Materials and methods for immune effector cell redirection

Abstract

A molecule comprising a first means for associating with or activating natural killer (NK) cells and a second means for binding to tumor cells, said molecule being capable of inducing NK cell-dependent cytotoxicity against tumor cells.

Description

[Technical Field]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims the benefit of U.S. Patent Application Nos. 63 / 168,605, filed March 31, 2021, 63 / 168,611, filed March 31, 2021, 63 / 168,618, filed March 31, 2021, 63 / 168,621, filed March 31, 2021, and 63 / 168,628, filed March 31, 2021, each of which is incorporated by reference in its entirety.

[0002] (Reference to electronically submitted sequence listing) This application contains a Sequence Listing that has been submitted electronically via EFS-Web as an ASCII Sequence Listing with the file name "14620-648-228_SEQ_LISTING", created on March 24, 2022, and having a size of 181,332 bytes. The Sequence Listing submitted via EFS-Web is a part of the present specification and is incorporated herein by reference in its entirety.

[0003] FIELD OF THE INVENTION The present disclosure relates, inter alia, to natural killer cell-engaging factors, including anti-NKG2d molecules, anti-NKp46 molecules, and multispecific molecules comprising them or fragments thereof, as well as nucleic acids and expression vectors encoding the molecules, recombinant cells containing these vectors, and compositions comprising these molecules. Methods of making and using the molecules to redirect immune effector cells against tumor cells are also provided. [Background technology]

[0004] Tumor cells can be therapeutically targeted for destruction by antibodies. Therapeutic antibodies engage immune effector cells and can target tumor cells for destruction using a number of mechanisms. Effector cells can be redirected against tumor cells using bispecific antibodies (bsAb), which bind to tumor cells and effector cells, bringing them into close proximity. Alternatively, monoclonal antibodies (mAb) can engage tumor cells through their variable regions and recruit effector cells through interactions between their Fc regions and Fc g receptors, which are expressed primarily on monocytes, macrophages, and NK cells. Summary of the Invention

[0005] The present inventors have now discovered the first multispecific antibodies described herein that comprise a first binding domain that binds to a first antigen expressed on Natural Killer (NK) cells and a second binding domain that binds to a second antigen expressed on tumor cells.

[0006] In one aspect, provided herein is a multispecific antibody comprising a first binding domain that binds to a first antigen expressed on a natural killer (NK) cell and a second binding domain that binds to a second antigen.

[0007] In some embodiments of the multispecific antibodies provided herein, the first antigen is an NK cell activating receptor.

[0008] In some embodiments of the multispecific antibodies provided herein, the first antigen is NKG2d.

[0009] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises: (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 4, a VH CDR2 having the amino acid sequence of SEQ ID NO: 5, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 6; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 10, a VH CDR2 having the amino acid sequence of SEQ ID NO: 11, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 12; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 16, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 18; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 22, a VH CDR2 having the amino acid sequence of SEQ ID NO: 23, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 24. a heavy chain variable region (VH) comprising (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 7, a VL CDR2 having the amino acid sequence of SEQ ID NO: 8, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 9, or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 28, a VL CDR2 having the amino acid sequence of SEQ ID NO: 29, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 30, and (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 13, a VL CDR2 having the amino acid sequence of SEQ ID NO: 14, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 15, (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 19, a VL CDR2 having the amino acid sequence of SEQ ID NO: 20, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 21, or (d) a VL CDR1 having the amino acid sequence of SEQ ID NO: 25, a VL CDR2 having the amino acid sequence of SEQ ID NO: 26, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 27. and (e) a light chain variable region (VL) comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 31, a VL CDR2 having the amino acid sequence of SEQ ID NO: 32, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 33.

[0010] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO:2 and a VL having the amino acid sequence of SEQ ID NO:3.

