation Scheme 83 or 86, wherein the peptide linker comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 103-108.

[0238] Implementation scheme 88. An antibody of any one of implementation schemes 81-87, wherein the VEGF binding portion binds to human VEGF.

[0239] Implementation Scheme 89. An antibody of any one of Implementation Schemes 81-88, wherein the VEGF binding portion comprises the extracellular domain of the human VEGF receptor.

[0240] Implementation scheme 90. An antibody of any one of implementation schemes 81-89, wherein the human VEGF receptor is selected from the group consisting of human VEGFR1, VEGFR2 and VEGFR3.

[0241] Implementation scheme 91. The antibody of implementation scheme 89 or 90, wherein the human VEGF receptor is human VEGFR1.

[0242] Implementation Scheme 92. An antibody of any one of Implementation Schemes 81-91, wherein the VEGF binding portion comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the same amino acid sequence as the sequence shown in SEQ ID NO:122.

[0243] Implementation Scheme 93. An antibody of any one of Implementation Schemes 81-92, wherein the VEGF binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO: 122.

[0244] Implementation Scheme 94. An antibody of any one of Implementation Schemes 81-93, wherein the VEGF binding portion comprises the sequence shown in SEQ ID NO: 122.

[0245] Implementation Scheme 95. An antibody of any one of Implementation Schemes 81-94, wherein the VEGF binding portion comprises the sequence shown in SEQ ID NO:122.

[0246] Implementation Scheme 96. An antibody of any one of Implementation Schemes 81-92, said antibody comprising a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NO:123-127.

[0247] Implementation Scheme 97. An antibody of any one of Implementation Schemes 81-96, wherein the antibody comprises a polypeptide comprising an amino acid sequence selected from SEQ ID NO: 123-127 and a polypeptide comprising an amino acid sequence selected from SEQ ID NO: 92, 128 and 97.

[0248] Implementation Scheme 98. An antibody of any one of Implementation Schemes 81-97, wherein the antibody comprises two polypeptides comprising an amino acid sequence selected from SEQ ID NO: 123-127 and two polypeptides comprising an amino acid sequence selected from SEQ ID NO: 92, 128 and 97.

[0249] Implementation scheme 99. An antibody that binds to human CD73, said antibody comprising a VEGF binding moiety.

[0250] Implementation scheme 100. The antibody of implementation scheme 99, wherein the antibody is antagonistic to human CD73.

[0251] Implementation Scheme 101. The antibody of Implementation Scheme 100, wherein the antibody deactivates, reduces or inhibits the activity of human CD73.

[0252] Implementation Scheme 102. The antibody of Implementation Scheme 100, wherein the antibody weakens the ability of human CD73 to convert adenosine monophosphate into adenosine.

[0253] Implementation scheme 103. An antibody of any one of implementation schemes 99-102, wherein the antibody is antagonistic to human CD73 when it binds to VEGF.

[0254] Implementation scheme 104. An antibody of any one of implementation schemes 99-103, wherein the VEGF binding portion binds to human VEGF.

[0255] Implementation scheme 105. An antibody of any one of implementation schemes 99-104, wherein the VEGF binding portion comprises the extracellular domain of the human VEGF receptor.

[0256] Implementation scheme 106. An antibody of any one of implementation schemes 99-105, wherein the human VEGF receptor is selected from the group consisting of human VEGFR1, VEGFR2 and VEGFR3.

[0257] Implementation scheme 107. The antibody of implementation scheme 105 or 106, wherein the human VEGF receptor is human VEGFR1.

[0258] Implementation Scheme 108. An antibody of any one of Implementation Schemes 99-107, wherein the VEGF binding portion comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the same amino acid sequence as the sequence shown in SEQ ID NO:122.

[0259] Implementation Scheme 109. An antibody of any one of Implementation Schemes 99-108, wherein the VEGF binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO:122.

[0260] Implementation Scheme 110. An antibody of any one of Implementation Schemes 99-109, wherein the VEGF binding portion comprises the amino acid sequence shown in SEQ ID NO:122.

[0261] Implementation Scheme 111. An antibody of any one of Implementation Schemes 99-110, wherein the VEGF binding portion consists of the amino acid sequence shown in SEQ ID NO: 122.

[0262] Implementation Scheme 112. The antibody of any of the foregoing implementation schemes further comprises a conjugated cytotoxic agent, cell growth inhibitor, toxin, radionuclide, or detectable marker.

[0263] Implementation Scheme 113. An antibody of any of the foregoing implementation schemes, wherein the antibody binds to an epitope located in a region of human CD73, the amino acid sequence of said region consisting of the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0264] Implementation Scheme 114. An antibody of any of the foregoing implementation schemes, wherein the antibody binds to at least one residue in the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0265] Implementation Scheme 115. An antibody of any of the preceding implementation schemes, wherein the antibody binds to one or more amino acid residues of human CD73 selected from the group consisting of Y158, Y161, P165 or D168 according to the amino acid sequence number of SEQ ID NO:99.

[0266] Implementation Scheme 116. An antibody of any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 133 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0267] Implementation Scheme 117. An antibody according to any of the foregoing embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:134 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0268] Implementation Scheme 118. An antibody of any of the preceding implementation schemes, wherein the antibody binds to amino acid residue Y158 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0269] Implementation Scheme 119. An antibody according to any of the foregoing implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:59 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0270] Implementation Scheme 120. An antibody of any of the preceding implementation schemes, wherein the antibody binds to amino acid residue Y161 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0271] Implementation Scheme 121. The antibody of any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 60 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0272] Implementation Scheme 122. The antibody of any of the preceding implementation schemes, wherein the antibody binds to amino acid residue P165 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0273] Implementation Scheme 123. The antibody of any of the foregoing implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:139 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0274] Implementation Scheme 124. An antibody of any of the preceding implementation schemes, wherein the antibody binds to amino acid residue D168 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0275] Implementation Scheme 125. An antibody according to any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 140 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0276] Implementation Scheme 126. The antibody of any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:135 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0277] Implementation Scheme 127. The antibody of any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 136 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0278] Implementation Scheme 128. An antibody according to any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 137 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0279] Implementation Scheme 129. An antibody according to any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 138 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0280] Implementation Scheme 130. An antibody that binds to human CD73, wherein the antibody binds to an epitope located in a region of human CD73, the amino acid sequence of said region consisting of the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0281] Implementation Scheme 131. An antibody that binds to human CD73, wherein the antibody binds to at least one residue in the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0282] Implementation Scheme 132. An antibody that binds to human CD73, wherein the antibody binds to one or more amino acid residues of human CD73 selected from the group consisting of Y158, Y161, P165 or D168 according to the amino acid sequence number of SEQ ID NO:99.

[0283] Implementation Scheme 133. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 133 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0284] Implementation Scheme 134. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 134 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0285] Implementation scheme 135. An antibody that binds to human CD73, wherein the antibody binds to amino acid residue Y158 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0286] Implementation Scheme 136. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:59 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0287] Implementation scheme 137. An antibody that binds to human CD73, wherein the antibody binds to amino acid residue Y161 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0288] Implementation Scheme 138. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 60 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0289] Implementation scheme 139. An antibody that binds to human CD73, wherein the antibody binds to amino acid residue P165 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0290] Implementation Scheme 140. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:139 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO:141.

[0291] Implementation Scheme 141. An antibody that binds to human CD73, wherein the antibody binds to amino acid residue D168 of human CD73 according to the amino acid sequence number of SEQ ID NO:99.

[0292] Implementation Scheme 142. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 140 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0293] Implementation Scheme 143. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 135 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0294] Implementation Scheme 144. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 136 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0295] Implementation Scheme 145. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 137 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0296] Implementation Scheme 146. An antibody that binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 138 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0297] Implementation scheme 147. A compound or molecule comprising a CD73 binding moiety and a TGFβ binding moiety.

[0298] Implementation scheme 148. A polynucleotide encoding a polypeptide of an antibody of any one of implementation schemes 1-146.

[0299] Implementation Scheme 149. The polynucleotide of Implementation Scheme 148, wherein the polynucleotide comprises cDNA or mRNA.

[0300] Implementation scheme 150. A vector comprising the polynucleotides of implementation scheme 148 or 149.

[0301] Implementation scheme 151. The vector of implementation scheme 150, wherein the vector is a plasmid vector or a viral vector.

[0302] Implementation scheme 152. A recombinant host cell containing the polynucleotide of implementation scheme 148 or 149 or the vector of implementation scheme 150 or 151.

[0303] Implementation Scheme 153. Lipid nanoparticles comprising the polynucleotide of Implementation Scheme 148 or 149 or the carrier of Implementation Scheme 150 or 151.

[0304] Implementation Scheme 154. A pharmaceutical composition comprising an antibody, polynucleotide, carrier or lipid nanoparticle, and a pharmaceutically acceptable carrier or excipient of any one of Implementation Schemes 1-146 or 148-153.

[0305] Implementation Scheme 155. A method for generating antibodies, the method comprising culturing a host cell of Implementation Scheme 152 under suitable conditions to express the polynucleotide and generate the antibody.

[0306] Implementation Scheme 156. A method for treating cancer in a subject, the method comprising administering to the subject an effective amount of an antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153, or 154.

[0307] Implementation Scheme 157. A method for inhibiting cancer metastasis in a subject, the method comprising administering to the subject an effective amount of an antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153, or 154.

[0308] Implementation scheme 158. The method of implementation scheme 156 or 157, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered intravenously, intratumorally, subcutaneously, intradermally, intramuscularly, intrabladderally, intracranially, intracavitarily or intraventricularly.

[0309] Implementation scheme 159. The method of implementation scheme 156 or 157, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered intratumorally.

[0310] Implementation scheme 160. The method of any one of implementation schemes 156-159, further comprising administering one or more additional therapeutic agents to the subject.

[0311] Implementation Scheme 161. The method of any one of Implementation Schemes 156-160, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition and the one or more additional therapeutic agents are co-administered according to the same schedule (e.g., co-administered at the same time intervals).

[0312] Implementation Scheme 162. The method of any one of Implementation Schemes 156-160, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition and one or more additional therapeutic agents are co-administered according to different schedules (e.g., co-administered at different time intervals).

[0313] Implementation Scheme 163. The method of any one of Implementation Schemes 156-162, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered in combination with one or more additional therapeutic agents, said one or more additional therapeutic agents comprising an activator or agonist of an FMS-associated tyrosine kinase 3 (FLT3; CD135) receptor, a Toll-like receptor (TLR) or an interferon gene stimulator (STING) receptor.

[0314] Implementation Scheme 164. The method of Implementation Scheme 163, wherein the TLR agonist or activator is selected from the group consisting of: TLR2 agonists, TLR3 agonists, TLR7 agonists, TLR8 agonists and TLR9 agonists.

[0315] Implementation Scheme 165. The method of Implementation Scheme 163, wherein the STING receptor agonist or activator is selected from the group consisting of: ADU-S100 (MIW-815), SB-11285, MK-1454, SR-8291, AdVCA0848, GSK-532, SYN-STING, MSA-1, SR-8291, 5,6-dimethylxanthenone-4-acetic acid (DMXAA), cyclic GAMP (cGAMP), and cyclic diAMP.

[0316] Implementation Scheme 166. The method of any one of Implementation Schemes 156-165, wherein the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is co-administered with one or more additional therapeutic agents, said one or more additional therapeutic agents comprising inhibitors or antagonists of: non-receptor protein tyrosine phosphatase 11 (PTPN11 or SHP2), myeloid leukemia sequence 1 (MCL1) apoptosis regulator; mitogen-activated protein kinase kinase 1 (MAP4K1) (also known as hematopoietic progenitor cell kinase 1 (HPK1)); diacylglycerol kinase alpha (DGKA, DAGK, DAGK1, or DGK-alpha); extracellular 5'-nucleotidase (NT5E or CD73); transforming growth factor beta 1 (TGFB1 or TGFβ); heme oxygenase 1 (HMOX1, HO-1, or HO1); vascular endothelial growth factor A (VEGFA or VEGF); erb-b2 receptor tyrosine kinase 2 (ERBB2). HER2, HER2 / neu, or CD340; epidermal growth factor receptor (EGFR, ERBB, ERBB1, or HER1); ALK receptor tyrosine kinase (ALK, CD246); poly(ADP-ribose) polymerase 1 (PARP1 or PARP); cyclin-dependent kinase 4 (CDK4); cyclin-dependent kinase 6 (CDK6); CC motif chemokine receptor 8 (CCR8, CDw198); CD274 molecule (CD274, PDL1, or PD-L1); programmed cell death 1 (PDCD1, PD1, or PD-1); and / or cytotoxic T lymphocyte-associated protein 4 (CTLA4, CTLA-4, CD152).

[0317] Implementation Scheme 167. The method of Implementation Scheme 166, wherein the inhibitor comprises an antigen-binding molecule, an antibody, or an antigen-binding fragment thereof.

[0318] Implementation scheme 168. The method of implementation scheme 166, wherein the inhibitor comprises small organic molecules.

[0319] Implementation scheme 169. The method of implementation scheme 166, wherein the MCL1 inhibitor is selected from the following group: AMG-176, AMG-397, S-64315, AZD-5991, 483-LM, A1210477, UMI-77 and JKY-5-037.

[0320] Implementation scheme 170. The method of implementation scheme 166, wherein the inhibitor of PTPN11 or SHP2 is selected from the group consisting of: TNO155 (SHP-099), RMC-4550, JAB-3068 and RMC-4630.

[0321] Implementation Scheme 171. The method of Implementation Scheme 160, wherein the additional therapeutic agent is a chemotherapeutic agent, an anti-biological agent, a radiotherapy agent, or a checkpoint targeting agent.

[0322] Implementation Scheme 172. The method of Implementation Scheme 160, wherein the one or more anti-biological agents or chemotherapeutic agents are selected from the group consisting of: nucleoside analogs (e.g., 5-fluorouracil, gemcitabine, cytarabine), taxanes (e.g., paclitaxel, nab-paclitaxel, docetaxel, cabazitaxel), platinum complexes (cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin).tetranitrate, phenanthriplatin, picoplatin, satraplatin, dicycloplatin, eptaplatin, lobaplatin, miriplatin, dihydrofolate reductase (DHFR) inhibitors (e.g., methotrexate, trimetrexate, pemetrexed), topoisomerase inhibitors (e.g., doxorubicin, daunorubicin, dactinomycin, eniposide, epirubicin, etoposide, idarubicin, irinotecan, mitoxantrone, pixantrone, sobuzoxane, topotecan otecan), irinotecan, MM-398 (irinotecan liposome), vosaroxin and GPX-150, aldoxorubicin, AR-67, growlertinib, AST-2818, avitinib (ACEA-0010), irofulven (MGI-114), alkylating agents (e.g., nitrogen mustard, cyclophosphamide, chlormethine, urethane) Uramustine or uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, temozolomide, carmustine, nitrosoureas (e.g., carmustine, lomustine, streptozocin), alkyl sulfonates (e.g., busulfan), and mixtures thereof.

[0323] Implementation Scheme 173. The method of Implementation Scheme 160, wherein the checkpoint targeting agent is selected from the group consisting of: antagonist anti-PD-1 antibody, antagonist anti-PD-L1 antibody, antagonist anti-PD-L2 antibody, antagonist anti-CTLA-4 antibody, antagonist anti-TIM-3 antibody, antagonist anti-LAG-3 antibody, antagonist anti-CEACAM1 antibody, agonist anti-GITR antibody, antagonist anti-TIGIT antibody, antagonist anti-VISTA antibody, agonist anti-CD137 antibody, and agonist anti-OX40 antibody.

[0324] Implementation Scheme 174. The method of Implementation Scheme 173, wherein the additional therapeutic agent is an anti-PD-1 antibody, optionally wherein the anti-PD-1 antibody is pembrolizumab or nivolumab.

[0325] Implementation Scheme 175. The method of Implementation Scheme 160, wherein the additional therapeutic agent is an inhibitor of indoleamine-2,3-dioxygenase (IDO).

[0326] Implementation Scheme 176. The method of Implementation Scheme 175, wherein the inhibitor is selected from the group consisting of: epacadostat, F001287, indoximod and NLG919.

[0327] Implementation scheme 177. The method of implementation scheme 176, wherein the inhibitor is edodestat.

[0328] Implementation scheme 178. The method of implementation scheme 160, wherein the additional therapeutic agent is a vaccine.

[0329] Implementation Scheme 179. The method of Implementation Scheme 178, wherein the vaccine comprises a heat shock peptide complex (HSPPC), the HSPPC comprising a heat shock protein complexed with an antigenic peptide.

[0330] Implementation scheme 180. The method of implementation scheme 179, wherein the heat shock protein is hsc70 and is complexed with a tumor-associated antigenic peptide.

[0331] Implementation Scheme 181. The method of Implementation Scheme 179, wherein the heat shock protein is gp96 and is complexed with a tumor-associated antigenic peptide, wherein the HSPPC is derived from a tumor obtained from the subject.

[0332] Implementation Scheme 182. The method of Implementation Scheme 160, wherein the additional therapeutic agent is etoposide or doxorubicin.

[0333] Implementation Scheme 183. The method of any one of Implementation Schemes 156-182, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered in combination with the FOLFOX regimen, the FOLFOXIRI regimen or the FOLFIRINOX regimen.

[0334] Implementation scheme 184. The method of any one of implementation schemes 156-183, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered in combination with immunotherapy, immunostimulation therapy, cell therapy or gene therapy.

[0335] Implementation scheme 185. The method of any one of implementation schemes 156-184, wherein the cancer is selected from the group consisting of solid tumors, hematologic cancers and metastatic lesions.

[0336] Implementation scheme 186. The method of implementation scheme 185, wherein the solid tumor is selected from the group consisting of sarcoma, fibroblastic sarcoma, carcinoma and adenocarcinoma.

[0337] Implementation scheme 187. The method of implementation scheme 185, wherein the hematologic cancer is selected from the group consisting of leukemia, lymphoma and myeloma.

[0338] Implementation Scheme 188. The method of any one of Implementation Schemes 156-184, wherein the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloid monocytic leukemia, etc. Nucleated cell leukemia (CMML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma.

[0339] Implementation Scheme 189. The method of any one of Implementation Schemes 156-184, wherein the cancer is selected from the group consisting of: epithelial tumors (e.g., carcinoma, squamous cell carcinoma, basal cell carcinoma, squamous intraepithelial neoplasm), adenomatous tumors (e.g., adenocarcinoma, adenoma, adenomyoma), stromal or soft tissue tumors (e.g., sarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, fibrosarcoma, dermatofibrosarcoma, neurofibrosarcoma, fibrous histiocytoma, angiosarcoma, angiomyosarcoma, leiomyosarcoma, chondroma, chondrosarcoma, alveolar soft sarcoma, epithelioid angioendothelioma, Spitz tumor, synovial sarcoma), and lymphoma.