[0011] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises: (i) (a) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 36, a VH CDR2 having the amino acid sequence of SEQ ID NO: 37, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 38; (b) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 42, a VH CDR2 having the amino acid sequence of SEQ ID NO: 43, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 44; (c) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 48, a VH CDR2 having the amino acid sequence of SEQ ID NO: 49, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 50; (d) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 54, a VH CDR2 having the amino acid sequence of SEQ ID NO: 55, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 56. and (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 51, a VL CDR2 having the amino acid sequence of SEQ ID NO: 52, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 53; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 60, a VL CDR2 having the amino acid sequence of SEQ ID NO: 61, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 62; and (ii) a heavy chain variable region (VH) comprising: (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 39, a VL CDR2 having the amino acid sequence of SEQ ID NO: 40, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 41; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 45, a VL CDR2 having the amino acid sequence of SEQ ID NO: 46, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 47; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 51, a VL CDR2 having the amino acid sequence of SEQ ID NO: 52, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 53; and (e) a light chain variable region (VL) comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 63, a VL CDR2 having the amino acid sequence of SEQ ID NO: 64, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 65.

[0012] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO: 35.

[0013] In some embodiments of the multispecific antibodies provided herein, the first antigen is NKp46.

[0014] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises: (i) (a) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 69, a VH CDR2 having the amino acid sequence of SEQ ID NO: 70, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 71; (b) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 75, a VH CDR2 having the amino acid sequence of SEQ ID NO: 76, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 77; (c) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 81, a VH CDR2 having the amino acid sequence of SEQ ID NO: 82, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 83; (d) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 87, a VH CDR2 having the amino acid sequence of SEQ ID NO: 88, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 89. a heavy chain variable region (VH) comprising (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 72, a VL CDR2 having the amino acid sequence of SEQ ID NO: 73, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 74; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 78, a VL CDR2 having the amino acid sequence of SEQ ID NO: 79, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 80; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 84, a VL CDR2 having the amino acid sequence of SEQ ID NO: 85, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 86; or (d) a VL CDR1 having the amino acid sequence of SEQ ID NO: 90, a VL CDR2 having the amino acid sequence of SEQ ID NO: 91, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 92. and (e) a light chain variable region (VL) comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 96, a VL CDR2 having the amino acid sequence of SEQ ID NO: 97, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 98.

[0015] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 67 and a VL having the amino acid sequence of SEQ ID NO: 68.

[0016] In some embodiments of the multispecific antibodies provided herein, the second antigen is on a cell surface.

[0017] In some embodiments of the multispecific antibodies provided herein, the second antigen is expressed on a tumor cell.

[0018] In some embodiments of the multispecific antibodies provided herein, the second antigen is a tumor specific antigen (TSA) or a tumor associated antigen (TAA).

[0019] In some embodiments of the multispecific antibodies provided herein, the second antigen is BCMA.

[0020] In some embodiments of the multispecific antibodies provided herein, the second antigen is GPRC5d.

[0021] In some embodiments of the multispecific antibodies provided herein, the first binding domain is humanized. In some embodiments of the multispecific antibodies provided herein, the second binding domain is humanized. In some embodiments of the multispecific antibodies provided herein, both the first binding domain and the second binding domain are humanized.

[0022] In some embodiments of the multispecific antibodies provided herein, the multispecific antibody is an IgG antibody.

[0023] In some embodiments of the multispecific antibodies provided herein, the IgG antibody is an IgG1, IgG2, IgG3, or IgG4 antibody.

[0024] In some embodiments of the multispecific antibodies provided herein, the IgG antibody is an IgG1 antibody.

[0025] In some embodiments of the multispecific antibodies provided herein, the IgG1 comprises a silent mutation. In some embodiments of the multispecific antibodies provided herein, the IgG1 comprises an AAS mutation. In some embodiments, the multispecific antibodies comprising an AAS mutation can induce NK cell-dependent cytotoxicity of tumor cells.

[0026] In some embodiments of the multispecific antibodies provided herein, the IgG1 comprises mutations to enhance the effector function of the antibody. In some embodiments of the multispecific antibodies provided herein, the IgG1 comprises K248E / T437R mutations. In some embodiments, multispecific antibodies comprising the K248E / T437R mutations lack anti-NK cell cytotoxicity.

[0027] In some embodiments of the multispecific antibodies provided herein, the Fc region is defucosylated.

[0028] In some embodiments of the multispecific antibodies provided herein, the multispecific antibody is a bispecific antibody.

[0029] In some embodiments of the multispecific antibodies provided herein, the bispecific antibody is a bipod scaffold structure.

[0030] In some embodiments of the multispecific antibodies provided herein, the first binding domain is a Fab region and the second binding domain is an scFv region.

[0031] In some embodiments of the multispecific antibodies provided herein, the bispecific antibody is a Morrison scaffold.

[0032] In some embodiments of the multispecific antibodies provided herein, the first binding domain comprises two Fab regions and the second binding domain comprises two scFv regions.

[0033] In some embodiments of the multispecific antibodies provided herein, the multispecific antibodies lack anti-NK cell cytotoxicity.

[0034] In some embodiments of the multispecific antibodies provided herein, the multispecific antibodies induce NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 500 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 300 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 100 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 50 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 20 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 15 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 10 pM. 50 Guide with.

[0035] In some embodiments of the multispecific antibodies provided herein, IC 50 is assessed using a mixture of NK effector cells and target cells expressing a second antigen.

[0036] In some embodiments of the multispecific antibodies provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 10:1. In some embodiments of the multispecific antibodies provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 5:1. In some embodiments, the ratio of effector cells to target cells is about 0.1:1 to about 2:1. In some embodiments, the ratio of effector cells to target cells is about 1:1.

[0037] In another aspect, nucleic acids encoding the multispecific antibodies provided herein are provided. Also provided are vectors comprising nucleic acids encoding the multispecific antibodies provided herein. Also provided are host cells comprising vectors comprising nucleic acids encoding the multispecific antibodies provided herein. Also provided are kits comprising vectors comprising nucleic acids encoding the multispecific antibodies provided herein and packaging therefor.

[0038] In another aspect, provided herein is an antibody that binds to NKG2d, comprising: (i) (a) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 4, a VH CDR2 having the amino acid sequence of SEQ ID NO: 5, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 6; (b) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 10, a VH CDR2 having the amino acid sequence of SEQ ID NO: 11, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 12; (c) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 16, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 18; (d) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 22, a VH CDR2 having the amino acid sequence of SEQ ID NO: 23, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 24. a heavy chain variable region (VH) comprising (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 7, a VL CDR2 having the amino acid sequence of SEQ ID NO: 8, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 9, (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 13, a VL CDR2 having the amino acid sequence of SEQ ID NO: 14, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 15, (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 19, a VL CDR2 having the amino acid sequence of SEQ ID NO: 20, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 21, or (d) a VL CDR1 having the amino acid sequence of SEQ ID NO: 25, a VL CDR2 having the amino acid sequence of SEQ ID NO: 26, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 27, and (e) a light chain variable region (VL) comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 31, a VL CDR2 having the amino acid sequence of SEQ ID NO: 32, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 33.

[0039] In some embodiments of the antibodies that bind to NKG2d provided herein, the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO:2 and a VL having the amino acid sequence of SEQ ID NO:3.

[0040] In another aspect, provided herein is an antibody that binds to NKG2d, comprising: (i) (a) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 36, a VH CDR2 having the amino acid sequence of SEQ ID NO: 37, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 38; (b) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 42, a VH CDR2 having the amino acid sequence of SEQ ID NO: 43, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 44; (c) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 48, a VH CDR2 having the amino acid sequence of SEQ ID NO: 49, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 50; (d) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 54, a VH CDR2 having the amino acid sequence of SEQ ID NO: 55, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 56 and (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 51, a VL CDR2 having the amino acid sequence of SEQ ID NO: 52, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 53; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 60, a VL CDR2 having the amino acid sequence of SEQ ID NO: 61, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 62; and (ii) a heavy chain variable region (VH) comprising: (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 39, a VL CDR2 having the amino acid sequence of SEQ ID NO: 40, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 41; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 45, a VL CDR2 having the amino acid sequence of SEQ ID NO: 46, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 47; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 51, a VL CDR2 having the amino acid sequence of SEQ ID NO: 52, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 53; and (e) a light chain variable region (VL) comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 63, a VL CDR2 having the amino acid sequence of SEQ ID NO: 64, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 65.