[0340] Implementation Scheme 190. The method of any one of Implementation Schemes 156-184, wherein the cancer is a solid tumor in or originating from a tissue or organ selected from the group consisting of: bone (e.g., ameloblastoma, aneurysmal bone cyst, angiosarcoma, chondroblastoma, chondroma, chondromycinoma, chondrosarcoma, chordoma, dedifferentiated chondrosarcoma, enchondroma, epithelioid hemangioendothelioma, fibrous dysplasia of bone, giant cell tumor of bone, hemangioma and related lesions, osteoblastoma, osteochondroma, osteosarcoma, osteoid osteoma, osteoma, periosteal chondroma, desmoidoma, Ewing sarcoma); lips and oral cavity (e.g., odontogenic ameloblastoma, oral leukoplakia, oral squamous cell carcinoma, primary ameloblastoma, etc.). Primary oral mucosal melanoma; salivary glands (e.g., pleomorphic salivary gland adenoma, salivary gland adenoid cystic carcinoma, salivary gland mucoepidermoid carcinoma, salivary gland Warthin tumor); esophagus (e.g., Barrett's esophagus, dysplasia, and adenocarcinoma); gastrointestinal tract, including the stomach (e.g., gastric adenocarcinoma, primary gastric lymphoma, gastrointestinal stromal tumor (GIST), metastatic deposits, gastric carcinoid, gastric sarcoma, neuroendocrine carcinoma, primary squamous cell carcinoma of the stomach, gastric adenoacanthoma), intestine and smooth muscle (e.g., intravenous leiomyomatosis), colon (e.g., colorectal adenocarcinoma), rectum, anus; pancreas (e.g., serous neoplasms, including microcystic or macrocystic serous cystadenomas, solid serous cystadenomas, Von Hippel-Landau (VHL) related serous cystic neoplasms, serous cystadenocarcinoma, mucinous cystic neoplasms (MCN), intraductal papillary mucinous neoplasms (IPMN), intraductal eosinophilic papillary neoplasms (IOPN), intraductal tubular neoplasms, cystic acinar neoplasms including acinar cell cystadenoma, acinar cell cystadenocarcinoma, pancreatic adenocarcinoma, invasive pancreatic ductal adenocarcinoma including tubular adenocarcinoma, adenosquamous carcinoma, colloid carcinoma, medullary carcinoma, hepatoid carcinoma, signet ring cell carcinoma, undifferentiated carcinoma, undifferentiated carcinoma with osteoclast-like giant cells, acinar cell carcinoma, neuroendocrine neoplasms Biological organisms, including neuroendocrine microadenomas, neuroendocrine tumors (NETs), neuroendocrine carcinomas (NECs), including small cell or large cell NECs, insulinomas, gastrinomas, glucagonomas, serotonin-producing NETs, ​​somatostatinomas, angiotensinomas, solid pseudopapillary neoplasms (SPNs), pancreatoblastomas; gallbladder (e.g., gallbladder cancer and extrahepatic bile duct cancer, intrahepatic bile duct cancer); neuroendocrine (e.g., adrenocortical carcinoma, carcinoid tumors, pheochromocytomas, pituitary adenomas); thyroid (e.g., anaplastic (undifferentiated) carcinoma, medullary carcinoma, eosinophilic tumors, papillary carcinoma, adenocarcinoma);Liver (e.g., adenoma, combined hepatocellular and cholangiocarcinoma, fibrolamellar carcinoma, hepatoblastoma, hepatocellular carcinoma, mesenchymal, nested stromal epithelial tumors, undifferentiated carcinoma, hepatocellular carcinoma, intrahepatic cholangiocarcinoma, bile duct cystadenocarcinoma, epithelioid hemangioendothelioma, angiosarcoma, embryonal sarcoma, rhabdomyosarcoma, solitary fibroma, teratoma, yolk sac tumor, carcinosarcoma, rhabdomyosarcoma); kidney (e.g., ALK rearranged renal cell carcinoma, chromophobe renal cell carcinoma, clear cell renal cell carcinoma, clear cell sarcoma, metanephrotic adenoma, metanephrotic fibroma, mucinous tubular and spindle cell carcinoma, nephrotic tumor, nephroblastoma). Wilms' tumor, papillary adenoma, papillary renal cell carcinoma, renal eosinophilic cell carcinoma, renal cell carcinoma, succinate dehydrogenase-deficient renal cell carcinoma, collecting duct carcinoma); breast (e.g., invasive ductal carcinoma, including but not limited to acinic cell carcinoma, adenoid cystic carcinoma, apocrine gland carcinoma, cribriform carcinoma, glycogen-rich / clear cell carcinoma, inflammatory carcinoma, lipid-rich carcinoma, medullary carcinoma, metaplastic carcinoma, micropapillary carcinoma, mucinous carcinoma, neuroendocrine carcinoma, eosinophilic carcinoma, papillary carcinoma, sebaceous gland carcinoma, secretory breast cancer, tubular carcinoma, lobular carcinoma, including but not limited to pleomorphic carcinoma, immature carcinoma, etc.). Cellular carcinomas, peritoneum (e.g., mesothelioma, primary peritoneal carcinoma); female sex organ tissues, including ovaries (e.g., choriocarcinoma, epithelial tumors, germ cell tumors, sex cord-stromal tumors), fallopian tubes (e.g., serous adenocarcinoma, mucinous adenocarcinoma, endometrioid adenocarcinoma, clear cell adenocarcinoma, transitional cell carcinoma, squamous cell carcinoma, undifferentiated carcinoma, Müllerian tumor, adenosarcoma, leiomyosarcoma, teratoma, germ cell tumors, choriocarcinoma, trophoblastic tumors), uterus (e.g., cervical cancer, endometrial polyps, endometrial hyperplasia, epithelial intraepithelial carcinoma (EIC), endometrial cancer). (e.g., endometrioid carcinoma, serous carcinoma, clear cell carcinoma, mucinous carcinoma, squamous cell carcinoma, transitional carcinoma, small cell carcinoma, undifferentiated carcinoma, mesenchymal neoplasms), leiomyomas (e.g., endometrial stromal nodules, leiomyosarcoma, endometrial stromal sarcoma (ESS), mesenchymal tumors), mixed epithelial and mesenchymal tumors (e.g., adenofibroma, carcinoma fibroma, adenosarcoma, carcinoma sarcoma (malignant mixed mesodermal sarcoma - MMMT), endometrial stromal tumors, malignant mixed Müllerian tumors of the endometrium, gestational trophoblastic tumors (partial hydatiform mole) Mole, complete hydatidiform mole, invasive hydatidiform mole, placental site tumors); vulva, vagina; male sex organs, including prostate, testes (e.g., germ cell tumors, seminoma), penis; bladder (e.g., squamous cell carcinoma, urothelial carcinoma, bladder urothelial carcinoma); brain (e.g., gliomas (e.g., astrocytomas, including non-invasive, low-grade, anaplastic, glioblastomas; oligodendrogliomas, ependymomas), meningiomas, gangliogliomas, Schwann cell tumors (schwannomas), craniopharyngiomas, chordomas, non-Hodgkin lymphomas, pituitary tumors);Eye (e.g., retinoma, retinoblastoma, ocular melanoma, retrouvian melanoma, iris hamartoma); head and neck (e.g., nasopharyngeal carcinoma, endolymphatic cystic tumor (ELST), epidermoid carcinoma, laryngeal carcinoma including squamous cell carcinoma (SCC) (e.g., glottic carcinoma, supraglottic carcinoma, subglottic carcinoma, transglottic carcinoma), carcinoma in situ, verrucous, spindle cell and basaloid SCC, undifferentiated carcinoma, laryngeal adenocarcinoma, adenoid cystic carcinoma, neuroendocrine carcinoma, laryngeal sarcoma), head and neck paraganglioma (e.g., carotid body, tympanic cavity, vagus nerve); thymus (e.g., thymoma); heart (e.g., cardiac myxoma); lung (e.g., small cell carcinoma (SCLC), non-small cell lung cancer (NSCLC), including squamous cell carcinoma (SCC), adenocarcinoma and large cell carcinoma, carcinoid (typical or atypical), carcinosarcoma, pulmonary blastoma, giant cell carcinoma). Spindle cell carcinoma, pleural pulmonary blastoma; lymphoma (e.g., lymphoma, including Hodgkin's lymphoma, non-Hodgkin's lymphoma, EBV-associated lymphoproliferative disorders, including B-cell lymphoma and T-cell lymphoma (e.g., Burkitt lymphoma, large B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, indolent B-cell lymphoma, low-grade B-cell lymphoma, fibrin-associated diffuse large cell lymphoma; primary exudative lymphoma; plasmablastic lymphoma; extranodal NK / T-cell lymphoma (nasal type); peripheral T-cell lymphoma, cutaneous T-cell lymphoma, angioimmunoblastic T-cell lymphoma; follicular T-cell lymphoma; systemic T-cell lymphoma, lymphangioleiomyomatosis);Central nervous system (CNS) tumors (e.g., gliomas, including astrocytic tumors such as pilocytic astrocytoma, pilocyxoid astrocytoma, subependymal giant cell astrocytoma, pleomorphic xanthoastrocytoma, diffuse astrocytoma, fibrous astrocytoma, and gemistocytic astrocytoma). astrocytoma, protoplasmic astrocytoma, anaplastic astrocytoma, glioblastoma (e.g., giant cell glioblastoma, glioma, glioblastoma multiforme, and gliomatosis), oligodendroglioma (e.g., oligodendroglioma, anaplastic oligodendroglioma), oligodendroastrocytic tumor (e.g., oligodendroglioma, anaplastic oligodendroglioma), ependymal tumor (e.g., subependymoma, myxopapillary ependymoma, ependymoma (e.g., cellular, papillary, clear cell, elongated cell), anaplastic ependymoma), optic glioma and non-glial tumor (e.g., choroid plexus tumors, neuronal and mixed neuronal glial tumors, pineal region tumors, embryonal tumors). Tumors, including medulloblastomas, meningeal tumors, primary CNS lymphomas, germ cell tumors, pituitary adenomas, cranial nerve and paravertebral nerve tumors, stellate region tumors, neurofibromas, meningiomas, peripheral nerve sheath tumors, peripheral neuroblastomas (including but not limited to neuroblastomas, ganglioneuromas, ganglioneuromas), trisomy 19 ependymomas; neuroendocrine tissues (e.g., paraganglionic systems, including adrenal medulla (pheochromocytoma) and extra-adrenal paragangliomas (extra-adrenal paragangliomas); skin (e.g., clear cell hidradenoma, benign fibrous histiocytoma of the skin, cylindrica, hidradenoma, melanoma (including cutaneous melanoma, mucosal melanoma), pilomatoma, Spitz tumors);and soft tissue (e.g., aggressive angiomyxoma, alveolar rhabdomyosarcoma, alveolar soft sarcoma, angiofibroma, hemangioma-like fibrous histiocytoma, synovial sarcoma, biphasic synovial sarcoma, clear cell sarcoma, dermatofibrosarcoma protuberans, desmoid fibromatosis, small round cell tumor, desmoplastic small round cell tumor, elastic fibroma, embryonal rhabdomyosarcoma, Ewing's tumor / primary neuroectodermal tumor (PNET), bone) Extrailiac myxoid chondrosarcoma, extraskeletal osteosarcoma, paravertebral sarcoma, inflammatory myofibroblastic tumor, lipoblastoma, lipoma, chondroid lipoma, liposarcoma / malignant lipomatous tumor, liposarcoma, myxoid liposarcoma, fibromyxoid sarcoma, lymphangioleiomyosarcoma, malignant myoepithelioma, soft tissue malignant melanoma, myoepithelial carcinoma, myoepithelioma, myxoid inflammatory fibroblastic sarcoma, undifferentiated sarcoma, peridermoid cell tumor, rhabdomyosarcoma, non-rhabdomyosarcoma soft tissue sarcoma, soft tissue leiomyosarcoma, undifferentiated sarcoma, well-differentiated liposarcoma.

[0341] Implementation Scheme 191. The method of any one of Implementation Schemes 156-184, wherein the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal cancer, and endometrial cancer.

[0342] Implementation scheme 192. An antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any one of implementation schemes 1-146, 148-151, 153 or 154, for the treatment of cancer.

[0343] Implementation scheme 193. The antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition used in implementation scheme 192, wherein the cancer is selected from the group consisting of solid tumors, hematologic cancers and metastatic lesions.

[0344] Implementation scheme 194. The antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition used in implementation scheme 192, wherein the solid tumor is selected from the group consisting of sarcoma, fibroblastic sarcoma, carcinoma and adenocarcinoma.

[0345] Implementation scheme 195. The antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition used in implementation scheme 192, wherein the hematologic cancer is selected from the group consisting of leukemia, lymphoma and myeloma.

[0346] Implementation Scheme 196. The antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition used in any of Implementation Schemes 192-195, wherein the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and chronic myeloid leukemia. Chronic myeloid monocytic leukemia (CML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma.

[0347] Implementation Scheme 197. An antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition of any one of Implementation Schemes 192-195, wherein the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal cancer, and endometrial cancer.

[0348] Implementation Scheme 198. Use of any antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any of Implementation Schemes 1-146, 148-151, 153 or 154 in the preparation of a medicament for the treatment of cancer.

[0349] Implementation scheme 199. Use of implementation scheme 198, wherein the cancer is selected from the group consisting of solid tumors, hematologic cancers and metastatic lesions.

[0350] Implementation scheme 200. Use of implementation scheme 199, wherein the solid tumor is selected from the group consisting of sarcoma, fibroblastic sarcoma, carcinoma and adenocarcinoma.

[0351] Implementation scheme 201. Use of implementation scheme 199, wherein the hematologic cancer is selected from the group consisting of leukemia, lymphoma and myeloma.

[0352] Implementation Scheme 202. Use of any one of Implementation Schemes 198-201, wherein the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloid monocytic leukemia, etc. Nucleated cell leukemia (CMML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma.

[0353] Implementation Scheme 203. Use of any one of Implementation Schemes 198-201, wherein the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal cancer, and endometrial cancer.

[0354] Implementation Scheme 204. An antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153 or 154, used in the method of any one of Implementation Schemes 156-191.

[0355] Implementation Scheme 205. Use of any antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any of Implementation Schemes 1-146, 148-151, 153 or 154 in the method of any of Implementation Schemes 156-191.

[0356] This application relates to the following implementation schemes:

[0357] Implementation Scheme 1. An isolated antibody that specifically binds to human CD73, said antibody comprising: a heavy chain variable region containing complementarity-determining regions (CDRs) CDRH1, CDRH2, and CDRH3, and a light chain variable region containing complementarity-determining regions CDRL1, CDRL2, and CDRL3, wherein:

[0358] (a) CDRH1 contains the amino acid sequence X1X2WX3X4 (SEQ ID NO:13), in which

[0359] X1 is S or N;

[0360] X2 is either S or Y;

[0361] X3 is either I or M; and

[0362] X4 is N or H;

[0363] (b) CDRH2 contains the amino acid sequence X1IYPRX2X3DTNYX4X5KFKX6 (SEQ ID NO:14), in which

[0364] X1 is either R or T;

[0365] X2 is N, A, or S;

[0366] X3 is either G or S;

[0367] X4 is N, A, or S;

[0368] X5 is either G or Q; and

[0369] X6 is either D or G;

[0370] (c) CDRH3 contains the amino acid sequence of LLDYSMDY (SEQ ID NO:7);

[0371] (d) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0372] X1 is N or I; and

[0373] X2 is either Y or S;

[0374] (e) CDRL2 contains the amino acid sequence YTSRLHS (SEQ ID NO: 10); and / or

[0375] (f) CDRL3 contains the amino acid sequence QQGNTLPXT (SEQ ID NO:17), wherein:

[0376] X is either L or W.

[0377] Implementation Scheme 2. The isolated antibody of Implementation Scheme 1, wherein CDRH1 contains an amino acid sequence selected from the group consisting of SEQ ID NO: 1 and 2.

[0378] Implementation Scheme 3. The isolated antibody of Implementation Scheme 1 or 2, wherein CDRH2 contains an amino acid sequence selected from the group consisting of: SEQ ID NO: 3-6.

[0379] Implementation Scheme 4. The isolated antibody of any of the foregoing implementation schemes, wherein CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 8 and 9.

[0380] Implementation Scheme 5. The isolated antibody of any of the preceding implementation schemes, wherein CDRL3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 11 and 12.

[0381] Implementation Scheme 6. The isolated antibody from any of the aforementioned implementation schemes, wherein:

[0382] (a) CDRH1 contains the amino acid sequence of SSWIN (SEQ ID NO:2);

[0383] (b) CDRH2 contains the amino acid sequence RIYPRX1GDTNYX2GKFKD (SEQ ID NO:15), in which

[0384] X1 is N, A, or S; and

[0385] X2 is N, A, or S;

[0386] (c) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0387] X1 is N or I; and

[0388] X2 is Y or S; and / or

[0389] (d)CDRL3 contains the amino acid sequence QQGNTLPLT (SEQ ID NO:12).

[0390] Implementation Scheme 7. The isolated antibody of Implementation Scheme 6, wherein CDRH2 contains an amino acid sequence selected from the group consisting of: SEQ ID NO:4-6.

[0391] Implementation scheme 8. The isolated antibody of implementation scheme 6 or 7, wherein CDRL1 contains an amino acid sequence selected from the group consisting of SEQ ID NO: 8 and 9.

[0392] Implementation Scheme 9. An isolated antibody from any of the preceding implementation schemes, wherein CDRH1, CDRH2 and CDRH3 respectively contain the amino acid sequences of CDRH1, CDRH2 and CDRH3 shown in SEQ ID NO: 2, 4 and 7; 2, 5 and 7; 1, 3 and 7; or 2, 6 and 7.

[0393] Implementation Scheme 10. An isolated antibody from any of the preceding implementation schemes, wherein CDRL1, CDRL2 and CDRL3 respectively comprise the amino acid sequences of CDRL1, CDRL2 and CDRL3 shown in SEQ ID NO: 8, 10 and 12; 9, 10 and 12; or 8, 10 and 11.

[0394] Implementation Scheme 11. The isolated antibody of any of the preceding implementation schemes, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 respectively comprise SEQ ID NO: 2, 4, 7, 8, 10 and 12; 2, 5, 7, 8, 10 and 12; 1, 3, 7, 8, 10 and 11; 2, 4, 7, 8, 10 and 11; 2, 4, 7, 9, 10 and 12; 2, 6, 7, 8, 10 and 12; 2, 5, 7, 9, 10 and 12; or the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 shown in 2, 6, 7, 9, 10 and 12.

[0395] Implementation Scheme 12. The isolated antibodies of Implementation Scheme 11, wherein CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 respectively contain SEQ ID NO: 2, 4, 7, 8, 10 and 12; or the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2 and CDRL3 shown in 2, 5, 7, 8, 10 and 12.

[0396] Implementation Scheme 13. An isolated antibody that specifically binds to human CD73, said antibody comprising a heavy chain variable region containing CDR CDRH1, CDRH2 and CDRH3, and a light chain variable region containing complementarity-determining regions CDRL1, CDRL2 and CDRL3, wherein CDRH3 contains the amino acid sequence LLDYSMDY (SEQ ID NO:7); and / or CDRL3 contains the amino acid sequence QQGNTLPLT (SEQ ID NO:12) or QQGNTLPWT (SEQ ID NO:11).

[0397] Implementation Scheme 14. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31.

[0398] Implementation Scheme 15. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 19-21 and 23-30.

[0399] Implementation Scheme 16. The isolated antibody of Implementation Scheme 15, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 19-21 and 23-30.

[0400] Implementation Scheme 17. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of: SEQ ID NO: 23-30.

[0401] Implementation Scheme 18. The isolated antibody of Implementation Scheme 17, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:23-30.

[0402] Implementation Scheme 19. The isolated antibody of Implementation Scheme 18, wherein the heavy chain variable region comprises the amino acid sequence of SEQ ID NO: 24 or 27.

[0403] Implementation Scheme 20. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain variable region comprising an amino acid sequence derived from the human lineage sequence shown in SEQ ID NO:22.

[0404] Implementation Scheme 21. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a light chain variable region comprising the amino acid sequence of SEQ ID NO: 41 or 42.

[0405] Implementation Scheme 22. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a light chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 33-35 and 37-40.

[0406] Implementation Scheme 23. The isolated antibody of Implementation Scheme 22, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 33-35 and 37-40.

[0407] Implementation Scheme 24. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a light chain variable region comprising an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to an amino acid sequence selected from the group consisting of: SEQ ID NO:37-40.

[0408] Implementation Scheme 25. The isolated antibody of Implementation Scheme 24, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:37-40.

[0409] Implementation Scheme 26. The isolated antibody of Implementation Scheme 25, wherein the light chain variable region comprises the amino acid sequence of SEQ ID NO:37.

[0410] Implementation Scheme 27. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a light chain variable region comprising an amino acid sequence derived from the human lineage sequence shown in SEQ ID NO:36.

[0411] Implementation Scheme 28. An isolated antibody from any of the preceding implementation schemes, wherein the heavy chain variable region and the light chain variable region comprise, respectively, the amino acid sequences shown in SEQ ID NO: 24 and 37; 27 and 37; 19 and 33; 20 and 34; 21 and 35; 23 and 37; 23 and 38; 24 and 38; 25 and 37; 26 and 37; 28 and 37; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 29 and 37; 30 and 37; 23 and 39; or 23 and 40.

[0412] Implementation Scheme 29. The isolated antibody of Implementation Scheme 28, wherein the heavy chain variable region and the light chain variable region comprise the amino acid sequences shown in SEQ ID NO: 24 and 37, respectively; or 27 and 37.

[0413] Implementation Scheme 30. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain constant region selected from the group consisting of human IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2.

[0414] Implementation Scheme 31. The isolated antibody of Implementation Scheme 30, wherein the heavy chain constant region is IgG1.