[0041] In some embodiments of the antibodies that bind to NKG2d provided herein, the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO: 35.

[0042] In another aspect, provided herein is an antibody that binds to NKp46, comprising: (i) (a) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 69, a VH CDR2 having the amino acid sequence of SEQ ID NO: 70, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 71; (b) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 75, a VH CDR2 having the amino acid sequence of SEQ ID NO: 76, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 77; (c) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 81, a VH CDR2 having the amino acid sequence of SEQ ID NO: 82, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 83; (d) a VH complementarity determining region (CDR)1 having the amino acid sequence of SEQ ID NO: 87, a VH CDR2 having the amino acid sequence of SEQ ID NO: 88, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 89 a heavy chain variable region (VH) comprising (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 72, a VL CDR2 having the amino acid sequence of SEQ ID NO: 73, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 74; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 78, a VL CDR2 having the amino acid sequence of SEQ ID NO: 79, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 80; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 84, a VL CDR2 having the amino acid sequence of SEQ ID NO: 85, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 86; or (d) a VL CDR1 having the amino acid sequence of SEQ ID NO: 90, a VL CDR2 having the amino acid sequence of SEQ ID NO: 91, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 92. and (e) a light chain variable region (VL) comprising a VL CDR1 having the amino acid sequence of SEQ ID NO: 96, a VL CDR2 having the amino acid sequence of SEQ ID NO: 97, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 98.

[0043] In some embodiments of the antibodies that bind to NKp46 provided herein, the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 67 and a VL having the amino acid sequence of SEQ ID NO: 68.

[0044] In another aspect, provided herein are nucleic acids encoding the antibodies provided herein. Also provided are vectors comprising nucleic acids encoding the antibodies provided herein. Also provided are host cells comprising vectors comprising nucleic acids encoding the antibodies provided herein. Also provided are kits comprising vectors comprising nucleic acids encoding the antibodies provided herein and packaging therefor.

[0045] In yet another aspect, provided herein is a pharmaceutical composition comprising a multispecific antibody provided herein and a pharmaceutically acceptable carrier, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen expressed on natural killer (NK) cells and a second binding domain that binds to a second antigen.

[0046] In some embodiments of the pharmaceutical compositions provided herein, the first antigen is an NK cell activating receptor.

[0047] In some embodiments of the pharmaceutical compositions provided herein, the first antigen is NKG2d.

[0048] In some embodiments of the pharmaceutical compositions provided herein, the first antigen is NKp46.

[0049] In some embodiments of the pharmaceutical compositions provided herein, the second antigen is on the cell surface.

[0050] In some embodiments of the pharmaceutical compositions provided herein, the second antigen is expressed on a tumor cell.

[0051] In some embodiments of the pharmaceutical compositions provided herein, the second antigen is a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA).

[0052] In some embodiments of the pharmaceutical compositions provided herein, the second antigen is BCMA.

[0053] In some embodiments of the pharmaceutical compositions provided herein, the second antigen is GPRC5d.

[0054] In some embodiments of the pharmaceutical compositions provided herein, the first binding domain is humanized. In some embodiments of the pharmaceutical compositions provided herein, the second binding domain is humanized. In some embodiments of the pharmaceutical compositions provided herein, both the first binding domain and the second binding domain are humanized.

[0055] In some embodiments of the pharmaceutical compositions provided herein, the multispecific antibody is an IgG antibody.

[0056] In some embodiments of the pharmaceutical compositions provided herein, the IgG antibody is an IgG1, IgG2, IgG3, or IgG4 antibody.

[0057] In some embodiments of the pharmaceutical compositions provided herein, the IgG antibody is an IgG1 antibody.

[0058] In some embodiments of the pharmaceutical compositions provided herein, the IgG1 comprises a silent mutation. In some embodiments of the pharmaceutical compositions provided herein, the IgG1 comprises an AAS mutation.