[0415] Implementation Scheme 32. The isolated antibody of Implementation Scheme 31, wherein the amino acid sequence of IgG1 contains the N297A mutation numbered according to the EU numbering system.

[0416] Implementation Scheme 33. The isolated antibody of Implementation Scheme 32, wherein the heavy chain constant region comprises the amino acid sequence of SEQ ID NO: 50, 49, 46, 45, 55 or 56.

[0417] Implementation Scheme 34. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain comprising an amino acid sequence selected from the group consisting of: SEQ ID NO:63-88.

[0418] Implementation Scheme 35. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a light chain constant region selected from the group consisting of human Igκ and Igλ.

[0419] Implementation scheme 36. The isolated antibody of implementation scheme 35, wherein the light chain constant region is Igκ.

[0420] Implementation Scheme 37. The isolated antibody of Implementation Scheme 36, wherein the light chain constant region comprises the amino acid sequence of SEQ ID NO: 93 or 89.

[0421] Implementation Scheme 38. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a light chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 92 and 94-98.

[0422] Implementation Scheme 39. An isolated antibody from any of the preceding implementation schemes, wherein the antibody comprises a heavy chain and a light chain, the heavy chain and the light chain each comprising an amino acid sequence shown in any row of Table 4, preferably selected from the group consisting of: SEQ ID NO: 79 and 97; 85 and 97; 77 and 97; 80 and 97; 86 and 97; 64 and 92; 70 and 96; or 78 and 97.

[0423] Implementation Scheme 40. An antibody isolated from any one of Implementation Schemes 15-39, wherein X is glutamine in any one of SEQ ID NO: 19, 20 and 65-68.

[0424] Implementation Scheme 41. An antibody isolated from any one of Implementation Schemes 15-39, wherein X is glutamic acid in any one of SEQ ID NO: 21, 23-30, 63, 64, 69-88.

[0425] Implementation Scheme 42. An antibody isolated from any one of Implementation Schemes 15-39, wherein X is pyroglutamic acid in any one of SEQ ID NO: 19-21, 23-30 and 63-88.

[0426] Implementation Scheme 43. An isolated antibody that binds to the same human CD73 epitope as the antibody in any of the preceding implementation schemes.

[0427] Implementation Scheme 44. The isolated antibody of any of the foregoing implementation schemes, wherein the antibody is a humanized antibody.

[0428] Implementation Scheme 45. An isolated antibody from any of the foregoing implementation schemes, wherein the antibody is antagonistic to human CD73.

[0429] Implementation Scheme 46. The isolated antibody of Implementation Scheme 45, wherein the antibody deactivates, reduces or inhibits the activity of human CD73.

[0430] Implementation Scheme 47. The isolated antibody of Implementation Scheme 45 or 46, wherein the antibody weakens the ability of human CD73 to convert adenosine monophosphate to adenosine.

[0431] Implementation Scheme 48. An isolated antibody from any of the preceding implementation schemes, wherein the antibody is internalized upon binding to cells expressing human CD73.

[0432] Implementation Scheme 49. An isolated antibody from any one of Implementation Schemes 1-48, further comprising a TGFβ binding moiety.

[0433] Implementation scheme 50. The isolated antibody of implementation scheme 49, wherein the TGFβ binding portion is linked to the heavy chain variable region.

[0434] Implementation Scheme 51. The isolated antibody of Implementation Scheme 50, wherein the TGFβ binding moiety is linked to the heavy chain variable region via a peptide linker.

[0435] Implementation Scheme 52. An isolated antibody from any one of Implementation Schemes 30-48, further comprising a TGFβ binding moiety linked to the heavy chain constant region.

[0436] Implementation Scheme 53. The isolated antibody of Implementation Scheme 52, wherein the TGFβ binding moiety is linked to the C-terminal residue of the heavy chain constant region.

[0437] Implementation Scheme 54. The isolated antibody of Implementation Scheme 52 or 53, wherein the N-terminal residue of the TGFβ binding moiety is linked to the C-terminal residue of the heavy chain constant region via a peptide linker.

[0438] Implementation Scheme 55. The isolated antibody of Implementation Scheme 51 or 54, wherein the peptide linker comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 103-108.

[0439] Implementation Scheme 56. An isolated antibody from any one of Implementation Schemes 49-55, wherein the TGFβ binding portion specifically binds to human TGFβ.

[0440] Implementation Scheme 57. An isolated antibody of any one of Implementation Schemes 49-56, wherein the TGFβ binding portion comprises the extracellular domain of the human TGFβ receptor.

[0441] Implementation scheme 58. The isolated antibody of implementation scheme 57, wherein the human TGFβ receptor is selected from the group consisting of human TGFβR1, TGFβR2 and TGFβR3.

[0442] Implementation scheme 59. The isolated antibody of implementation scheme 57 or 58, wherein the human TGFβ receptor is human TGFβR2.

[0443] Implementation Scheme 60. An isolated antibody of any one of Implementation Schemes 49-59, wherein the TGFβ binding portion comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0444] Implementation Scheme 61. An isolated antibody of any one of Implementation Schemes 49-60, wherein the TGFβ binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0445] Implementation Scheme 62. An isolated antibody of any one of Implementation Schemes 49-61, wherein the TGFβ binding portion comprises an amino acid sequence selected from the group consisting of SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0446] Implementation Scheme 63. An isolated antibody of any one of Implementation Schemes 49-62, wherein the TGFβ binding moiety consists of an amino acid sequence selected from the group consisting of: SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0447] Implementation Scheme 64. An isolated antibody of any one of Implementation Schemes 49-63, said antibody comprising a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 113-121, preferably SEQ ID NO: 114 or 115.

[0448] Implementation Scheme 65. An isolated antibody of any one of Implementation Schemes 49-64, wherein the antibody comprises a polypeptide comprising an amino acid sequence selected from SEQ ID NO:113-121 and a polypeptide comprising an amino acid sequence comprising SEQ ID NO:92 or 97.

[0449] Implementation Scheme 66. An isolated antibody of any one of Implementation Schemes 49-65, wherein the antibody comprises two polypeptides each comprising an amino acid sequence selected from SEQ ID NO:113-121 and two polypeptides each comprising an amino acid sequence of SEQ ID NO:92 or 97.

[0450] Implementation Scheme 67. An isolated antibody of any one of Implementation Schemes 49-66, wherein the antibody comprises two polypeptides each comprising the amino acid sequence of SEQ ID NO:115 and two polypeptides each comprising the amino acid sequence of SEQ ID NO:97.

[0451] Implementation Scheme 68. An isolated antibody that specifically binds to human CD73, said antibody comprising a TGFβ binding moiety.

[0452] Implementation Scheme 69. The isolated antibody of Implementation Scheme 68, wherein the antibody is antagonistic to human CD73.

[0453] Implementation Scheme 70. The isolated antibody of Implementation Scheme 69, wherein the antibody deactivates, reduces or inhibits the activity of human CD73.

[0454] Implementation Scheme 71. The isolated antibody of Implementation Scheme 69 or 70, wherein the antibody weakens the ability of human CD73 to convert adenosine monophosphate to adenosine.

[0455] Implementation Scheme 72. An isolated antibody from any one of Implementation Schemes 68-71, wherein the antibody is antagonistic to human CD73 when it binds to TGFβ.

[0456] Implementation Scheme 73. An isolated antibody from any one of Implementation Schemes 68-72, wherein the TGFβ binding portion specifically binds to human TGFβ.

[0457] Implementation Scheme 74. An isolated antibody of any one of Implementation Schemes 68-73, wherein the TGFβ binding portion comprises the extracellular domain of the human TGFβ receptor.

[0458] Implementation Scheme 75. An isolated antibody from any one of Implementation Schemes 68-74, wherein the human TGFβ receptor is selected from the group consisting of human TGFβR1, TGFβR2 and TGFβR3.

[0459] Implementation Scheme 76. The isolated antibody of Implementation Scheme 74 or 75, wherein the human TGFβ receptor is human TGFβR2.

[0460] Implementation Scheme 77. An isolated antibody of any one of Implementation Schemes 68-76, wherein the TGFβ binding portion comprises an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0461] Implementation Scheme 78. An isolated antibody of any one of Implementation Schemes 68-77, wherein the TGFβ binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a sequence selected from the group consisting of: SEQ ID NO: 109-112.

[0462] Implementation Scheme 79. An isolated antibody of any one of Implementation Schemes 68-78, wherein the TGFβ binding portion comprises an amino acid sequence selected from the group consisting of SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0463] Implementation Scheme 80. An isolated antibody of any one of Implementation Schemes 68-79, wherein the TGFβ-binding moiety comprises an amino acid sequence selected from the group consisting of: SEQ ID NO:109-112, preferably SEQ ID NO:111.

[0464] Implementation Scheme 81. An isolated antibody from any of Implementation Schemes 1-48, further comprising a VEGF-binding moiety.

[0465] Implementation Scheme 82. The isolated antibody of Implementation Scheme 81, wherein the VEGF binding portion is linked to the heavy chain variable region.

[0466] Implementation Scheme 83. The isolated antibody of Implementation Scheme 82, wherein the VEGF binding moiety is linked to the heavy chain variable region via a peptide linker.

[0467] Implementation Scheme 84. An isolated antibody of any one of Implementation Schemes 30-48, further comprising a VEGF binding moiety linked to the heavy chain constant region.

[0468] Implementation Scheme 85. The isolated antibody of Implementation Scheme 84, wherein the VEGF binding portion is linked to the C-terminal residue of the heavy chain constant region.

[0469] Implementation Scheme 86. The isolated antibody of Implementation Scheme 84 or 85, wherein the N-terminal residue of the VEGF binding moiety is linked to the C-terminal residue of the heavy chain constant region via a peptide linker.

[0470] Implementation Scheme 87. The isolated antibody of Implementation Scheme 83 or 86, wherein the peptide linker comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 103-108.

[0471] Implementation Scheme 88. An isolated antibody from any one of Implementation Schemes 81-87, wherein the VEGF binding portion specifically binds to human VEGF.

[0472] Implementation Scheme 89. An isolated antibody of any one of Implementation Schemes 81-88, wherein the VEGF binding portion comprises the extracellular domain of the human VEGF receptor.

[0473] Implementation scheme 90. An isolated antibody of any one of implementation schemes 81-89, wherein the human VEGF receptor is selected from the group consisting of human VEGFR1, VEGFR2 and VEGFR3.

[0474] Implementation scheme 91. The isolated antibody of implementation scheme 89 or 90, wherein the human VEGF receptor is human VEGFR1.

[0475] Implementation Scheme 92. An isolated antibody of any one of Implementation Schemes 81-91, wherein the VEGF binding portion comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the same amino acid sequence as the sequence shown in SEQ ID NO:122.

[0476] Implementation Scheme 93. An isolated antibody of any one of Implementation Schemes 81-92, wherein the VEGF binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO:122.

[0477] Implementation Scheme 94. An isolated antibody of any one of Implementation Schemes 81-93, wherein the VEGF binding portion comprises the sequence shown in SEQ ID NO:122.

[0478] Implementation Scheme 95. An isolated antibody of any one of Implementation Schemes 81-94, wherein the VEGF binding portion comprises the sequence shown in SEQ ID NO:122.

[0479] Implementation Scheme 96. An isolated antibody of any one of Implementation Schemes 81-92, said antibody comprising a polypeptide comprising an amino acid sequence selected from the group consisting of: SEQ ID NO:123-127.

[0480] Implementation Scheme 97. An isolated antibody of any one of Implementation Schemes 81-96, wherein the antibody comprises a polypeptide comprising an amino acid sequence selected from SEQ ID NO:123-127 and a polypeptide comprising an amino acid sequence selected from SEQ ID NO:92, 128 and 97.

[0481] Implementation Scheme 98. An isolated antibody of any one of Implementation Schemes 81-97, wherein the antibody comprises two polypeptides comprising an amino acid sequence selected from SEQ ID NO:123-127 and two polypeptides comprising an amino acid sequence selected from SEQ ID NO:92, 128 and 97.

[0482] Implementation scheme 99. An isolated antibody that specifically binds to human CD73, said antibody comprising a VEGF binding moiety.

[0483] Implementation scheme 100. The isolated antibody of implementation scheme 99, wherein the antibody is antagonistic to human CD73.

[0484] Implementation Scheme 101. The isolated antibody of Implementation Scheme 100, wherein the antibody deactivates, reduces or inhibits the activity of human CD73.

[0485] Implementation Scheme 102. The isolated antibody of Implementation Scheme 100, wherein the antibody weakens the ability of human CD73 to convert adenosine monophosphate into adenosine.

[0486] Implementation scheme 103. An isolated antibody of any one of implementation schemes 99-102, wherein the antibody is antagonistic to human CD73 when it binds to VEGF.

[0487] Implementation scheme 104. An isolated antibody from any one of implementation schemes 99-103, wherein the VEGF binding portion specifically binds to human VEGF.

[0488] Implementation Scheme 105. An isolated antibody of any one of Implementation Schemes 99-104, wherein the VEGF binding portion comprises the extracellular domain of the human VEGF receptor.

[0489] Implementation scheme 106. An isolated antibody of any one of implementation schemes 99-105, wherein the human VEGF receptor is selected from the group consisting of human VEGFR1, VEGFR2 and VEGFR3.

[0490] Implementation scheme 107. The isolated antibody of implementation scheme 105 or 106, wherein the human VEGF receptor is human VEGFR1.

[0491] Implementation Scheme 108. An isolated antibody of any one of Implementation Schemes 99-107, wherein the VEGF binding portion comprises at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the amino acid sequence identical to the sequence shown in SEQ ID NO:122.

[0492] Implementation Scheme 109. An isolated antibody of any one of Implementation Schemes 99-108, wherein the VEGF binding portion consists of an amino acid sequence that is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence shown in SEQ ID NO:122.

[0493] Implementation Scheme 110. An isolated antibody of any one of Implementation Schemes 99-109, wherein the VEGF binding portion comprises the amino acid sequence shown in SEQ ID NO:122.

[0494] Implementation Scheme 111. An isolated antibody of any one of Implementation Schemes 99-110, wherein the VEGF binding portion consists of the amino acid sequence shown in SEQ ID NO:122.

[0495] Implementation Scheme 112. The isolated antibody of any of the foregoing implementation schemes further comprises a conjugated cytotoxic agent, cell growth inhibitor, toxin, radionuclide, or detectable marker.

[0496] Implementation Scheme 113. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to an epitope located in a region of human CD73, the amino acid sequence of which consists of the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0497] Implementation Scheme 114. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to at least one residue in the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0498] Implementation Scheme 115. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to one or more amino acid residues selected from the group consisting of human CD73: Y158, Y161, P165 or D168 according to the amino acid sequence number of SEQ ID NO:99.

[0499] Implementation Scheme 116. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 133 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0500] Implementation Scheme 117. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 134 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0501] Implementation Scheme 118. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to amino acid residue Y158 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0502] Implementation Scheme 119. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 59 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0503] Implementation Scheme 120. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to amino acid residue Y161 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0504] Implementation Scheme 121. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 60 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0505] Implementation Scheme 122. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to amino acid residue P165 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0506] Implementation Scheme 123. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 139 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0507] Implementation Scheme 124. An isolated antibody from any of the preceding implementation schemes, wherein the antibody binds to amino acid residue D168 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0508] Implementation Scheme 125. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 140 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0509] Implementation Scheme 126. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 135 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0510] Implementation Scheme 127. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 136 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0511] Implementation Scheme 128. An isolated antibody from any of the preceding implementation schemes, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 137 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0512] Implementation Scheme 129. An isolated antibody from any of the preceding embodiments, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 138 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0513] Implementation Scheme 130. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to an epitope located in a region of human CD73, the amino acid sequence of said region consisting of the amino acid sequences of SEQ ID NO: 90 and / or 91.

[0514] Implementation Scheme 131. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to at least one residue in the amino acid sequence of SEQ ID NO: 90 and / or 91.

[0515] Implementation Scheme 132. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to one or more amino acid residues of human CD73 selected from the group consisting of: Y158, Y161, P165 or D168 according to the amino acid sequence number of SEQ ID NO:99.

[0516] Implementation Scheme 133. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 133 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0517] Implementation Scheme 134. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 134 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0518] Implementation scheme 135. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to amino acid residue Y158 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0519] Implementation Scheme 136. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 59 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0520] Implementation scheme 137. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to amino acid residue Y161 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0521] Implementation Scheme 138. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 60 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0522] Implementation scheme 139. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to amino acid residue P165 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0523] Implementation Scheme 140. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 139 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0524] Implementation Scheme 141. An isolated antibody that specifically binds to human CD73, wherein the antibody binds to amino acid residue D168 of human CD73 according to the amino acid sequence number of SEQ ID NO: 99.

[0525] Implementation Scheme 142. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 140 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0526] Implementation Scheme 143. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 135 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0527] Implementation Scheme 144. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 136 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0528] Implementation Scheme 145. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 137 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0529] Implementation Scheme 146. An isolated antibody that specifically binds to human CD73, wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 142 is significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141, and wherein the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 138 is not significantly reduced relative to the binding affinity of the antibody to a protein comprising the amino acid sequence of SEQ ID NO: 141.

[0530] Implementation scheme 147. A compound or molecule comprising a CD73 binding moiety and a TGFβ binding moiety.

[0531] Implementation scheme 148. An isolated polynucleotide encoding a polypeptide of an antibody of any one of implementation schemes 1-146.

[0532] Implementation Scheme 149. The polynucleotide of Implementation Scheme 148, wherein the polynucleotide comprises cDNA or mRNA.

[0533] Implementation scheme 150. A vector comprising the polynucleotides of implementation scheme 148 or 149.

[0534] Implementation scheme 151. The vector of implementation scheme 150, wherein the vector is a plasmid vector or a viral vector.

[0535] Implementation scheme 152. A recombinant host cell containing the polynucleotide of implementation scheme 148 or 149 or the vector of implementation scheme 150 or 151.

[0536] Implementation Scheme 153. Lipid nanoparticles comprising the polynucleotide of Implementation Scheme 148 or 149 or the carrier of Implementation Scheme 150 or 151.

[0537] Implementation Scheme 154. A pharmaceutical composition comprising an antibody, polynucleotide, carrier or lipid nanoparticle, and a pharmaceutically acceptable carrier or excipient of any one of Implementation Schemes 1-146 or 148-153.

[0538] Implementation Scheme 155. A method for generating antibodies, the method comprising culturing a host cell of Implementation Scheme 152 under suitable conditions to express the polynucleotide and generate the antibody.

[0539] Implementation Scheme 156. A method for treating cancer in a subject, the method comprising administering to the subject an effective amount of an antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153, or 154.

[0540] Implementation Scheme 157. A method for inhibiting cancer metastasis in a subject, the method comprising administering to the subject an effective amount of an antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153, or 154.

[0541] Implementation scheme 158. The method of implementation scheme 156 or 157, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered intravenously, intratumorally, subcutaneously, intradermally, intramuscularly, intrabladderally, intracranially, intracavitarily or intraventricularly.

[0542] Implementation scheme 159. The method of implementation scheme 156 or 157, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered intratumorally.

[0543] Implementation scheme 160. The method of any one of implementation schemes 156-159, further comprising administering one or more additional therapeutic agents to the subject.

[0544] Implementation Scheme 161. The method of any one of Implementation Schemes 156-160, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition and the one or more additional therapeutic agents are co-administered according to the same schedule (e.g., co-administered at the same time intervals).

[0545] Implementation Scheme 162. The method of any one of Implementation Schemes 156-160, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition and one or more additional therapeutic agents are co-administered according to different schedules (e.g., co-administered at different time intervals).

[0546] Implementation Scheme 163. The method of any one of Implementation Schemes 156-162, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered in combination with one or more additional therapeutic agents, said one or more additional therapeutic agents comprising an activator or agonist of an FMS-associated tyrosine kinase 3 (FLT3; CD135) receptor, a Toll-like receptor (TLR) or an interferon gene stimulator (STING) receptor.

[0547] Implementation Scheme 164. The method of Implementation Scheme 163, wherein the TLR agonist or activator is selected from the group consisting of: TLR2 agonists, TLR3 agonists, TLR7 agonists, TLR8 agonists and TLR9 agonists.

[0548] Implementation Scheme 165. The method of Implementation Scheme 163, wherein the STING receptor agonist or activator is selected from the group consisting of: ADU-S100 (MIW-815), SB-11285, MK-1454, SR-8291, AdVCA0848, GSK-532, SYN-STING, MSA-1, SR-8291, 5,6-dimethylxanthenone-4-acetic acid (DMXAA), cyclic GAMP (cGAMP), and cyclic diAMP.

[0549] Implementation Scheme 166. The method of any one of Implementation Schemes 156-165, wherein the antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition is co-administered with one or more additional therapeutic agents, said one or more additional therapeutic agents comprising inhibitors or antagonists of: non-receptor protein tyrosine phosphatase 11 (PTPN11 or SHP2), myeloid leukemia sequence 1 (MCL1) apoptosis regulator; mitogen-activated protein kinase kinase 1 (MAP4K1) (also known as hematopoietic progenitor cell kinase 1 (HPK1)); diacylglycerol kinase alpha (DGKA, DAGK, DAGK1, or DGK-alpha); extracellular 5'-nucleotidase (NT5E or CD73); transforming growth factor beta 1 (TGFB1 or TGFβ); heme oxygenase 1 (HMOX1, HO-1, or HO1); vascular endothelial growth factor A (VEGFA or VEGF); erb-b2 receptor tyrosine kinase 2 (ERBB2). HER2, HER2 / neu, or CD340; epidermal growth factor receptor (EGFR, ERBB, ERBB1, or HER1); ALK receptor tyrosine kinase (ALK, CD246); poly(ADP-ribose) polymerase 1 (PARP1 or PARP); cyclin-dependent kinase 4 (CDK4); cyclin-dependent kinase 6 (CDK6); CC motif chemokine receptor 8 (CCR8, CDw198); CD274 molecule (CD274, PDL1, or PD-L1); programmed cell death 1 (PDCD1, PD1, or PD-1); and / or cytotoxic T lymphocyte-associated protein 4 (CTLA4, CTLA-4, CD152).

[0550] Implementation Scheme 167. The method of Implementation Scheme 166, wherein the inhibitor comprises an antigen-binding molecule, an antibody, or an antigen-binding fragment thereof.

[0551] Implementation scheme 168. The method of implementation scheme 166, wherein the inhibitor comprises small organic molecules.

[0552] Implementation scheme 169. The method of implementation scheme 166, wherein the MCL1 inhibitor is selected from the following group: AMG-176, AMG-397, S-64315, AZD-5991, 483-LM, A1210477, UMI-77 and JKY-5-037.

[0553] Implementation scheme 170. The method of implementation scheme 166, wherein the inhibitor of PTPN11 or SHP2 is selected from the group consisting of: TNO155 (SHP-099), RMC-4550, JAB-3068 and RMC-4630.

[0554] Implementation Scheme 171. The method of Implementation Scheme 160, wherein the additional therapeutic agent is a chemotherapeutic agent, an anti-biological agent, a radiotherapy agent, or a checkpoint targeting agent.

[0555] Implementation Scheme 172. The method of Implementation Scheme 160, wherein the one or more anti-biological agents or chemotherapeutic agents are selected from the group consisting of: nucleoside analogs (e.g., 5-fluorouracil, gemcitabine, cytarabine), taxanes (e.g., paclitaxel, nab-paclitaxel, docetaxel, cabazitaxel), platinum complexes (cisplatin, carboplatin, oxaliplatin, nedaplatin, triplatin).tetranitrate, phenanthriplatin, picoplatin, satraplatin, dicycloplatin, eptaplatin, lobaplatin, miriplatin, dihydrofolate reductase (DHFR) inhibitors (e.g., methotrexate, trimetrexate, pemetrexed), topoisomerase inhibitors (e.g., doxorubicin, daunorubicin, dactinomycin, eniposide, epirubicin, etoposide, idarubicin, irinotecan, mitoxantrone, pixantrone, sobuzoxane, topotecan otecan), irinotecan, MM-398 (irinotecan liposome), vosaroxin and GPX-150, aldoxorubicin, AR-67, growlertinib, AST-2818, avitinib (ACEA-0010), irofulven (MGI-114), alkylating agents (e.g., nitrogen mustard, cyclophosphamide, chlormethine, urethane) Uramustine or uramustine, melphalan, chlorambucil, ifosfamide, bendamustine, temozolomide, carmustine, nitrosoureas (e.g., carmustine, lomustine, streptozocin), alkyl sulfonates (e.g., busulfan), and mixtures thereof.

[0556] Implementation Scheme 173. The method of Implementation Scheme 160, wherein the checkpoint targeting agent is selected from the group consisting of: antagonist anti-PD-1 antibody, antagonist anti-PD-L1 antibody, antagonist anti-PD-L2 antibody, antagonist anti-CTLA-4 antibody, antagonist anti-TIM-3 antibody, antagonist anti-LAG-3 antibody, antagonist anti-CEACAM1 antibody, agonist anti-GITR antibody, antagonist anti-TIGIT antibody, antagonist anti-VISTA antibody, agonist anti-CD137 antibody, and agonist anti-OX40 antibody.

[0557] Implementation Scheme 174. The method of Implementation Scheme 173, wherein the additional therapeutic agent is an anti-PD-1 antibody, optionally wherein the anti-PD-1 antibody is pembrolizumab or nivolumab.

[0558] Implementation Scheme 175. The method of Implementation Scheme 160, wherein the additional therapeutic agent is an inhibitor of indoleamine-2,3-dioxygenase (IDO).

[0559] Implementation Scheme 176. The method of Implementation Scheme 175, wherein the inhibitor is selected from the group consisting of: epacadostat, F001287, indoximod and NLG919.

[0560] Implementation scheme 177. The method of implementation scheme 176, wherein the inhibitor is edodestat.

[0561] Implementation scheme 178. The method of implementation scheme 160, wherein the additional therapeutic agent is a vaccine.

[0562] Implementation Scheme 179. The method of Implementation Scheme 178, wherein the vaccine comprises a heat shock peptide complex (HSPPC), the HSPPC comprising a heat shock protein complexed with an antigenic peptide.

[0563] Implementation scheme 180. The method of implementation scheme 179, wherein the heat shock protein is hsc70 and is complexed with a tumor-associated antigenic peptide.

[0564] Implementation Scheme 181. The method of Implementation Scheme 179, wherein the heat shock protein is gp96 and is complexed with a tumor-associated antigenic peptide, wherein the HSPPC is derived from a tumor obtained from the subject.

[0565] Implementation Scheme 182. The method of Implementation Scheme 160, wherein the additional therapeutic agent is etoposide or doxorubicin.

[0566] Implementation Scheme 183. The method of any one of Implementation Schemes 156-182, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered in combination with the FOLFOX regimen, the FOLFOXIRI regimen or the FOLFIRINOX regimen.

[0567] Implementation scheme 184. The methods of implementation schemes 156-183, wherein the antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition is administered in combination with immunotherapy, immunostimulation therapy, cell therapy or gene therapy.

[0568] Implementation scheme 185. The method of any one of implementation schemes 156-184, wherein the cancer is selected from the group consisting of solid tumors, hematologic cancers and metastatic lesions.

[0569] Implementation scheme 186. The method of implementation scheme 185, wherein the solid tumor is selected from the group consisting of sarcoma, fibroblastic sarcoma, carcinoma and adenocarcinoma.

[0570] Implementation scheme 187. The method of implementation scheme 185, wherein the hematologic cancer is selected from the group consisting of leukemia, lymphoma and myeloma.

[0571] Implementation Scheme 188. The method of any one of Implementation Schemes 156-184, wherein the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloid monocytic leukemia, etc. Nucleated cell leukemia (CMML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma.

[0572] Implementation Scheme 189. The method of any one of Implementation Schemes 156-184, wherein the cancer is selected from the group consisting of: epithelial tumors (e.g., carcinoma, squamous cell carcinoma, basal cell carcinoma, squamous intraepithelial neoplasm), adenomatous tumors (e.g., adenocarcinoma, adenoma, adenomyoma), stromal or soft tissue tumors (e.g., sarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, fibrosarcoma, dermatofibrosarcoma, neurofibrosarcoma, fibrous histiocytoma, angiosarcoma, angiomyosarcoma, leiomyosarcoma, chondroma, chondrosarcoma, alveolar soft sarcoma, epithelioid angioendothelioma, Spitz tumor, synovial sarcoma), and lymphoma.

[0573] Implementation Scheme 190. The method of any one of Implementation Schemes 156-184, wherein the cancer is a solid tumor in or originating from a tissue or organ selected from the group consisting of: bone (e.g., ameloblastoma, aneurysmal bone cyst, angiosarcoma, chondroblastoma, chondroma, chondromycinoma, chondrosarcoma, chordoma, dedifferentiated chondrosarcoma, enchondroma, epithelioid hemangioendothelioma, fibrous dysplasia of bone, giant cell tumor of bone, hemangioma and related lesions, osteoblastoma, osteochondroma, osteosarcoma, osteoid osteoma, osteoma, periosteal chondroma, desmoidoma, Ewing sarcoma); lips and oral cavity (e.g., odontogenic ameloblastoma, oral leukoplakia, oral squamous cell carcinoma, primary ameloblastoma, etc.). Primary oral mucosal melanoma; salivary glands (e.g., pleomorphic salivary gland adenoma, salivary gland adenoid cystic carcinoma, salivary gland mucoepidermoid carcinoma, salivary gland Warthin tumor); esophagus (e.g., Barrett's esophagus, dysplasia, and adenocarcinoma); gastrointestinal tract, including the stomach (e.g., gastric adenocarcinoma, primary gastric lymphoma, gastrointestinal stromal tumor (GIST), metastatic deposits, gastric carcinoid, gastric sarcoma, neuroendocrine carcinoma, primary squamous cell carcinoma of the stomach, gastric adenoacanthoma), intestine and smooth muscle (e.g., intravenous leiomyomatosis), colon (e.g., colorectal adenocarcinoma), rectum, anus; pancreas (e.g., serous neoplasms, including microcystic or macrocystic serous cystadenomas, solid serous cystadenomas, Von Hippel-Landau (VHL) related serous cystic neoplasms, serous cystadenocarcinoma, mucinous cystic neoplasms (MCN), intraductal papillary mucinous neoplasms (IPMN), intraductal eosinophilic papillary neoplasms (IOPN), intraductal tubular neoplasms, cystic acinar neoplasms including acinar cell cystadenoma, acinar cell cystadenocarcinoma, pancreatic adenocarcinoma, invasive pancreatic ductal adenocarcinoma including tubular adenocarcinoma, adenosquamous carcinoma, colloid carcinoma, medullary carcinoma, hepatoid carcinoma, signet ring cell carcinoma, undifferentiated carcinoma, undifferentiated carcinoma with osteoclast-like giant cells, acinar cell carcinoma, neuroendocrine neoplasms Biological organisms, including neuroendocrine microadenomas, neuroendocrine tumors (NETs), neuroendocrine carcinomas (NECs), including small cell or large cell NECs, insulinomas, gastrinomas, glucagonomas, serotonin-producing NETs, ​​somatostatinomas, angiotensinomas, solid pseudopapillary neoplasms (SPNs), pancreatoblastomas; gallbladder (e.g., gallbladder cancer and extrahepatic bile duct cancer, intrahepatic bile duct cancer); neuroendocrine (e.g., adrenocortical carcinoma, carcinoid tumors, pheochromocytomas, pituitary adenomas); thyroid (e.g., anaplastic (undifferentiated) carcinoma, medullary carcinoma, eosinophilic tumors, papillary carcinoma, adenocarcinoma);Liver (e.g., adenoma, combined hepatocellular and cholangiocarcinoma, fibrolamellar carcinoma, hepatoblastoma, hepatocellular carcinoma, mesenchymal, nested stromal epithelial tumors, undifferentiated carcinoma, hepatocellular carcinoma, intrahepatic cholangiocarcinoma, bile duct cystadenocarcinoma, epithelioid hemangioendothelioma, angiosarcoma, embryonal sarcoma, rhabdomyosarcoma, solitary fibroma, teratoma, yolk sac tumor, carcinosarcoma, rhabdomyosarcoma); kidney (e.g., ALK rearranged renal cell carcinoma, chromophobe renal cell carcinoma, clear cell renal cell carcinoma, clear cell sarcoma, metanephrotic adenoma, metanephrotic fibroma, mucinous tubular and spindle cell carcinoma, nephrotic tumor, nephroblastoma). Wilms' tumor, papillary adenoma, papillary renal cell carcinoma, renal eosinophilic cell carcinoma, renal cell carcinoma, succinate dehydrogenase-deficient renal cell carcinoma, collecting duct carcinoma); breast (e.g., invasive ductal carcinoma, including but not limited to acinic cell carcinoma, adenoid cystic carcinoma, apocrine gland carcinoma, cribriform carcinoma, glycogen-rich / clear cell carcinoma, inflammatory carcinoma, lipid-rich carcinoma, medullary carcinoma, metaplastic carcinoma, micropapillary carcinoma, mucinous carcinoma, neuroendocrine carcinoma, eosinophilic carcinoma, papillary carcinoma, sebaceous gland carcinoma, secretory breast cancer, tubular carcinoma, lobular carcinoma, including but not limited to pleomorphic carcinoma, immature carcinoma, etc.). Cellular carcinomas, peritoneum (e.g., mesothelioma, primary peritoneal carcinoma); female sex organ tissues, including ovaries (e.g., choriocarcinoma, epithelial tumors, germ cell tumors, sex cord-stromal tumors), fallopian tubes (e.g., serous adenocarcinoma, mucinous adenocarcinoma, endometrioid adenocarcinoma, clear cell adenocarcinoma, transitional cell carcinoma, squamous cell carcinoma, undifferentiated carcinoma, Müllerian tumor, adenosarcoma, leiomyosarcoma, teratoma, germ cell tumors, choriocarcinoma, trophoblastic tumors), uterus (e.g., cervical cancer, endometrial polyps, endometrial hyperplasia, epithelial intraepithelial carcinoma (EIC), endometrial cancer). (e.g., endometrioid carcinoma, serous carcinoma, clear cell carcinoma, mucinous carcinoma, squamous cell carcinoma, transitional carcinoma, small cell carcinoma, undifferentiated carcinoma, mesenchymal neoplasms), leiomyomas (e.g., endometrial stromal nodules, leiomyosarcoma, endometrial stromal sarcoma (ESS), mesenchymal tumors), mixed epithelial and mesenchymal tumors (e.g., adenofibroma, carcinoma fibroma, adenosarcoma, carcinoma sarcoma (malignant mixed mesodermal sarcoma - MMMT), endometrial stromal tumors, malignant mixed Müllerian tumors of the endometrium, gestational trophoblastic tumors (partial hydatiform mole) Mole, complete hydatidiform mole, invasive hydatidiform mole, placental site tumors); vulva, vagina; male sex organs, including prostate, testes (e.g., germ cell tumors, seminoma), penis; bladder (e.g., squamous cell carcinoma, urothelial carcinoma, bladder urothelial carcinoma); brain (e.g., gliomas (e.g., astrocytomas, including non-invasive, low-grade, anaplastic, glioblastomas; oligodendrogliomas, ependymomas), meningiomas, gangliogliomas, Schwann cell tumors (schwannomas), craniopharyngiomas, chordomas, non-Hodgkin lymphomas, pituitary tumors);Eye (e.g., retinoma, retinoblastoma, ocular melanoma, retrouvian melanoma, iris hamartoma); head and neck (e.g., nasopharyngeal carcinoma, endolymphatic cystic tumor (ELST), epidermoid carcinoma, laryngeal carcinoma including squamous cell carcinoma (SCC) (e.g., glottic carcinoma, supraglottic carcinoma, subglottic carcinoma, transglottic carcinoma), carcinoma in situ, verrucous, spindle cell and basaloid SCC, undifferentiated carcinoma, laryngeal adenocarcinoma, adenoid cystic carcinoma, neuroendocrine carcinoma, laryngeal sarcoma), head and neck paraganglioma (e.g., carotid body, tympanic cavity, vagus nerve); thymus (e.g., thymoma); heart (e.g., cardiac myxoma); lung (e.g., small cell carcinoma (SCLC), non-small cell lung cancer (NSCLC), including squamous cell carcinoma (SCC), adenocarcinoma and large cell carcinoma, carcinoid (typical or atypical), carcinosarcoma, pulmonary blastoma, giant cell carcinoma). Spindle cell carcinoma, pleural pulmonary blastoma; lymphoma (e.g., lymphoma, including Hodgkin's lymphoma, non-Hodgkin's lymphoma, EBV-associated lymphoproliferative disorders, including B-cell lymphoma and T-cell lymphoma (e.g., Burkitt lymphoma, large B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, indolent B-cell lymphoma, low-grade B-cell lymphoma, fibrin-associated diffuse large cell lymphoma; primary exudative lymphoma; plasmablastic lymphoma; extranodal NK / T-cell lymphoma (nasal type); peripheral T-cell lymphoma, cutaneous T-cell lymphoma, angioimmunoblastic T-cell lymphoma; follicular T-cell lymphoma; systemic T-cell lymphoma, lymphangioleiomyomatosis);Central nervous system (CNS) tumors (e.g., gliomas, including astrocytic tumors such as pilocytic astrocytoma, pilocyxoid astrocytoma, subependymal giant cell astrocytoma, pleomorphic xanthoastrocytoma, diffuse astrocytoma, fibrous astrocytoma, and gemistocytic astrocytoma). astrocytoma, protoplasmic astrocytoma, anaplastic astrocytoma, glioblastoma (e.g., giant cell glioblastoma, glioma, glioblastoma multiforme, and gliomatosis), oligodendroglioma (e.g., oligodendroglioma, anaplastic oligodendroglioma), oligodendroastrocytic tumor (e.g., oligodendroglioma, anaplastic oligodendroglioma), ependymal tumor (e.g., subependymoma, myxopapillary ependymoma, ependymoma (e.g., cellular, papillary, clear cell, elongated cell), anaplastic ependymoma), optic glioma and non-glial tumor (e.g., choroid plexus tumors, neuronal and mixed neuronal glial tumors, pineal region tumors, embryonal tumors). Tumors, including medulloblastomas, meningeal tumors, primary CNS lymphomas, germ cell tumors, pituitary adenomas, cranial nerve and paravertebral nerve tumors, stellate region tumors, neurofibromas, meningiomas, peripheral nerve sheath tumors, peripheral neuroblastomas (including but not limited to neuroblastomas, ganglioneuromas, ganglioneuromas), trisomy 19 ependymomas; neuroendocrine tissues (e.g., paraganglionic systems, including adrenal medulla (pheochromocytoma) and extra-adrenal paragangliomas (extra-adrenal paragangliomas); skin (e.g., clear cell hidradenoma, benign fibrous histiocytoma of the skin, cylindrica, hidradenoma, melanoma (including cutaneous melanoma, mucosal melanoma), pilomatoma, Spitz tumors);and soft tissue (e.g., aggressive angiomyxoma, alveolar rhabdomyosarcoma, alveolar soft sarcoma, angiofibroma, hemangioma-like fibrous histiocytoma, synovial sarcoma, biphasic synovial sarcoma, clear cell sarcoma, dermatofibrosarcoma protuberans, desmoid fibromatosis, small round cell tumor, desmoplastic small round cell tumor, elastic fibroma, embryonal rhabdomyosarcoma, Ewing's tumor / primary neuroectodermal tumor (PNET), bone) Extrailiac myxoid chondrosarcoma, extraskeletal osteosarcoma, paravertebral sarcoma, inflammatory myofibroblastic tumor, lipoblastoma, lipoma, chondroid lipoma, liposarcoma / malignant lipomatous tumor, liposarcoma, myxoid liposarcoma, fibromyxoid sarcoma, lymphangioleiomyosarcoma, malignant myoepithelioma, soft tissue malignant melanoma, myoepithelial carcinoma, myoepithelioma, myxoid inflammatory fibroblastic sarcoma, undifferentiated sarcoma, peridermoid cell tumor, rhabdomyosarcoma, non-rhabdomyosarcoma soft tissue sarcoma, soft tissue leiomyosarcoma, undifferentiated sarcoma, well-differentiated liposarcoma.

[0574] Implementation Scheme 191. The method of any one of Implementation Schemes 156-184, wherein the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal cancer, and endometrial cancer.

[0575] Implementation scheme 192. An isolated antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any one of implementation schemes 1-146, 148-151, 153 or 154, for the treatment of cancer.

[0576] Implementation scheme 193. Implementation scheme 192 uses isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions, wherein the cancer is selected from the group consisting of solid tumors, hematologic cancers and metastatic lesions.