[0059] In some embodiments of the pharmaceutical compositions provided herein, the IgG1 comprises mutations to enhance the effector function of the antibody. In some embodiments of the pharmaceutical compositions provided herein, the IgG1 comprises K248E / T437R mutations.

[0060] In some embodiments of the pharmaceutical compositions provided herein, the Fc region is defucosylated.

[0061] In some embodiments of the pharmaceutical compositions provided herein, the multispecific antibody is a bispecific antibody.

[0062] In some embodiments of the pharmaceutical compositions provided herein, the bispecific antibody is a bipod scaffold.

[0063] In some embodiments of the pharmaceutical compositions provided herein, the first binding domain is a Fab region and the second binding domain is an scFv region.

[0064] In some embodiments of the pharmaceutical compositions provided herein, the bispecific antibody is a Morrison scaffold.

[0065] In some embodiments of the pharmaceutical compositions provided herein, the first binding domain comprises two Fab regions and the second binding domain comprises two scFv regions.

[0066] In some embodiments of the pharmaceutical compositions provided herein, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 500 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 300 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 100 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 50 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 20 pM. 50In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 15 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 10 pM. 50 Guide with.

[0067] In some embodiments of the pharmaceutical compositions provided herein, IC 50 is assessed using a mixture of NK effector cells and target cells expressing a second antigen.

[0068] In some embodiments of the pharmaceutical compositions provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 10:1. In some embodiments of the pharmaceutical compositions provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 5:1. In some embodiments, the ratio of effector cells to target cells is about 0.1:1 to about 2:1. In some embodiments, the ratio of effector cells to target cells is about 1:1.

[0069] In yet another aspect, provided herein is a process for making a multispecific antibody, the process comprising introducing into a host cell one or more nucleic acids encoding a first binding domain that binds to a first antigen expressed on an NK cell and a second binding domain that binds to a second antigen.

[0070] In some embodiments of the processes for making multispecific antibodies provided herein, the first antigen is an NK cell activating receptor.

[0071] In some embodiments of the processes for making multispecific antibodies provided herein, the multispecific antibody is a multispecific antibody provided herein.

[0072] In some embodiments of the processes for making multispecific antibodies provided herein, the first antigen is NKp46. In some embodiments of the processes for making multispecific antibodies provided herein, the first antigen is NKG2d.

[0073] In some embodiments of the processes for making multispecific antibodies provided herein, the second antigen is on the cell surface.

[0074] In some embodiments of the processes for making multispecific antibodies provided herein, the second antigen is expressed on a tumor cell.

[0075] In some embodiments of the processes for making multispecific antibodies provided herein, the second antigen is a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA).

[0076] In some embodiments of the processes for making multispecific antibodies provided herein, the second antigen is BCMA.

[0077] In some embodiments of the processes for making multispecific antibodies provided herein, the second antigen is GPRC5d.

[0078] In some embodiments of the processes for making multispecific antibodies provided herein, the first binding domain is humanized. In some embodiments of the processes for making multispecific antibodies provided herein, the second binding domain is humanized. In some embodiments of the processes for making multispecific antibodies provided herein, both the first binding domain and the second binding domain are humanized.

[0079] In some embodiments of the processes for making multispecific antibodies provided herein, the multispecific antibody is an IgG antibody.

[0080] In some embodiments of the processes for making multispecific antibodies provided herein, the IgG antibody is an IgG1, IgG2, IgG3, or IgG4 antibody.

[0081] In some embodiments of the processes for making multispecific antibodies provided herein, the IgG antibody is an IgG1 antibody.

[0082] In some embodiments of the processes provided herein, the IgG1 comprises a silent mutation.In some embodiments of the processes provided herein, the IgG1 comprises an AAS mutation.

[0083] In some embodiments of the processes provided herein, the IgG1 comprises mutations to enhance the effector function of the antibody. In some embodiments of the processes provided herein, the IgG1 comprises K248E / T437R mutations.