[0577] Implementation scheme 194. Implementation scheme 192 uses isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions, wherein the solid tumor is selected from the group consisting of sarcoma, fibroblastic sarcoma, carcinoma and adenocarcinoma.

[0578] Implementation scheme 195. Implementation scheme 192 uses isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions, wherein the hematologic cancer is selected from the group consisting of leukemia, lymphoma and myeloma.

[0579] Implementation Scheme 196. The isolated antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition used in any of Implementation Schemes 192-195, wherein the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CLL), and chronic myeloid leukemia (CLL). Blood disorders (CML), chronic myeloid monocytic leukemia (CMML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma.

[0580] Implementation Scheme 197. An isolated antibody, polynucleotide, carrier, lipid nanoparticle, or pharmaceutical composition of any one of Implementation Schemes 192-195, wherein the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal cancer, and endometrial cancer.

[0581] Implementation Scheme 198. Use of isolated antibodies, polynucleotides, carriers, lipid nanoparticles or pharmaceutical compositions of any one of Implementation Schemes 1-146, 148-151, 153 or 154 in the preparation of a medicament for the treatment of cancer.

[0582] Implementation scheme 199. Use of implementation scheme 198, wherein the cancer is selected from the group consisting of solid tumors, hematologic cancers and metastatic lesions.

[0583] Implementation scheme 200. Use of implementation scheme 199, wherein the solid tumor is selected from the group consisting of sarcoma, fibroblastic sarcoma, carcinoma and adenocarcinoma.

[0584] Implementation scheme 201. Use of implementation scheme 199, wherein the hematologic cancer is selected from the group consisting of leukemia, lymphoma and myeloma.

[0585] Implementation Scheme 202. Use of any one of Implementation Schemes 198-201, wherein the cancer is selected from the group consisting of: lung cancer, melanoma, kidney cancer, liver cancer, myeloma, prostate cancer, breast cancer, ovarian cancer, colorectal cancer, pancreatic cancer, head and neck cancer, anal cancer, gastroesophageal cancer, mesothelioma, nasopharyngeal carcinoma, thyroid cancer, cervical cancer, epithelial cancer, peritoneal cancer, lymphoproliferative disorders, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloid monocytic leukemia, etc. Nucleated cell leukemia (CMML), hairy cell leukemia, B-cell lymphoma, diffuse large B-cell lymphoma (DLBCL), activated B-cell-like (ABC) diffuse large B-cell lymphoma, germinal center B-cell (GCB) diffuse large B-cell lymphoma, mantle cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, relapsed non-Hodgkin lymphoma, refractory non-Hodgkin lymphoma, relapsed follicular non-Hodgkin lymphoma, Burkitt lymphoma, small lymphocytic lymphoma, follicular lymphoma, lymphoplasmacytic lymphoma, and extranodal marginal zone lymphoma.

[0586] Implementation Scheme 203. Use of any one of Implementation Schemes 198-201, wherein the cancer is selected from the group consisting of: melanoma, gastric cancer, triple-negative breast cancer (TNBC), non-small cell lung cancer (NSCLC), rectal adenocarcinoma, colorectal cancer, renal cell carcinoma, ovarian cancer, prostate cancer, oral squamous cell carcinoma (SCC), head and neck squamous cell carcinoma (HNSCC), urothelial bladder cancer, glioblastoma (GBM), meningioma, adrenal cancer, and endometrial cancer.

[0587] Implementation Scheme 204. An isolated antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153 or 154, used in the method of any one of Implementation Schemes 156-191.

[0588] Implementation Scheme 205. Use of the isolated antibody, polynucleotide, carrier, lipid nanoparticle or pharmaceutical composition of any one of Implementation Schemes 1-146, 148-151, 153 or 154 in the method of any one of Implementation Schemes 156-191. Brief description of the attached figures

[0590] Figure 1 This is a set of graphs showing the binding of anti-CD73 antibodies (BA013, BA014, BA015, or a reference antibody (“RA001”)) or allotype control antibodies to Chinese hamster ovary (CHO) cells ectopically expressing mouse, cynomolgus monkey, or human CD73 (from left to right in the figure). Median fluorescence intensity (MFI) values ​​are plotted against antibody concentrations.

[0591] Figure 2A-2B This is a set of charts showing anti-CD73 antibodies (BA013, BA014, BA015, or RA001) or isotype controls ( Figure 2A Or anti-CD73 antibody BA015 or isotype control ( Figure 2B The degree of inhibition of CD73 enzyme activity. Jurkat cells ( Figure 2A ) or CHO cells ( Figure 2B The percentage of CD73 activity ectopically expressed on the surface of the antibody was plotted as the level of luminescence observed in the absence of the antibody (measured in relative light units (RLU)).

[0592] Figures 3A-3C It is a set of charts and images showing the internalization of anti-CD73 antibodies and antibody-mediated internalization of CD73. Figure 3A The results of a cytotoxicity assay are shown, which measures antibody internalization by incubating CD73-expressing cells with a test antibody (BA015, RA001, or a second reference antibody (“RA002”)), an allotype control, or no antibody, and a secondary antibody conjugated with cytotoxic monomethylauristatin E (“MMAE”). The level of antibody internalization is plotted as the viability of CD73-expressing cells relative to antibody concentration (normalized relative to the viability of control cells (“NT”) not treated with any primary or secondary antibody). Figure 3B A series of representative images from the CD73 internalization assay are shown, in which expression -CD73 fusion protein in cells after staining the cells with the fusion protein Alexa Fluor TM After incubation with 488 ligand and treatment with anti-CD73 or control antibody, the samples were detected by confocal fluorescence microscopy. Figure 3C It shows the intracellular data from four images. -Quantitative charts of CD73, the four images over a 6-hour period, each condition per hour in the FITC channel (to detect) Alexa Fluor TM It was obtained from 488).

[0593] Figure 4A-4P This is a set of graphs showing the binding of isotope controls, RA001, RA002, BA015, and humanized anti-CD73 antibodies BA019-BA030 to CHO cells ectopically expressing cynomolgus monkey CD73, as measured by flow cytometry. MFI values ​​are plotted against antibody concentrations to display binding curves.

[0594] Figures 5A-5P This is a set of graphs showing the binding of isotope controls, RA001, RA002, BA015, and humanized anti-CD73 antibodies BA019-BA030 to CHO cells ectopically expressing human CD73, as measured by flow cytometry. A negative control without any antibodies (referred to as "antibody-free") was used. Figure 5A The comparison was performed. MFI values ​​were plotted against antibody concentration to display the binding curve.

[0595] Figures 6A-6P This is a set of graphs showing the degree of inhibition of CD73 enzyme activity by isotope control, RA001, RA002, BA015, and humanized anti-CD73 antibody BA019-BA030. The percentage of CD73 activity ectopically expressed on the surface of Jurkat cells is plotted as a luminescence level (measured in RLU) relative to the level in the absence of antibody.

[0596] Figures 7A-7P This is a set of graphs showing the degree of inhibition of CD73 enzyme activity by isotope control, RA001, RA002, BA015, and humanized anti-CD73 antibody BA019-BA030. The percentage of activity of immobilized recombinant CD73 is plotted as luminescence levels (measured in RLU) relative to the level in the absence of antibody.

[0597] Figure 8A-8M This is a set of graphs showing the results of a cytotoxicity assay that measures antibody internalization by incubating CD73-expressing cells with antibody-free, isotope-controlled, BA015 or humanized anti-CD73 antibody, and a secondary antibody conjugated with MMAE. The level of antibody internalization is plotted as the viability of CD73-expressing cells against antibody concentrations (normalized relative to the viability of control cells (“NT”) that were not treated with any primary or secondary antibody).

[0598] Figures 9A-9E This is a set of charts showing the ability of anti-CD73 antibody (BA012) or anti-CD73-TGFβ trap fusion protein (BA035) rather than isotype controls (isotype antibody controls and fusion protein controls containing such isotype antibodies and TGFβ traps) to bind to CD73 and inhibit CD73 enzyme activity. Figure 9A and 9B It shows that in the absence of ( Figure 9A ) or exist ( Figure 9B In the presence of TGFβ1, BA012 or BA035, rather than the allotype control, bind to CHO cells ectopically expressing mouse CD73. The binding level, as measured by flow cytometry, is plotted as an absolute MFI value. Figure 9A) or plotted as a percentage of the MFI value relative to the MFI value for each test condition in the absence of TGFβ1 ( Figure 9B ). Figure 9C-9E It shows that in the absence of ( Figure 9C and 9D ) or exist ( Figure 9E In the presence of TGFβ1, BA012 or BA035, rather than the allotype control, inhibited CD73 enzyme activity. The percentage of CD73 activity ectopically expressed on the surface of CHO cells was plotted as a luminescence level (measured by RLU) relative to the level in the absence of antibody.

[0599] Figure 10A and 10B This is a set of charts showing the antagonism of TGFβ-induced signal transduction by BA012 or BA035, rather than the isotype control. Figure 10A In this study, the relative luminescence levels associated with TGFβ1-induced luciferase expression in HEK293-SMAD reporter cells were plotted as percentage values ​​relative to the luminescence levels in the absence of antibodies. Figure 10B In this study, the expression level of red fluorescent protein (RFP), which is associated with TGFβ1-induced vimentin expression in A549-Vimentin-RFP cells, was plotted as the absolute fluorescence value minus the background fluorescence level in the absence of antibody or TGFβ1.

[0600] Figure 11A and 11B This is a set of charts showing the inhibition of tumor growth and metastasis by anti-CD73 antibodies (isotope control, BA012, and a reference antibody containing the variable domain of RA001 and the constant domain of mouse IgG2a-N297A, named RA001-IgG2a-N297A). Figure 11A The average calculated tumor volume is shown for 8–10 mice / groups that were subcutaneously implanted with CT26 tumor cells and treated with the indicated antibody. Figure 11B The number of tumor nodules in the lungs of each mouse after intravenous implantation of EMT6 tumor cells and treatment with BA012 or an isotype control antibody is shown.

[0601] Figure 12A and 12B It is a set of charts showing the inhibition of tumor growth by BA012, BA035 or the isotype control. Figure 12A The average calculated tumor volume is shown for 5–7 mice / groups that were subcutaneously implanted with TC1 tumor cells and treated with the indicated antibody. Figure 12B The calculated tumor volume is shown for 8–10 mice / groups subcutaneously implanted with EMT6 tumor cells and treated with the indicated antibody.

[0602] Figure 13A and13B This is a set of charts showing, as measured by flow cytometry, the effects of anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody or isotype TGFβ trap on human ( Figure 13A ) or crab-eating macaque ( Figure 13B The binding of CD8+ T cells was isolated from PBMCs. The median MFI value was plotted against the antibody concentration.

[0603] Figures 14A-14D This is a set of graphs showing the inhibitory effects of anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody, or isotype TGFβ trap on recombinant human CD73 enzyme activity in biochemical assays. Results from two experiments are shown. Immobilized plate-bound CD73 ( Figure 14A and 14B ) or soluble CD73 ( Figure 14C and 14D The activity of the antibody was plotted as the luminescence level relative to the level in the absence of the antibody (measured in RLU).

[0604] Figures 15A-15D This is a set of graphs showing the inhibitory effects of anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody, or isotype TGFβ trap on recombinant cynomolgus monkey CD73 enzyme activity in biochemical assays. Results from two experiments are shown. Immobilized plate-bound CD73 ( Figure 15A and 15B ) or soluble CD73 ( Figure 15C and 15D The activity of the antibody was plotted as the luminescence level relative to the level in the absence of the antibody (measured in RLU).

[0605] Figure 16A and 16B This is a set of charts showing the degree of inhibition of CD73 enzyme activity by anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody, or isotype TGFβ trap. Two experiments ( Figure 16A and Figure 16B The percentage of CD73 activity ectopically expressed on the surface of Jurkat cells was plotted as a luminescence level (measured in RLU) relative to the level in the absence of antibody.

[0606] Figures 17A-17CThis is a set of graphs showing the antagonism of anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody, or isotype TGFβ trap on TGFβ-induced signaling in the HEK293 SMAD-luciferase reporter assay. Each TGFβ isotype was tested separately. TGFβ-induced SMAD signaling activity (corresponding to luciferase activity in HEK293-SMAD-luciferase cells) is plotted as luminescence levels (measured in RLU) relative to the level in the absence of antibody.

[0607] Figure 18A-18I This is a set of graphs showing the antagonism of anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody, or isotype TGFβ trap on TGFβ-induced signaling in the A549-Vimentin-RFP reporter cytogenetic assay. Each TGFβ isotype was tested separately. Results from three experiments are shown. Two experiments were performed at room temperature. Figures 18A-18C And 18D-18F). An experiment was conducted at 37°C ( Figure 18G-18I TGFβ-induced signaling activity (associated with RFP expression in A549-Vimentin-RFP cells) was plotted as the MFI of RFP levels relative to the absence of antibodies.

[0608] Figures 19A-19D This is a set of graphs showing the internalization of anti-CD73 antibodies and antibody-mediated CD73 internalization. The graphs show the results of a cytotoxicity assay that measures antibody internalization by incubating CD73-expressing cells with anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), an allotype control antibody, or no antibody (each case accompanied by a secondary antibody conjugated with the cytotoxic agent DM1). The level of antibody internalization was measured by cell death indicated by the loss of fluorescence intensity (measured in RLU). The data are plotted as RLU of CD73-expressing cells relative to antibody concentration. Data from two experiments are shown.

[0609] Figure 20A and 20B This is a set of charts showing the effects of anti-CD73 antibody (BA025), anti-CD73-TGFβ trap fusion protein (BA037), isotype control antibody, isotype TGFβ trap, and Fc activity on anti-CD73-activated Jurkat cells (containing an NFAT-luciferase reporter construct and expressing FcγRIIA(H131)). Figure 20A ) or Jurkat FcγRIIIA(V158)( Figure 20BThe ability of the FcγR receptor on the antibody was measured in the presence of CHO cells expressing CD73 in each case. FcγR receptor activation was plotted as the level of fluorescence (measured in RLU) relative to the level in the absence of the antibody.

[0610] Figure 21 This is a graph showing the inhibition of metastasis by BA035. The graph shows the number of tumor nodules in the lungs of each mouse that received intravenous EMT6 tumor cells and were treated with BA012, BA035, or an isotype control antibody.

[0611] Figure 22 This is a graph showing the inhibition of tumor growth by BA012, BA035, or the isotype control. The graph shows the average calculated tumor volume of 5–7 mice / groups implanted with LCC tumor cells and treated with the indicated antibody.

[0612] Figure 23 This is a graph showing the inhibition of tumor growth by BA012, BA035, or their isoforms. The graph shows the average calculated tumor volume of 8–10 mice / groups subcutaneously implanted with Detroit562 tumor cells and treated with the indicated antibodies.

[0613] Figures 24A-24D This is a set of graphs showing the inhibition of tumor growth by administration of BA012, BA035, or the isotype control alone or in combination with anti-PD-1 antibody or doxorubicin. The graphs show the calculated tumor volume of 6–9 mice / groups subcutaneously implanted with EMT6 tumor cells and treated with the antibodies shown. Figure 24A Data from mice that did not receive combination therapy are shown. Figure 24B Data from mice that received anti-PD-1 in addition to the antibody shown are presented. Figure 24C and 24D Data from mice that received doxorubicin in addition to the antibody shown are presented. Figure 24D The charts in the middle show and Figure 24C The charts contain the same data, except that the same type of control data has been removed and the x-axis has been redrawn to better separate the data from the different treatment groups.

[0614] Figure 25 This is a graph showing the inhibition of tumor growth by BA012, BA035, or isotype controls in combination with etoposide, or by etoposide alone. The graph shows the mean calculated tumor volume of 5–8 mice / groups subcutaneously implanted with CT26 tumor cells and treated with the indicated antibodies. Detailed Explanation

[0616] This disclosure provides specific binding to and antagonism of CD73 (e.g., human CD73, cynomolgus monkey CD73, or mouse CD73) to CD73 function, such as the enzymatic activity that promotes the conversion of AMP to adenosine. Pharmaceutical compositions comprising these antibodies, nucleic acids encoding these antibodies, expression vectors and host cells for preparing these antibodies, and methods for treating subjects using these antibodies are also provided. The antibodies disclosed herein are particularly useful for enhancing immune responses against tumor antigens and therefore for treating cancer in subjects. All instances of "isolated antibodies" described herein are further contemplated as antibodies that can, but do not necessarily, be isolated. All instances of "isolated polynucleotides" described herein are further contemplated as polynucleotides that can, but do not necessarily, be isolated. All instances of "antibodies" described herein are further contemplated as antibodies that can, but do not necessarily, be isolated. All instances of "polynucleotides" described herein are further contemplated as polynucleotides that can, but do not necessarily, be isolated.

[0617] 5.1 Definition

[0618] As used herein, the terms “about” and “approximately” when used to modify numerical values ​​or ranges indicate deviations of 5% to 10% (e.g., up to 5% to 10%) above and 5% to 10% (e.g., down to 5% to 10%) from the stated value or range, which remain within the intended meaning of the stated value or range.

[0619] As used herein, the term "CD73" (also known as extracellular 5'-nucleotidase, extracellular 5'-NT, 5'-NT, and NT5E) refers to a protein encoded by the NT5E gene in humans. As used herein, the term "human CD73" refers to the CD73 protein or the extracellular domain of such proteins encoded by the human NT5E gene (e.g., the wild-type human NT5E gene). Exemplary wild-type human CD73 proteins are provided by GenBank. TM Accession numbers NP_002517.1, NP_001191742.1, AAH65937, and EAW48635 are provided. For this definition and other protein definitions herein, those skilled in the art will understand that a gene-encoded protein may be in its mature form, for example, with its signal peptide removed. Exemplary amino acid sequences of the immature human CD73 protein are provided as SEQ ID NO:99, 100, 129, and 130. Exemplary amino acid sequences of the mature human CD73 protein are amino acid residues Trp27-Gln574 of SEQ ID NO:99 and amino acid residues Trp27-Gln524 of SEQ ID NO:100. Exemplary amino acid sequences of the extracellular domains of the mature human CD73 protein are amino acid residues Trp27-Lys547 of SEQ ID NO:99 and Trp27-Lys497 of SEQ ID NO:100.

[0620] As used herein, the terms “transforming growth factor β” and “TGFβ” refer to any TGFβ family protein encoded by the TGFB1, TGFB2, and TGFB3 genes in humans. As used herein, the term “human TGFβ1” refers to the TGFβ1 protein encoded by the human TGFB1 gene (e.g., the wild-type human TGFB1 gene). Exemplary wild-type human TGFβ1 proteins are provided by GenBank. TM Accession number NP_000651.3 is provided. As used herein, the term "human TGFβ2" refers to the TGFβ2 protein encoded by the human TGFB2 gene (e.g., the wild-type human TGFB2 gene). Exemplary wild-type human TGFβ2 proteins are provided by GenBank. TM Accessions NP_001129071.1 and NP_003229.1 are provided. As used herein, the term "human TGFβ3" refers to the TGFβ3 protein encoded by the human TGFB3 gene (e.g., the wild-type human TGFB3 gene). Exemplary wild-type human TGFβ3 proteins are provided by GenBank. TM Login numbers NP_003230.1, NP_001316868.1 and NP_001316867.1 are provided.

[0621] As used herein, the terms “transforming growth factor β receptor” and “TGFβ receptor” refer to any TGFβ receptor family protein encoded by the TGFBR1, TGFBR2, and TGFBR3 genes in humans. As used herein, the term “human TGFβR1” refers to the TGFβR1 protein encoded by the human TGFBR1 gene (e.g., the wild-type human TGFBR1 gene). Exemplary wild-type human TGFβR1 proteins are available from GenBank. TM Accessions NP_004603.1, NP_001124388.1, and NP_001293139.1 are provided. As used herein, the term "human TGFβ2" refers to the TGFβ2 protein encoded by the human TGFB2 gene (e.g., the wild-type human TGFB2 gene). Exemplary wild-type human TGFβ2 proteins are provided by GenBank. TM Accessions NP_001129071.1 and NP_003229.1 are provided. As used herein, the term "human TGFβ3" refers to the TGFβ3 protein encoded by the human TGFB3 gene (e.g., the wild-type human TGFB3 gene). Exemplary wild-type human TGFβ3 proteins are provided by GenBank. TM Login numbers NP_003230.1, NP_001316868.1 and NP_001316867.1 are provided.

[0622] As used herein, the terms “vascular endothelial growth factor” and “VEGF” refer to any VEGF family protein in humans encoded by the VEGFA, VEGFB, VEGFC, and VEGFD genes. As used herein, the term “human VEGF-A” refers to the VEGF-A protein encoded by the human VEGFA gene (e.g., the wild-type human VEGFA gene). Exemplary wild-type human VEGF-A proteins are provided by GenBank. TM Registration numbers NP_001020537.2, NP_001020538.2, NP_001020539.2, NP_001020540.2, NP_001020541 .2, NP_001028928.1, NP_001165093.1, NP_001165094.1, NP_001165095.1, NP_001165096. 1. NP_001165097.1, NP_001165098.1, NP_001165099.1, NP_001165100.1, NP_001165101.1, NP_001191313.1, NP_001191314.1, NP_001273973.1, NP_001303939.1, and NP_003367.4 are provided. As used herein, the term "human VEGF-B" refers to the VEGF-B protein encoded by the human VEGFB gene (e.g., the wild-type human VEGFB gene). Exemplary wild-type human VEGF-B proteins are provided by GenBank. TM Accessions NP_001230662.1 and NP_003368.1 are provided. As used herein, the term "human VEGF-C" refers to the VEGF-C protein encoded by a human VEGFC gene (e.g., the wild-type human VEGFC gene). Exemplary wild-type human VEGF-C proteins are provided by GenBank. TM Accession number NP_005420.1 is provided. As used herein, the term "human VEGF-D" refers to the VEGF-D protein encoded by the human VEGFD gene (e.g., the wild-type human VEGFD gene). Exemplary wild-type human VEGF-D proteins are provided by GenBank. TM Login ID NP_004460.1 is provided.

[0623] As used herein, the terms “vascular endothelial growth factor receptor” and “VEGFR” refer to any VEGF receptor family protein encoded in humans by the FLT1 (also known as VEGFR1), KDR (also known as VEGFR2), and FLT4 (also known as VEGFR3) genes. As used herein, the term “human VEGFR1” refers to the VEGFR1 protein encoded by the human FLT1 gene (e.g., the wild-type FLT1 gene). Exemplary wild-type human VEGFR1 proteins are available from GenBank. TM Accessions NP_002010.2, NP_001153392.1, NP_001153502.1, and NP_001153503.1 are provided. As used herein, the term “human VEGFR2” refers to the VEGFR2 protein encoded by the human KDR gene (e.g., the wild-type human KDR gene). Exemplary wild-type human VEGFR2 proteins are provided by GenBank. TM Accession number NP_002244.1 is provided. As used herein, the term "human VEGFR3" refers to the VEGFR3 protein encoded by the human FLT4 gene (e.g., the wild-type human FLT4 gene). Exemplary wild-type human VEGFR3 proteins are provided by GenBank. TM Login numbers NP_891555.2, NP_002011.2 and NP_001341918.1 are provided.

[0624] As used herein, the term "antibody" includes full-length antibodies, antigen-binding fragments of full-length antibodies, and molecules containing antibody CDR, VH region, and / or VL region. Examples of antibodies include, but are not limited to, monoclonal antibodies, antibodies produced in a recombinant manner, monospecific antibodies, multispecific antibodies (including bispecific antibodies), human antibodies, humanized antibodies, chimeric antibodies, immunoglobulins, synthetic antibodies, tetrameric antibodies containing two heavy chain and two light chain molecules, antibody light chain monomers, antibody heavy chain monomers, antibody light chain dimers, antibody heavy chain dimers, antibody light chain-antibody heavy chain pairs, endoantibodies, heteroconjugated antibodies, antibody-drug conjugates, single-domain antibodies, monovalent antibodies, single-chain antibodies or single-chain Fv (scFv), camelified antibodies, affinity molecules, Fab fragments, F(ab')2 fragments, disulfide-linked Fv (sdFv), anti-idiotypic (anti-Id) antibodies (including, for example, anti-anti-Id antibodies), and antigen-binding fragments of any of the above, as well as conjugates or fusion proteins containing any of the above. In some embodiments, the antibodies described herein refer to a population of polyclonal antibodies. Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, or IgY), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, or IgA2), or any subclass (e.g., IgG). 2a or IgG 2bThe antibody is an immunoglobulin molecule. In some embodiments, the antibody described herein is an IgG antibody or a class (e.g., human IgG1 or IgG4) or subclass. In one specific embodiment, the antibody is a humanized monoclonal antibody. In another specific embodiment, the antibody is a human monoclonal antibody. In some embodiments, the antibody as described herein comprises a full-length antibody (e.g., a full-length antibody that specifically binds to CD73) or an antigen-binding fragment thereof, which is linked to a ligand-binding moiety that specifically binds to molecules other than CD73 (e.g., a ligand-binding moiety that specifically binds to TGFβ or VEGF, such as a ligand-binding moiety containing one or more extracellular domains of a TGFβ receptor or VEGF receptor).

[0625] As used herein, the terms “VH region” and “VL region” refer to the variable regions of a single antibody heavy and light chains containing FR (framework regions) 1, 2, 3 and 4 and CDR (complementarity-determining regions) 1, 2 and 3, respectively (see Kabat et al., (1991) Sequences of Proteins of Immunological Interest (NIH Publication No. 91-3242, Bethesda), which is incorporated herein by reference in its entirety).

[0626] As used herein, the term “CDR” or “complementarity-determining region” refers to a discontinuous antigen-binding site found within the variable region of both the heavy and light chain polypeptides. These specific regions have been described by the following: Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991), Chothia et al., J. Mol. Biol. 196: 901-917 (1987) and MacCallum et al., J. Mol. Biol. 262: 732-745 (1996), all of which are incorporated herein by reference in their entirety, wherein, when compared with each other, the definition includes overlap or subgroups of amino acid residues. In some embodiments, the term "CDR" is as defined by MacCallum et al., J. Mol. Biol. 262:732-745 (1996) and Martin A. "Protein Sequence and Structure Analysis of Antibody Variable Domains," in Antibody Engineering, Kontermann and Dübel, eds., Chapter 31, pp.422-439, Springer-Verlag, Berlin (2001). In some embodiments, the term "CDR" is as defined by Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991). In some implementations, heavy chain CDRs and / or light chain CDRs are defined by performing structural analysis on the antibody and identifying residues in the variable region that are predicted to contact the epitope region of the target molecule (e.g., human and / or cynomolgus monkey CD73). CDRH1, CDRH2, and CDRH3 represent heavy chain CDRs, while CDRL1, CDRL2, and CDRL3 represent light chain CDRs.

[0627] As used herein, the term “framework (FR) amino acid residue” refers to those amino acids in the frame region of the antibody variable region. As used herein, the terms “frame region” or “FR region” include amino acid residues that are part of the variable region but not part of the CDR (e.g., as defined by Kabat or MacCallum using the CDR).

[0628] As used herein, the terms “variable region” and “variable domain” are used interchangeably and are common in the art. A variable region typically refers to a portion of an antibody, generally a portion of the light or heavy chain, typically about 110 to 120 or 110 to 125 amino acids from the amino terminus of the mature heavy chain and about 90 to 115 amino acids from the mature light chain, which varies considerably in sequence between antibodies and is used in the binding and specificity of a particular antibody to its specific antigen. Sequence variability is concentrated in those regions called complementarity-determining regions (CDRs), while more highly conserved regions within a variable domain are called frame regions (FRs). Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for antibody-antigen interactions and specificity. In some embodiments, the variable region is a human variable region. In some embodiments, the variable region comprises a rodent or mouse CDR and a human frame region (FR). In certain embodiments, the variable region is a primate (e.g., a non-human primate) variable region. In some implementations, the variable region includes the rodent or mouse CDR and the primate (e.g., non-human primate) frame region (FR).

[0629] The terms “VL” and “VL domain” are used interchangeably to refer to the variable region of the light chain of an antibody.

[0630] The terms “VH” and “VH domain” are used interchangeably to refer to the variable region of the heavy chain of an antibody.

[0631] As used herein, the terms “constant region” and “constant domain” are interchangeable and common in the art. A constant region is an antibody portion that does not directly participate in antibody-antigen binding but can exhibit various effector functions, such as interaction with Fc receptors (e.g., Fcγ receptors), or the carboxyl-terminal portion of the light and / or heavy chains. The constant regions of immunoglobulin molecules typically have a more conserved amino acid sequence than the variable domains of immunoglobulins.

[0632] As used herein, the term “heavy chain” when used in relation to antibodies can refer to any different type of amino acid sequence based on a constant structural domain, such as alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), which respectively produce the IgA, IgD, IgE, IgG, and IgM classes of antibodies, including subclasses of IgG such as IgG1, IgG2, IgG3, and IgG4.

[0633] As used herein, the term "light chain" when used in relation to an antibody can refer to any unique type of amino acid sequence based on a constant domain, such as kappa(κ) or lambda(λ). Light chain amino acid sequences are well known in the art. In a particular embodiment, the light chain is a human light chain.

[0634] As used herein, the term “EU numbering system” refers to the EU numbering convention for constant regions of antibodies as described in the following: Edelman, G.M. et al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and Kabat et al., Sequences of Proteins of Immunological Interest, USDept. Health and Human Services, 5th edition, 1991, each of which is incorporated herein by reference in its entirety.

[0635] As used herein, the term "binding moiety" refers to a portion having the ability to specifically bind to a target molecule or complex. Binding moieties may comprise small molecules, peptides, modified peptides (e.g., peptides having non-natural amino acid residues and / or stapled peptides), polypeptides, proteins, antibodies, antibody fragments, scFvs, Fc-containing polypeptides, fusion antibodies, ligands, aptamers, nucleic acids, variants thereof, or any combination thereof. In some embodiments, the binding moiety comprises a domain of a native protein or a variant thereof, via which the native protein binds to the target molecule or complex. For example, the term "TGFβ binding moiety" refers to a portion that specifically binds to one or more proteins of the TGFβ family (e.g., TGFβ1, TGFβ2, or TGFβ3), and in some embodiments may comprise an extracellular domain of a TGFβ receptor (e.g., TGFβR1, TGFβR2, or TGFβR3). The term "VEGF binding moiety" refers to a portion that specifically binds to one or more proteins of the VEGF family (e.g., VEGF-A, VEGF-B, VEGF-C, or VEGF-D), and in some embodiments may include an extracellular domain of a VEGF receptor (e.g., VEGFR1, VEGFR2, or VEGFR3).

[0636] "Binding affinity" generally refers to the total strength of the non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen). Unless otherwise indicated, "binding affinity" as used herein refers to the endogenous binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., antibody and antigen). The affinity of molecule X for its partner Y can generally be expressed using the dissociation constant (K). D Affinity can be measured and / or represented in a variety of ways known in the art, including but not limited to the equilibrium dissociation constant (K). D ) and equilibrium association constant (K A K D It is k off / k on The quotient is calculated, and KA It is k on / k off The quotient is calculated. k on This refers to, for example, the association rate constant between an antibody and an antigen, and k off This refers to, for example, the dissociation rate constant between an antibody and an antigen. k on and k off This can be achieved using techniques known to those skilled in the art, such as... Or determine using KinExA. As used in this article, "lower affinity" refers to a larger K... D .

[0637] As used herein, the term “maximum binding” refers to the maximum signal (expressed in resonance units (RU) observed in the binding of an antigen (e.g., CD73 protein) to an antibody (e.g., anti-CD73 antibody) at a given concentration in a surface plasmon resonance binding assay (e.g., the surface plasmon resonance binding assay described in Section 6.5.2 of this document).

[0638] As used herein in the context of binding affinity, if, for example, in a given experiment or using the average of multiple experiments (as assessed by, for example, a binding affinity of the test antibody to the first antigen is reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% relative to the binding affinity of the test antibody to the second antigen, then the binding affinity of the test antibody to the first antigen is “significantly reduced” relative to the binding affinity of the test antibody to the second antigen. As used herein in the context of maximum binding, if, for example, in a given experiment or using the average of multiple experiments (as assessed by, for example, a binding affinity assay disclosed herein), the maximum binding between the test antibody and the first antigen is reduced by at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% relative to the maximum binding between the test antibody and the second antigen, then the maximum binding between the test antibody and the first antigen is “significantly reduced” relative to the maximum binding between the test antibody and the second antigen.

[0639] As used herein, the terms “specific binding,” “specific recognition,” “immunospecific binding,” and “immunospecific recognition” are similar terms in the context of antibodies and refer to molecules that bind to antigens (e.g., epitopes or immune complexes) in a manner that is strictly speaking understood by one of ordinary skill in the art. For example, molecules that specifically bind to antigens may bind to other peptides or polypeptides, typically with lower affinity, such as through methods like immunoassays. The assay is performed using a KinExA 3000 instrument (Sapidyne Instruments, Boise, ID) or other assays known in the art. In one specific embodiment, a molecule that specifically binds to an antigen is measured at K when the molecule nonspecifically binds to another antigen. A At least 2 log (i.e., a coefficient of 10), 2.5 log, 3 log, 4 log or greater K A Binding to antigens. Those skilled in the art will understand that antibodies as described herein can specifically bind to more than one antigen (e.g., via different regions of the antibody molecule). For example, antibodies as described herein can specifically bind to CD73 and TGFβ or specifically bind to CD73 and VEGF.

[0640] As used herein, the term "immunoglobulin chain" refers to a polypeptide or antigen-binding fragment of an antibody containing antibody VH or VL. An immunoglobulin chain may contain, for example, VH, VL, a full-length antibody heavy chain, a full-length antibody light chain, or an antigen-binding fragment of any of the foregoing.

[0641] As used herein, the term “linked to” refers to a covalent or non-covalent bond between two molecules or portions. Those skilled in the art will understand that when a first molecule or portion is linked to a second molecule or portion, the link need not be direct, but can be via an intermediate molecule or portion. For example, when the heavy chain variable region of a full-length antibody is linked to a ligand-binding moiety, the ligand-binding moiety can bind to the constant region of the full-length antibody (e.g., the heavy chain constant region) (e.g., via a peptide bond), rather than directly to the heavy chain variable region.

[0642] In another specific embodiment, the molecule specifically binding to the antigen does not cross-react with other proteins under similar binding conditions. In another specific embodiment, the molecule specifically binding to CD73 does not cross-react with other non-CD73 proteins. In one specific embodiment, this document provides an antibody that binds to CD73 (e.g., human CD73) with a higher affinity than that against another unrelated antigen. In some embodiments, this document provides an antibody that, as measured by, for example, radioimmunoassay, surface plasmon resonance, or dynamic exclusion assay, binds to CD73 (e.g., human CD73) with an affinity 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or higher than that binding to another unrelated antigen. In one specific implementation, the anti-CD73 antibody described herein binds to less than 10%, 15%, or 20% of the antibody binding to the CD73 protein, as measured by, for example, radioimmunoassay.

[0643] As used herein, "epitope" is a term in the art and refers to a localized antigenic region that can specifically bind to an antibody. An epitope can be, for example, a continuous amino acid of a polypeptide (linear or continuous epitope), or an epitope can be, for example, an aggregation of two or more discontinuous regions of one or more polypeptides (conformal, nonlinear, discontinuous, or noncontinuous epitopes). In some embodiments, the epitope to which the antibody binds can be determined by, for example, NMR spectroscopy, X-ray diffraction crystallography, ELISA assays, hydrogen / deuterium exchange combined mass spectrometry (e.g., liquid chromatography-electrospray ionization mass spectrometry), array-based oligopeptide scanning assays (e.g., using CLIPS (peptide chemically linked to the backbone) to locate discontinuous or conformational epitopes of restricted peptides), and / or mutagenic localization (e.g., site-directed mutagenic localization). In X-ray crystallography, crystallization can be achieved using any of the methods known in the art (e.g., Giegé R et al., (1994) Acta Crystallogr D Biol Crystallogr 50 (Pt4):339-350; McPherson A (1990) Eur J Biochem 189:1-23; Chayen NE (1997) Structure 5:1269-1274; McPherson A (1976) J Biol Chem 251:6300-6303, each of which is incorporated herein by reference in its entirety). Antibody: Antigen crystals can be studied using well-known X-ray diffraction techniques and refined using computer software such as X-PLOR (Yale University, 1992, published by Molecular Simulations, Inc.; see, for example, Meth Enzymol (1985) volumes 114 & 115, eds. Wyckoff HW et al.; US2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49 (Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed. Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56 (Pt 10): 1316-1323), each of which is incorporated herein by reference in its entirety. Mutagenesis localization studies can be performed using any method known to those skilled in the art.For descriptions of mutagenesis techniques (including alanine scanning mutagenesis), see, for example, Champe M et al., (1995) J Biol Chem 270:1388-1394 and Cunningham BC & Wells JA (1989) Science 244:1081-1085, each of which is incorporated herein by reference in its entirety. CLIPS (Chemical Linking of Peptides to Scaffolds) is a technique for presenting one or more peptides in a structurally restricted configuration to act as functional mimics of complex protein domains. See, for example, U.S. Publications US2008 / 0139407 A1 and US2007 / 099240 A1 and U.S. Patent No. 7,972,993, each of which is incorporated herein by reference in its entirety. In one specific embodiment, alanine scanning mutagenesis studies are used to determine the epitopes of an antibody. In one specific embodiment, the epitopes of an antibody are determined using hydrogen / deuterium exchange combined mass spectrometry. In one specific implementation, the antibody epitope is determined using CLIPS epitope localization technology from Pepscan Therapeutics. In another specific implementation, the antibody epitope is determined by protein mutagenesis, for example by generating a transgenic mutant of the antigen with a portion of an ortholog of the antigen from another species and then testing for the loss of binding of the transgenic mutant to the antibody (e.g., by a cell binding assay based on FACS as described herein).

[0644] As used herein, the term "epitope located within a region of human CD73" refers to an epitope comprising one or more residues of the amino acid residues within the designated region. In some embodiments, the epitope comprises every single amino acid residue within the designated region. In some embodiments, the epitope consists of every single amino acid residue within the designated region. In some embodiments, one or more additional amino acid residues of human CD73 outside the designated region bind to the antibody along with the epitope located within the designated region.

[0645] As used herein, the terms “T cell receptor” and “TCR” are used interchangeably and refer to full-length heterodimeric αβ or γδ TCRs, antigen-binding fragments of full-length TCRs, and molecules containing TCR CDRs or variable regions. Examples of TCRs include, but are not limited to, full-length TCRs, antigen-binding fragments of full-length TCRs, soluble TCRs without transmembrane and cytoplasmic regions, single-chain TCRs containing variable regions of TCRs attached by flexible linkers, TCR chains linked by engineered disulfide bonds, monospecific TCRs, multispecific TCRs (including bispecific TCRs), TCR fusions, human TCRs, humanized TCRs, chimeric TCRs, recombinantly generated TCRs, and synthetic TCRs. The terms encompass wild-type TCRs and genetically engineered TCRs (e.g., chimeric TCRs containing chimeric TCR chains, which include a first portion of a TCR from a first species and a second portion of a TCR from a second species).

[0646] As used herein, the terms “major histocompatibility complex” and “MHC” are used interchangeably and refer to MHC class I molecules and / or MHC class II molecules.

[0647] As used herein, the term “peptide-MHC complex” refers to an MHC molecule (MHC class I or MHC class II) in a peptide-binding pocket recognized in the art, in which a peptide is bound to an MHC.

[0648] As used herein, the term “treatment” refers to the therapeutic or preventative measures described herein. A “treatment” method involves administering antibodies to a subject who has a disease or condition or is susceptible to such a disease or condition in order to prevent, cure, delay, reduce the severity of, or improve one or more symptoms of said disease or condition or recurrent disease or condition, or to prolong the subject’s survival beyond the expected survival without such treatment.

[0649] As used herein, the term "effective amount" in the context of administering a therapy to a subject refers to the amount of therapy that achieves the desired preventive or therapeutic effect.

[0650] As used herein, the term "subject" includes any human or non-human animal. In one embodiment, the subject is a human or non-human mammal. In another embodiment, the subject is a human.