[0084] In some embodiments of the processes provided herein, the Fc region is defucosylated.

[0085] In some embodiments of the processes for making multispecific antibodies provided herein, the multispecific antibody is a bispecific antibody.

[0086] In some embodiments of the processes for making multispecific antibodies provided herein, the bispecific antibody is a Bipod scaffold structure.

[0087] In some embodiments of the processes for making multispecific antibodies provided herein, the first binding domain is a Fab region and the second binding domain is an scFv region.

[0088] In some embodiments of the processes for making multispecific antibodies provided herein, the bispecific antibody is a Morrison scaffold.

[0089] In some embodiments of the processes for making multispecific antibodies provided herein, the first binding domain comprises two Fab regions and the second binding domain comprises two scFv regions.

[0090] In some embodiments of the processes for making multispecific antibodies provided herein, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 500 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 300 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 100 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 50 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 20 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 15 pM. 50 In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC of less than about 10 pM. 50 Guide with.

[0091] In some embodiments of the processes for making multispecific antibodies provided herein, IC 50 is assessed using a mixture of NK effector cells and target cells expressing a second antigen.

[0092] In some embodiments of the processes for producing multispecific antibodies provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 10:1. In some embodiments of the processes for producing multispecific antibodies provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 5:1. In some embodiments, the ratio of effector cells to target cells is about 0.1:1 to about 2:1. In some embodiments, the ratio of effector cells to target cells is about 1:1.

[0093] In yet another aspect, provided herein is a method of directing NK cells to target cells, comprising contacting the NK cells with a multispecific antibody, thereby directing the NK cells to the target cells, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on the NK cell and a second binding domain that binds to a second antigen on the target cell.

[0094] In another aspect, provided herein is a use of a multispecific antibody for directing NK cells to target cells, comprising contacting NK cells with the multispecific antibody, thereby directing the NK cells to the target cells, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on the NK cell and a second binding domain that binds to a second antigen on the target cell.

[0095] In another aspect, provided herein is a method of activating an NK cell, comprising contacting the NK with a multispecific antibody, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on the NK cell and a second binding domain that binds to a second antigen on a target cell.

[0096] In another aspect, provided herein is a use of a multispecific antibody for activating NK cells, comprising contacting the NK with a multispecific antibody, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on the NK cell and a second binding domain that binds to a second antigen on the target cell.

[0097] In another aspect, provided herein is a method of inhibiting the growth or proliferation of a target cell that expresses a second antigen on its cell surface, comprising contacting the target cell with a multispecific antibody, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on an NK cell and a second binding domain that binds to a second antigen.

[0098] In another aspect, provided herein is a use of a multispecific antibody for inhibiting the growth or proliferation of a target cell that expresses a second antigen on the cell surface, wherein the use of the multispecific antibody comprises contacting the target cell with the multispecific antibody, and wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on an NK cell and a second binding domain that binds to a second antigen.

[0099] In another aspect, provided herein is a method of ablating target cells expressing a second antigen in a subject, or treating a disease or disorder caused in whole or in part by target cells expressing a second antigen, comprising administering to the subject an effective amount of a multispecific antibody, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on an NK cell and a second binding domain that binds to the second antigen.

[0100] In another aspect, provided herein is a method of eliminating target cells expressing a second antigen in a subject, or use of a multispecific antibody for treating a disease or disorder caused in whole or in part by target cells expressing a second antigen, comprising administering to the subject an effective amount of a multispecific antibody, wherein the multispecific antibody comprises a first binding domain that binds to a first antigen on an NK cell and a second binding domain that binds to the second antigen.

[0101] In some embodiments, the subject is a subject in need of such treatment. In some embodiments, the subject is a human.

[0102] In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer is a solid tumor cancer.

[0103] In some embodiments of the methods provided herein, the first antigen is an NK cell activating receptor.

[0104] In some embodiments of the methods provided herein, the first antigen is NKG2d. In some embodiments of the methods provided herein, the first antigen is NKp46. In some embodiments, the multispecific antibody is a multispecific antibody provided herein.