[0651] The determination of the "percentage of similarity" between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be achieved using mathematical algorithms. A specific, non-limiting example of a mathematical algorithm for comparing two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87:2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90:5873-5877, each of which is incorporated herein by reference in its entirety. Such algorithms are incorporated in the NBLAST and XBLAST procedures of Altschul SF et al., (1990) J Mol Biol 215:403 (which is incorporated herein by reference in its entirety). BLAST nucleotide searches can be performed using the NBLAST nucleotide procedure parameter set, e.g., score = 100, word length = 12, to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed using the XBLAST procedure parameter set, for example, a score of 50 and a word length of 3, to obtain amino acid sequences homologous to the protein molecules described herein. For comparison purposes, vacancy-based alignments can be used, as described in Altschul SF et al., (1997) Nuc Acids Res 25:3389-3402 (which is incorporated herein by reference in its entirety). Alternatively, PSI BLAST can be used for iterative searches that detect long-distance relationships between molecules (ibid.). When using BLAST, vacancy-based BLAST, and PSI BLAST procedures, the default parameters of the respective procedures (e.g., XBLAST and NBLAST) can be used (see, for example, the World Wide Web of the National Center for Biotechnology Information (NCBI), ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm for comparing sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17 (which is incorporated herein by reference in its entirety). Such algorithms are incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When comparing amino acid sequences using the ALIGN program, the PAM120 weighted residue table, vacancy length penalty 12, and vacancy penalty 4 can be used.

[0652] The percentage of consistency between two sequences can be determined using techniques similar to those described above, with or without gaps allowed. When calculating the percentage of consistency, typically only exact matches are counted.

[0653] As used herein, the term "lipid nanoparticle" refers to one or more spherical nanoparticles having an average diameter between about 10 and about 1000 nanometers and comprising a solid lipid core matrix capable of dissolving lipophilic molecules. In some embodiments, the lipid core is stabilized by a surfactant (e.g., an emulsifier) ​​and may comprise one or more, including combinations thereof, of triglycerides (e.g., glyceryl tristearate), diglycerides (e.g., glycerol bahenate), monoglycerides (e.g., glyceryl monostearate), fatty acids (e.g., stearic acid), steroids (e.g., cholesterol), and waxes (e.g., hexadecyl palmitate). Lipid nanoparticles are described, for example, in Petrilli et al., Curr PharmBiotechnol. 15:847-55, 2014; and in U.S. Patent Nos. 6,217,912; 6,881,421; 7,402,573; 7,404,969; 7,550,441; 7,727,969; 8,003,621; 8,691,750; 8,871,509; 9,017,726; 9,173,853; 9,220,779; 9,227,917; and 9,278,130, each of which is incorporated herein by reference in its entirety.

[0654] 5.2 Anti-CD73 antibody

[0655] In one aspect, this disclosure provides antibodies that specifically bind to CD73 (e.g., human CD73, cynomolgus monkey CD73, or mouse CD73) and antagonize CD73 function. The amino acid sequences of exemplary antibodies are listed in Table 1 herein.

[0656] Table 1. Amino acid sequences of exemplary anti-CD73 antibodies

[0657]

[0658]

[0659]

[0660]

[0661]

[0662]

[0663]

[0664]

[0665]

[0666]

[0667]

[0668]

[0669]

[0670]

[0671]

[0672]

[0673] Table 2. Exemplary heavy chain CDR sequences of anti-CD73 antibodies

[0674]

[0675]

[0676] V in Table 2 H CDR is determined based on Kabat.

[0677] Table 3. Exemplary light chain CDR sequences of anti-CD73 antibodies

[0678]

[0679]

[0680] V in Table 3 L CDR is determined based on Kabat.

[0681] Table 4. Exemplary anti-CD73 antibody VH, VL, full-length heavy chain, and full-length light chain sequences*

[0682]

[0683]

[0684] *In this table, if the polypeptide of an anti-CD73 antibody containing a ligand-binding moiety contains a full-length heavy chain, it is called a full-length heavy chain.

[0685] Table 5. Exemplary sequences of CD73 and its family members

[0686]

[0687]

[0688]

[0689]

[0690]

[0691]

[0692]

[0693]

[0694] Table 6. Exemplary sequences of linkers, ligand-binding moieties, and anti-CD73 antibodies containing ligand-binding moieties*

[0695]

[0696]

[0697]

[0698]

[0699]

[0700]

[0701] *In this table, if the polypeptide of an anti-CD73 antibody containing a ligand-binding moiety contains a full-length heavy chain, it is called a full-length heavy chain.

[0702] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), the antibodies comprising a VH domain including one, two, or all three CDRs of the VH domains listed in Table 1 herein. In some embodiments, the antibody comprises CDRH1 of the VH domains listed in Table 1. In some embodiments, the antibody comprises CDRH2 of the VH domains listed in Table 1. In some embodiments, the antibody comprises CDRH3 of the VH domains listed in Table 1.

[0703] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), the antibodies comprising a VL domain including one, two, or all three CDRs of the VL domains listed in Table 1 herein. In some embodiments, the antibody comprises CDRL1 of the VL domains listed in Table 1. In some embodiments, the antibody comprises CDRL2 of the VL domains listed in Table 1. In some embodiments, the antibody comprises CDRL3 of the VL domains listed in Table 1.

[0704] In some embodiments, the CDR of the antibody can be determined according to Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest (1991), each of which is incorporated herein by reference in its entirety. In some embodiments, the light chain CDR of the antibody is determined according to Kabat and the heavy chain CDR of the antibody is determined according to MacCallum (ibid.). In some embodiments, the heavy chain CDR and / or light chain CDR are defined by structural analysis of the antibody and identification of residues in the variable region predicted to contact the epitope region of the target molecule (e.g., human and / or cynomolgus monkey CD73).

[0705] In some implementations, the CDR of the antibody can be determined according to the Chothia numbering scheme, which refers to the location of the immunoglobulin structural loop (see, for example, Chothia C & Lesk AM, (1987), J Mol Biol 196:901-917; Al-Lazikani B et al., (1997) J Mol Biol 273:927-948; Chothia C et al., (1992) J Mol Biol 227:799-817; Tramontano A et al., (1990) J Mol Biol 215(1):175-82; and U.S. Patent No. 7,709,226, all of which are incorporated herein by reference in their entirety). Typically, when using the Kabat numbering convention, the Chothia CDRH1 ring is located at positions 26 to 32, 33, or 34 of the heavy chain amino acids; the Chothia CDRH2 ring is located at positions 52 to 56 of the heavy chain amino acids; and the Chothia CDRH3 ring is located at positions 95 to 102 of the heavy chain amino acids. Conversely, the Chothia CDRL1 ring is located at positions 24 to 34 of the light chain amino acids; the Chothia CDRL2 ring is located at positions 50 to 56 of the light chain amino acids; and the Chothia CDRL3 ring is located at positions 89 to 97 of the light chain amino acids. When using the Kabat numbering convention, the end of the Chothia CDRH1 ring varies between H32 and H34 depending on the ring length (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B is present, the ring terminates at 32; if only 35A is present, the ring terminates at 33; and if both 35A and 35B are present, the ring terminates at 34).

[0706] In some implementations, the CDR of the antibody can be determined based on the determination of the antibody's CDR. MacCallum RM et al., (1996) J MolBiol 262:732-745, which is incorporated herein by reference in its entirety. See also, for example, Martin A. “Protein Sequence and Structure Analysis of Antibody Variable Domains,” in Antibody Engineering, Kontermann and Dübel, eds., Chapter 31, pp.422-439, Springer-Verlag, Berlin (2001), which is incorporated herein by reference in its entirety.

[0707] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), comprising the Chothia VH CDR of VH disclosed in Table 1 herein. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), comprising the Chothia VL CDR of VL disclosed in Table 1 herein. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), comprising the Chothia VH CDR and Chothia VL CDR of the antibodies disclosed in Table 1 herein. In some embodiments, the antibody that specifically binds to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) comprises one or more CDRs, wherein the Chothia and Kabat CDRs have the same amino acid sequence. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) and comprise a combination of Kabat CDR and Chothia CDR.

[0708] In some implementations, the CDR of the antibody can be determined according to the IMGT numbering system as described in Lefranc MP, (1999) The Immunologist 7:132-136 and Lefranc MP et al., (1999) Nucleic Acids Res 27:209-212, each of which is incorporated herein by reference in its entirety. According to the IMGT numbering scheme, CDRH1 is at positions 26 to 35, CDRH2 at positions 51 to 57, CDRH3 at positions 93 to 102, CDRL1 at positions 27 to 32, CDRL2 at positions 50 to 52, and CDRL3 at positions 89 to 97. IMGT unique codes can also be defined as in Lefranc MP et al., (2009) Nucleic Acids Res 37:D1006-D1012 and in the IMGT database.

[0709] In some embodiments, this disclosure provides antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) and contain the CDR of the antibodies disclosed herein as determined by the IMGT numbering system, for example, as described in Lefranc MP (1999) ibid. and Lefranc MP et al., (1999) ibid. or Lefranc MP et al., (2009) ibid.

[0710] In some embodiments, the CDR of the antibody can be determined according to the AbM numbering scheme, which refers to the AbM hypervariable region, representing a trade-off between the Kabat CDR and the Chothia structural loop, and is used by the AbM antibody modeling software of Oxford Molecular (Oxford Molecular Group, Inc.) (which is incorporated herein by reference in its entirety). In one specific embodiment, this disclosure provides an antibody that specifically binds to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) and comprises the CDR of the antibody disclosed herein as determined by the AbM numbering scheme.

[0711] In some embodiments, the CDR of the antibody can be determined according to the AHo numbering system (as described in Honegger and Plückthun, A., J. Mol. Biol. 309:657-670 (2001), which is incorporated herein by reference in its entirety). In some embodiments, this disclosure provides antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) and comprise the CDR of the antibodies disclosed herein as determined by the AHo numbering scheme.

[0712] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising the amino acid sequences of the CDRH1, CDRH2, and CDRH3 regions of the VH domain shown in SEQ ID NO: 19, 20, 21, 23, 24, 25, 26, 27, 28, 29, or 30, and the light chain variable region comprising the amino acid sequences of the CDRL1, CDRL2, and CDRL3 regions of the VL domain shown in SEQ ID NO: 33, 34, 35, 37, 38, 39, or 40, wherein each CDR is defined according to the MacCallum definition, Kabat definition, Chothia definition, IMGT numbering system, AbM definition, structural analysis, or a combination thereof, wherein the structural analysis identifies residues in the variable region that are predicted to contact epitope regions of CD73 (e.g., human, mouse, or cynomolgus monkey CD73). In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region containing the amino acid sequences of the CDRH1, CDRH2, and CDRH3 regions comprising a VH domain and a light chain variable region containing the amino acid sequences of the CDRL1, CDRL2, and CDRL3 regions comprising a VL domain, wherein the VH domain and VL domain each comprise SEQ ID [SEQ ID]. NO:19 and 33; 20 and 34; 21 and 35; 23 and 37; 24 and 37; 25 and 37; 26 and 37; 27 and 37; 28 and 37; 29 and 37; 30 and 37; 23 and 38; 24 and 38; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 23 and 39; or 23 and 40, wherein each CDR is defined according to the MacCallum definition, Kabat definition, Chothia definition, IMGT numbering system, AbM definition, structural analysis, or a combination thereof, wherein the structural analysis identifies residues in the variable region that are predicted to contact the epitope region of CD73 (e.g., human, mouse, or cynomolgus monkey CD73). In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) and comprise a combination of a CDR defined by Kabat and a CDR defined by structural analysis of the antibody, wherein the structural analysis identifies residues in the variable region that are predicted to contact the epitope region of CD73 (e.g., human, mouse, or cynomolgus monkey CD73).

[0713] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), comprising one or more of the six CDRs disclosed herein as defined by antibody numbering schemes such as Kabat, IMGT (e.g., IMGT Unique), AHo, or Chothia. In some embodiments, the antibody comprises at least one, at least two, at least three, at least four, or at least five CDRs present in the BA025 antibody. The Kabat, IMGT Unique, Aho, and Chothia CDRs for the BA025 antibody are provided in Table 1 herein.

[0714] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), comprising one or more of the six CDRs disclosed herein as defined by antibody numbering schemes such as Kabat, IMGT (e.g., IMGT Unique), AHo, or Chothia. In some embodiments, the antibody comprises at least one, at least two, at least three, at least four, or at least five CDRs present in the BA020 antibody. The Kabat, IMGT Unique, Aho, and Chothia CDRs for the BA020 antibody are provided in Table 1 herein.

[0715] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), the antibodies comprising:

[0716] (a) CDRH1 contains the amino acid sequence X1X2WX3X4 (SEQ ID NO:13), in which

[0717] X1 is S or N;

[0718] X2 is either S or Y;

[0719] X3 is either I or M; and

[0720] X4 is N or H;

[0721] (b) CDRH2 contains the amino acid sequence X1IYPRX2X3DTNYX4X5KFKX6 (SEQ ID NO:14), in which

[0722] X1 is either R or T;

[0723] X2 is N, A, or S;

[0724] X3 is either G or S;

[0725] X4 is N, A, or S;

[0726] X5 is either G or Q; and

[0727] X6 is either D or G;

[0728] (c) CDRH3 contains the amino acid sequence of LLDYSMDY (SEQ ID NO:7);

[0729] (d) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0730] X1 is N or I; and

[0731] X2 is either Y or S;

[0732] (e) CDRL2 contains the amino acid sequence YTSRLHS (SEQ ID NO: 10); and / or

[0733] (f) CDRL3 contains the amino acid sequence QQGNTLPXT (SEQ ID NO:17), in which

[0734] X is either L or W.

[0735] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), the antibodies comprising:

[0736] (a) CDRH1 contains the amino acid sequence of SSWIN (SEQ ID NO:2);

[0737] (b) CDRH2 contains the amino acid sequence RIYPRX1GDTNYX2GKFKD (SEQ ID NO:15), in which

[0738] X1 is N, A, or S; and

[0739] X2 is N, A, or S;

[0740] (c) CDRH3 contains the amino acid sequence of LLDYSMDY (SEQ ID NO:7);

[0741] (d) CDRL1 contains the amino acid sequence RASQDISX1X2LN (SEQ ID NO:16), in which

[0742] X1 is N or I; and

[0743] X2 is either Y or S;

[0744] (e) CDRL2 contains the amino acid sequence YTSRLHS (SEQ ID NO: 10); and / or

[0745] (f) CDRL3 contains the amino acid sequence QQGNTLPLT (SEQ ID NO:12).

[0746] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VH domains comprising the amino acid sequences of CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO:1, 3, and 7, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VH domains comprising the amino acid sequences of CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO:2, 4, and 7, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VH domains comprising the amino acid sequences of CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO:2, 5, and 7, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VH domains comprising the amino acid sequences of CDRH1, CDRH2, and CDRH3 shown in SEQ ID NO:2, 6, and 7, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VL domains comprising the amino acid sequences of CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO:8, 10, and 11, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VL domains comprising the amino acid sequences of CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO:9, 10, and 12, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise VL domains comprising the amino acid sequences of CDRL1, CDRL2, and CDRL3 shown in SEQ ID NO:8, 10, and 12, respectively.

[0747] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO: 1, 3, 7, 8, 10, and 11, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO: 2, 4, 7, 8, 10, and 11, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO:2, 4, 7, 8, 10, and 12, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO:2, 4, 7, 9, 10, and 12, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO:2, 5, 7, 8, 10, and 12, respectively.In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO:2, 6, 7, 8, 10, and 12, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibody comprises a heavy chain variable region comprising CDRH1, CDRH2, and CDRH3 regions and a light chain variable region comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO:2, 5, 7, 9, 10, and 12, respectively. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), wherein the antibodies comprise heavy chain variable regions comprising CDRH1, CDRH2, and CDRH3 regions and light chain variable regions comprising CDRL1, CDRL2, and CDRL3 regions, wherein the CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 regions comprise the amino acid sequences shown in SEQ ID NO:2, 6, 7, 9, 10, and 12, respectively.

[0748] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:31. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO:19-21 and 23-30. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibody comprising a heavy chain variable region containing an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 19-21 and 23-30.

[0749] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 41 or 42. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a light chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 33-35 and 37-40. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibody comprising a light chain variable region containing an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 33-35 and 37-40.

[0750] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO:31 and a light chain variable region comprising an amino acid sequence of SEQ ID NO:41 or 42. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO:19-21 and 23-30 and a light chain variable region comprising an amino acid sequence selected from SEQ ID NO:33-35 and 37-40. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region and a light chain variable region, said heavy chain variable region comprising at least 75%, 80%, 85%, 90%, 95%, or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%) of an amino acid sequence selected from SEQ ID NO: 19-21 and 23-30, said light chain variable region comprising ... The amino acid sequences of NO:33-35 and 37-40 are at least 75%, 80%, 85%, 90%, 95% or 100% (e.g., at least 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99%) identical.

[0751] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region and a light chain variable region comprising, respectively, the amino acid sequences shown in SEQ ID NO: 19 and 33; 20 and 34; 21 and 35; 23 and 37; 24 and 37; 25 and 37; 26 and 37; 27 and 37; 28 and 37; 29 and 37; 30 and 37; 23 and 38; 24 and 38; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 23 and 39; or 23 and 40. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:19 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:33. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:20 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:34. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:21 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:35. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:24 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:24 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38.In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:26 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:28 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:25 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:26 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:27 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:28 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:29 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37.In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:30 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:37. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:39. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:23 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:40.

[0752] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region containing an amino acid sequence derived from a mouse IgHV1S12 germline sequence. In some embodiments, the mouse IgHV1S12 germline sequence is shown in SEQ ID NO:18. One or more regions selected from frames 1, 2, 3, CDRH1, and CDRH2 (e.g., two, three, four, or five of these regions) may be derived from the mouse IgHV1S12 germline sequence. In one embodiment, frames 1, 2, 3, CDRH1, and CDRH2 are all derived from the mouse IgHV1S12 germline sequence. In some embodiments, the heavy chain variable region comprises CDRH3 containing the amino acid sequence shown in SEQ ID NO:7.

[0753] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a light chain variable region comprising an amino acid sequence derived from a mouse IgKV10-96 germline sequence. In some embodiments, the mouse IgKV10-96 germline sequence is shown in SEQ ID NO:32. One or more regions selected from frames 1, 2, 3, CDRL1, and CDRL2 (e.g., two, three, four, or five of these regions) may be derived from the mouse IgKV10-96 germline sequence. In one embodiment, frames 1, 2, 3, CDRL1, and CDRL2 are all derived from the mouse IgKV10-96 germline sequence. In some embodiments, the light chain variable region comprises CDRL3 comprising the amino acid sequence shown in SEQ ID NO:11 or 12.

[0754] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising an amino acid sequence derived from a mouse IgHV1S12 germline sequence (e.g., SEQ ID NO: 18); and a light chain variable region comprising an amino acid sequence derived from a mouse IgKV10-96 germline sequence (e.g., SEQ ID NO: 32). In some embodiments, the heavy chain variable region comprises CDRH3 comprising the amino acid sequence shown in SEQ ID NO: 7, and the light chain variable region comprises CDRL3 comprising the amino acid sequence shown in SEQ ID NO: 11 or 12.

[0755] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a heavy chain variable region comprising an amino acid sequence derived from a human IgHV5-51 germline sequence. In some embodiments, the human IgHV5-51 germline sequence is the human IgHV5-51*02 germline sequence (e.g., comprising the amino acid sequence of SEQ ID NO: 22). One or more regions selected from frames 1, 2, 3, CDRH1, and CDRH2 (e.g., two, three, four, or five of these regions) may be derived from the human IgHV5-51 germline sequence. In one embodiment, frames 1, 2, 3, CDRH1, and CDRH2 are all derived from the human IgHV5-51 germline sequence. In some embodiments, the heavy chain variable region comprises CDRH3 comprising the amino acid sequence shown in SEQ ID NO: 7.

[0756] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibodies comprising a light chain variable region comprising an amino acid sequence derived from a human IGKV1-33 germline sequence (e.g., IGKV1-33*01, e.g., comprising the amino acid sequence of SEQ ID NO:36). One or more regions selected from frames 1, 2, 3, CDRL1, and CDRL2 (e.g., two, three, four, or five of these regions) may be derived from a human germline sequence selected from IGKV1-33 (e.g., IGKV1-33*01, e.g., comprising the amino acid sequence of SEQ ID NO:36). In one embodiment, frames 1, 2, 3, CDRL1, and CDRL2 are all derived from a human IGKV1-33 germline sequence (e.g., IGKV1-33*01, e.g., comprising the amino acid sequence of SEQ ID NO:36). In some embodiments, the light chain variable region comprises CDRL3 comprising the amino acid sequence shown in SEQ ID NO:12.

[0757] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), said antibody comprising a heavy chain variable region comprising an amino acid sequence derived from a human IgHV5-51 germline sequence (e.g., the human IgHV5-51*02 germline sequence, e.g., comprising the amino acid sequence of SEQ ID NO:22); and a light chain variable region comprising an amino acid sequence derived from a human IGKV1-33 germline sequence (e.g., IGKV1-33*01, e.g., comprising the amino acid sequence of SEQ ID NO:36). In some embodiments, the heavy chain variable region comprises CDRH3 comprising the amino acid sequence shown in SEQ ID NO:7, and the light chain variable region comprises CDRL3 comprising the amino acid sequence shown in SEQ ID NO:12.

[0758] In some embodiments, this disclosure provides isolated antibodies that cross-competitively bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) with any anti-CD73 antibody disclosed herein, comprising antibodies comprising the amino acid sequences of the heavy chain variable region and light chain variable region shown in SEQ ID NO: 19 and 33; 20 and 34; 21 and 35; 23 and 37; 24 and 37; 25 and 37; 26 and 37; 27 and 37; 28 and 37; 29 and 37; 30 and 37; 23 and 38; 24 and 38; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 23 and 39; or 23 and 40. In some embodiments, this disclosure provides isolated antibodies that cross-competitively bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) with antibodies comprising the amino acid sequences of the heavy and light chain variable regions shown in SEQ ID NO: 24 and 37, respectively. In some embodiments, this disclosure provides isolated antibodies that cross-competitively bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73) with antibodies that respectively contain the amino acid sequences of the heavy and light chain variable regions shown in SEQ ID NO:27 and 37.

[0759] In some embodiments, this disclosure provides isolated antibodies that bind to the same or overlapping CD73 epitopes (e.g., human, mouse, or cynomolgus monkey CD73 epitopes) as the antibodies described herein, for example, antibodies comprising the heavy and light chain variable region amino acid sequences shown in SEQ ID NO: 19 and 33; 20 and 34; 21 and 35; 23 and 37; 24 and 37; 25 and 37; 26 and 37; 27 and 37; 28 and 37; 29 and 37; 30 and 37; 23 and 38; 24 and 38; 25 and 38; 26 and 38; 27 and 38; 28 and 38; 23 and 39; or 23 and 40, respectively. In some embodiments, this disclosure provides isolated antibodies that bind to the same or overlapping CD73 epitopes (e.g., human, mouse, or cynomolgus monkey CD73 epitopes) as the antibodies described herein, for example, antibodies comprising the heavy and light chain variable region amino acid sequences shown in SEQ ID NO:24 and 37, respectively. In some embodiments, this disclosure provides isolated antibodies that bind to the same or overlapping CD73 epitopes (e.g., human, mouse, or cynomolgus monkey CD73 epitopes) as the antibodies described herein, for example, antibodies comprising the heavy and light chain variable region amino acid sequences shown in SEQ ID NO:27 and 37, respectively. In some embodiments, the epitopes of the antibody can be determined by, for example, NMR spectroscopy, surface plasmon resonance, etc. X-ray diffraction crystallography, ELISA assays, hydrogen / deuterium exchange coupled with mass spectrometry (e.g., liquid chromatography-electrospray mass spectrometry), array-based oligopeptide scanning assays, and / or mutagenesis localization (e.g., site-directed mutagenesis localization). For X-ray crystallography, crystallization can be performed using any method known in the art (e.g., Giegé R et al., (1994) Acta Crystallogr D Biol Crystallogr 50(Pt 4):339-350; McPherson A (1990) Eur J Biochem 189:1-23; Chayen NE (1997) Structure 5:1269-1274; McPherson A (1976) J Biol Chem 251:6300-6303, all of which are incorporated herein by reference in their entirety).

[0760] Antibody: Antigen crystals can be studied using well-known X-ray diffraction techniques and refined using computer software such as X-PLOR (Yale University, 1992, Molecular Simulations, Inc.; see, for example, Meth Enzymol (1985) volumes 114 & 115, eds Wyckoff HW et al.; U.S. Patent Application No. 2004 / 0014194) and BUSTER (Bricogne G (1993) Acta Crystallogr D Biol Crystallogr 49 (Pt 1): 37-60; Bricogne G (1997) Meth Enzymol 276A: 361-423, ed Carter CW; Roversi P et al., (2000) Acta Crystallogr D Biol Crystallogr 56 (Pt 10): 1316-1323, all of which are incorporated herein by reference in their entirety). Mutagenic localization studies can be performed using any method known to those skilled in the art. For descriptions of mutagenic techniques (including alanine scan mutagenesis), see, for example, Champe M et al., (1995) ibid. and Cunningham BC & Wells JA (1989) ibid. In one specific embodiment, alanine scan mutagenesis is used to determine the epitope of the antibody. Alternatively, conventional techniques such as immunoassays can be used to identify antibodies that recognize and bind to the same or overlapping CD73 (e.g., human, mouse, or cynomolgus monkey CD73) epitopes, for example by demonstrating the ability of one antibody to block the binding of another antibody to the target antigen, i.e., competitive binding assays. Competitive binding assays can also be used to determine whether two antibodies have similar binding specificity for an epitope. Competitive binding can be determined in assays where the immunoglobulin being tested inhibits the specific binding of a reference antibody to a common antigen such as CD73 (e.g., human, mouse, or cynomolgus monkey CD73).Many types of competitive binding assays are known, such as: solid-phase direct or indirect radioimmunoassay (RIA), solid-phase direct or indirect enzyme immunoassay (EIA), sandwich competitive assay (see Stahli C et al., (1983) Methods Enzymol 9:242-253); solid-phase direct biotin-avidin EIA (see Kirkland TN et al., (1986) J Immunol 137:3614-9); solid-phase direct labeling assay, solid-phase direct labeling sandwich assay (see Harlow E & Lane D, (1988) Antibodies: A Laboratory Manual, Cold Spring Harbor Press); solid-phase direct labeling RIA using I-125 label (see Morel G et al., (1988) Mol Immunol 25(1):7-15); solid-phase direct biotin-avidin EIA (see Cheung RC et al.). al. (1990) Virology 176:546-52); and directly labeled RIAs (see Moldenhauer, Get al. (1990) Scand J Immunol 32:77-82), all of which are incorporated herein by reference in their entirety. Typically, such assays involve the use of purified antigens (e.g., CD73, such as human, mouse, or cynomolgus monkey CD73) or cells carrying either, bound to a solid, unlabeled test immunoglobulin, and labeled reference immunoglobulin. Competitive inhibition can be measured by determining the amount of labeling bound to a solid surface or cell in the presence of the test immunoglobulin. Typically, an excess of the test immunoglobulin is present. Typically, when an excess of the competitive antibody is present, it will inhibit the specific binding of the reference antibody to the common antigen by at least 50-55%, 55-60%, 60-65%, 65-70%, 70-75%, or more. Competitive binding assays can be configured in a large number of different forms using labeled antigens or labeled antibodies. In the standard form of this assay, the antigen is immobilized on a 96-well plate. Radioactive or enzyme labeling is then used to measure the ability of the unlabeled antibody to block the binding of the labeled antibody to the antigen.For further details, see, for example, Wagener C et al., (1983) J Immunol 130:2308-2315; Wagener C et al., (1984) J Immunol Methods 68:269-274; Kuroki M et al., (1990) Cancer Res 50:4872-4879; Kuroki M et al., (1992) Immunol Invest 21:523-538; Kuroki M et al., (1992) Hybridoma 11:391-407 and Antibodies: A Laboratory Manual, Ed Harlow E & Lane D editors supra, pp.386-389, all of which are incorporated herein by reference in their entirety.

[0761] Any immunoglobulin (Ig) constant region can be used in the antibodies disclosed herein. In some embodiments, the Ig region is a human IgG, IgE, IgM, IgD, IgA, or IgY immunoglobulin molecule, any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or any subclass (e.g., IgG). 2a and IgG 2b ) immunoglobulin molecules.

[0762] In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), the antibodies comprising a heavy chain constant region containing the amino acid sequence of SEQ ID NO: 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, or 58. In some embodiments, this disclosure provides isolated antibodies that specifically bind to CD73 (e.g., human, mouse, or cynomolgus monkey CD73), the antibodies comprising a light chain constant region containing the amino acid sequence of SEQ ID NO: 89 or 93.

[0763] In some embodiments, one, two or more mutations (e.g., amino acid substitutions) are introduced into the Fc region (e.g., the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region (as numbered according to the EU numbering system) of the antibody described herein to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding and / or antigen-dependent cytotoxicity.

[0764] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the hinge region of the Fc region (CH1 domain) such that the number of cysteine ​​residues in the hinge region is altered (e.g., increased or decreased), as described, for example, in U.S. Patent No. 5,677,425, which is incorporated herein by reference in its entirety. The number of cysteine ​​residues in the hinge region of the CH1 domain can be altered to, for example, promote the assembly of light and heavy chains or alter (e.g., increase or decrease) the stability of the antibody.

[0765] In one specific embodiment, one, two, or more mutations (e.g., substitution, insertion, or deletion) are introduced into the constant domain of IgG or its FcRn-binding fragment (preferably an Fc or hinge-Fc domain fragment) to alter (e.g., reduce or increase) the half-life of the antibody in vivo. See, for example, International Publications WO 02 / 060919; WO 98 / 23289; and WO97 / 34631; and U.S. Patents 5,869,046, 6,121,022, 6,277,375, and 6,165,745, all of which are incorporated herein by reference in their entirety, for example, mutations that alter (e.g., reduce or increase) the half-life of the antibody in vivo. In some embodiments, one, two, or more amino acid mutations (e.g., substitution, insertion, or deletion) are introduced into the constant domain of IgG or its FcRn-binding fragment (preferably an Fc or hinge-Fc domain fragment) to reduce the half-life of the antibody in vivo. In other embodiments, one, two, or more amino acid mutations (e.g., substitutions, insertions, or deletions) are introduced into the constant domain of IgG or its FcRn-binding fragment (preferably an Fc or hinge-Fc domain fragment) to increase the antibody's half-life in vivo. In one specific embodiment, the antibody may have one or more amino acid mutations (e.g., substitutions) in a second constant (CH2) domain (residues 231-340 of human IgG1) and / or a third constant (CH3) domain (residues 341-447 of human IgG1) (according to EU numbering system designations). In one specific embodiment, the constant region of IgG1 of the antibody described herein comprises a methionine (M) to tyrosine (Y) substitution at position 252, a serine (S) to threonine (T) substitution at position 254, and a threonine (T) to glutamate (E) substitution at position 256 (according to EU numbering system designations). See U.S. Patent No. 7,658,921, which is incorporated herein by reference in its entirety. This type of mutant IgG (referred to as the “YTE mutant”) has been shown to exhibit a fourfold increase in half-life compared to the wild-type form of the same antibody (see Dall'Acqua WF et al., (2006) JBiol Chem 281:23514-24, which is incorporated herein by reference in its entirety). In some embodiments, the antibody comprises a constant IgG domain (numbered according to the EU numbering system) containing one, two, three, or more amino acid substitutions at positions 251-257, 285-290, 308-314, 385-389, and 428-436.

[0766] In some embodiments, one, two, or more mutations (e.g., amino acid substitutions) are introduced into the Fc region of the antibody described herein (e.g., the CH2 domain (residues 231-340 of human IgG1) and / or the CH3 domain (residues 341-447 of human IgG1) and / or the hinge region (as numbered according to the EU numbering system) to increase or decrease the antibody's affinity for an Fc receptor (e.g., an activated Fc receptor) on the surface of an effector cell. Mutations in the Fc region of antibodies that decrease or increase the antibody's affinity for an Fc receptor, and techniques for introducing such mutations into the Fc receptor or fragments thereof, are known to those skilled in the art. Examples of mutations in the Fc receptor of antibodies that can alter the antibody's affinity for the Fc receptor are described, for example, in Smith P et al., (2012) PNAS 109:6181-6186, U.S. Patent No. 6,737,056, and International Publications WO 02 / 060919; WO 98 / 23289; and WO In 97 / 34631, the entirety of it is incorporated herein by reference.

[0767] In some embodiments, the antibody comprises a variant of the wild-type heavy chain constant region, wherein the variant heavy chain constant region binds FcγRIIB with a higher affinity than the wild-type heavy chain constant region. In some embodiments, the variant heavy chain constant region is a variant human heavy chain constant region, such as the variant human IgG1, variant human IgG2, or variant human IgG4 heavy chain constant region. In some embodiments, the variant human IgG heavy chain constant region comprises one or more of the following amino acid mutations (according to the EU numbering system): G236D, P238D, S239D, S267E, L328F, and L328E. In some embodiments, the constant region of the variant human IgG heavy chain contains a group of amino acid mutations selected from the group consisting of: S267E and L328F; P238D and L328E; P238D and one or more substitutions selected from the group consisting of: E233D, G237D, H268D, P271G and A330R; P238D, E233D, G237D, H268D, P271G and A330R; G236D and S267E; S239D and S267E; V262E, S267E and L328F; and V264E, S267E and L328F (according to the EU numbering system). In some embodiments, FcyRIIB is expressed on cells selected from the group consisting of: macrophages, monocytes, B cells, dendritic cells, endothelial cells and activated T cells.

[0768] In a further embodiment, one, two, or more amino acid substitutions are introduced into the Fc region of the IgG constant domain to alter the effector function of the antibody. For example, one or more amino acids selected from amino acid residues 234, 235, 236, 237, 239, 243, 267, 292, 297, 300, 318, 320, 322, 328, 330, 332, and 396 (according to the EU numbering system) can be substituted with different amino acid residues to give the antibody an altered affinity for the effector ligand, while retaining the antigen-binding ability of the parent antibody. The effector ligand altered to it can be, for example, an Fc receptor or the C1 component of complement. This method is described in more detail in U.S. Patent Nos. 5,624,821 and 5,648,260, each of which is incorporated herein by reference in its entirety. In some embodiments, deletion or inactivation of the constant domain (through point mutation or other means) can reduce Fc receptor binding of circulating antibodies, thereby increasing tumor localization. For descriptions of mutations that result in the loss or deactivation of constant domains to enhance tumor localization, see, for example, U.S. Patent Nos. 5,585,097 and 8,591,886, each of which is incorporated herein by reference in its entirety. In some embodiments, one or more amino acid substitutions may be introduced into the Fc region of the antibody described herein to remove potential glycosylation sites on the Fc region, which may reduce Fc receptor binding (see, for example, Shields RL et al., (2001) J Biol Chem 276:6591-604, which is incorporated herein by reference in its entirety). In various implementations, one or more of the following mutations may be performed in the constant region of the antibody described herein: N297A substitution; N297Q substitution; L234A substitution; L234F substitution; L235A substitution; L235F substitution; L235V substitution; L237A substitution; S239D substitution; E233P substitution; L234V substitution; L235A substitution; C236 deletion; P238A substitution; S239D substitution; F243L substitution; D265A substitution; S267E substitution; L328F substitution; R292P substitution; Y300L substitution; A327Q substitution; P329A substitution; A332L substitution; I332E substitution; or P396L substitution (as per the EU numbering system).

[0769] In some embodiments, mutations selected from the group consisting of D265A, P329A, and combinations thereof (according to EU numbering system) may be made in the constant region of the antibody described herein. In some embodiments, mutations selected from the group consisting of L235A, L237A, and combinations thereof (according to EU numbering system). In some embodiments, mutations selected from the group consisting of S267E, L328F, and combinations thereof (according to EU numbering system). In some embodiments, mutations selected from the group consisting of S239D, I332E, optionally A330L, and combinations thereof (according to EU numbering system). In some embodiments, mutations selected from the group consisting of L235V, F243L, R292P, Y300L, P396L, and combinations thereof (according to EU numbering system). In some implementations, mutations selected from the group consisting of S267E, L328F, and combinations thereof can be made in the constant region of the antibody described herein (as per the EU numbering system).

[0770] In one specific embodiment, the antibody described herein comprises a constant domain of IgG1 having an amino acid substitution of N297Q or N297A (according to EU numbering system numbering). In one embodiment, the antibody described herein comprises a constant domain of IgG1 with a mutation selected from the group consisting of D265A, P329A, and combinations thereof (according to EU numbering system numbering). In another embodiment, the antibody described herein comprises a constant domain of IgG1 with a mutation selected from the group consisting of L234A, L235A, and combinations thereof (according to EU numbering system numbering). In yet another embodiment, the antibody described herein comprises a constant domain of IgG1 with a mutation selected from the group consisting of L234F, L235F, N297A, and combinations thereof (according to EU numbering system numbering). In some embodiments, the amino acid residues (according to EU numbering system numbering) in the constant region of the antibody described herein at positions corresponding to L234, L235, and D265 in the human IgG1 heavy chain are not L, L, and D, respectively. The method is described in detail in International Publication No. WO 14 / 108483, which is incorporated herein by reference in its entirety. In one specific implementation, the amino acids corresponding to positions L234, L235, and D265 in the human IgG1 heavy chain are F, E, and A, respectively; or A, A, and A (as numbered according to the EU numbering system).

[0771] In some embodiments, one or more amino acid residues selected from amino acid residues 329, 331, and 322 (according to EU numbering system) may be replaced with different amino acid residues, thereby giving the antibody altered C1q binding and / or reduced or eliminated complement-dependent cytotoxicity (CDC). This method is described in more detail in U.S. Patent...

Claims

1. An isolated antibody that specifically binds to human CD73, the antibody comprising: a heavy chain variable region comprising complementarity determining regions (CDRs) CDRH1, CDRH2, and CDRH3 and a light chain variable region comprising complementarity determining regions CDRL1, CDRL2, and CDRL3, wherein: (a) CDRH1 comprises an amino acid sequence of X1X2WX3X4 (SEQ ID NO: 13), wherein X1 is S or N; X2 is S or Y; X3 is I or M; and X4 is N or H; (b) CDRH2 comprises the amino acid sequence of X1IYPRX2X3DTNYX4X5KFKX6 (SEQ ID NO: 14), wherein X1 is R or T; X2 is N, A or S; X3 is G or S; X4 is N, A or S; X5 is G or Q; and X6 is D or G; (c) CDRH3 comprises the amino acid sequence of LLDYSMDY (SEQ ID NO: 7); (d) CDRL1 comprises the amino acid sequence of RASQDISX1X2LN (SEQ ID NO: 16), wherein X1 is N or I; and X2 is Y or S; (e) CDRL2 comprises the amino acid sequence of YTSRLHS (SEQ ID NO: 10); and / or (f) CDRL3 comprises the amino acid sequence of QQGNTLPXT (SEQ ID NO: 17), wherein: X is L or W.

2. The isolated antibody of claim 1, wherein the CDRH1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 and 2.

3. The isolated antibody of claim 1 or 2, wherein CDRH2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 3-6. 4 . The isolated antibody of claim 1 , wherein CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 8 and 9. 5 .

5. The isolated antibody of any one of the preceding claims, wherein CDRL3 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 11 and 12.

6. The isolated antibody of any one of the preceding claims, wherein: (a) CDRH1 comprises the amino acid sequence of SSWIN (SEQ ID NO: 2); (b) CDRH2 comprises the amino acid sequence of RIYPRX1GDTNYX2GKFKD (SEQ ID NO: 15), wherein X1 is N, A or S; and X2 is N, A or S; (c) CDRL1 comprises the amino acid sequence of RASQDISX1X2LN (SEQ ID NO: 16), wherein X1 is N or I; and X2 is Y or S; and / or (d) CDRL3 comprises the amino acid sequence of QQGNTLPLT (SEQ ID NO: 12).

7. The isolated antibody of claim 6, wherein CDRH2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 4-6.

8. The isolated antibody of claim 6 or 7, wherein CDRL1 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 8 and 9.

9. The isolated antibody of any one of the preceding claims, wherein CDRH1, CDRH2 and CDRH3 comprise the CDRH1, CDRH2 and CDRH3 amino acid sequences set forth in SEQ ID NO: 2, 4 and 7; 2, 5 and 7; 1, 3 and 7; or 2, 6 and 7, respectively.

10. The isolated antibody of any of the preceding claims, wherein CDRL1, CDRL2 and CDRL3 comprise the CDRL1, CDRL2 and CDRL3 amino acid sequences set forth in SEQ ID NOs: 8, 10 and 12; 9, 10 and 12; or 8, 10 and 11, respectively.

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