[0105] In some embodiments of the methods provided herein, the second antigen is on the cell surface.

[0106] In some embodiments of the methods provided herein, the second antigen is expressed on a tumor cell.

[0107] In some embodiments of the methods provided herein, the second antigen is a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA).

[0108] In some embodiments of the methods provided herein, the second antigen is BCMA.

[0109] In some embodiments of the methods provided herein, the second antigen is GPRC5d.

[0110] In some embodiments of the methods provided herein, the first binding domain is humanized. In some embodiments of the methods provided herein, the second binding domain is humanized. In some embodiments of the methods provided herein, both the first binding domain and the second binding domain are humanized.

[0111] In some embodiments of the methods provided herein, the multispecific antibody is an IgG antibody.

[0112] In some embodiments of the methods provided herein, the IgG antibody is an IgG1, IgG2, IgG3, or IgG4 antibody.

[0113] In some embodiments of the methods provided herein, the IgG antibody is an IgG1 antibody.

[0114] In some embodiments of the methods provided herein, the IgG1 comprises a silent mutation.In some embodiments of the methods provided herein, the IgG1 comprises an AAS mutation.

[0115] In some embodiments of the methods provided herein, the IgG1 comprises mutations to enhance the effector function of the antibody. In some embodiments of the methods provided herein, the IgG1 comprises K248E / T437R mutations.

[0116] In some embodiments of the methods provided herein, the Fc region is defucosylated.

[0117] In some embodiments of the methods provided herein, the multispecific antibody is a bispecific antibody.

[0118] In some embodiments of the methods provided herein, the bispecific antibody is a bipod scaffold.

[0119] In some embodiments of the methods provided herein, the first binding domain is a Fab region and the second binding domain is an scFv region.

[0120] In some embodiments of the methods provided herein, the bispecific antibody is a Morrison scaffold.

[0121] In some embodiments of the methods provided herein, the first binding domain comprises two Fab regions and the second binding domain comprises two scFv regions.

[0122] In some embodiments of the methods provided herein, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 500 pM. In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 300 pM. In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 100 pM. In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 50 pM. In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 20 pM. In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 15 pM. In some embodiments, the multispecific antibody induces NK cell-dependent cytotoxicity of tumor cells in vitro with an IC50 of less than about 10 pM.

[0123] In some embodiments of the methods provided herein, the IC50 is assessed using a mixture of NK effector cells and target cells expressing a second antigen.

[0124] In some embodiments of the methods provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 10:1. In some embodiments of the methods provided herein, the ratio of effector cells to target cells is about 0.01:1 to about 5:1. In some embodiments, the ratio of effector cells to target cells is about 0.1:1 to about 2:1. In some embodiments, the ratio of effector cells to target cells is about 0.5:1. In some embodiments, the ratio of effector cells to target cells is about 1:1.

[0125] In another aspect, a molecule is provided that comprises a first means for associating with or activating an NK cell and a second means for binding to a tumor cell, wherein the molecule is capable of inducing NK cell-dependent cytotoxicity against the tumor cell.

[0126] In some embodiments of the molecules provided herein, the first means comprises a first binding domain that binds to a first antigen expressed on an NK cell, and the second means comprises a second binding domain that binds to a second antigen expressed on a tumor cell.

[0127] In some embodiments of the molecules provided herein, the first antigen is an NK cell activating receptor.

[0128] In some embodiments of the molecules provided herein, the first antigen is NKG2d.

[0129] In some embodiments of the molecules provided herein, the first antigen is NKp46.

[0130] In some embodiments of the molecules provided herein, the second antigen is a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA).

[0131] In some embodiments of the molecules provided herein, the second antigen is BCMA.

[0132] In some embodiments of the molecules provided herein, the second antigen is GPRC5d.

[0133] In another aspect, a process is provided for generating an antibody that binds to more than one target molecule, the process comprising performing the functions of obtaining a binding domain capable of binding to a first antigen on an NK cell, performing the functions of obtaining a binding domain capable of binding to a second antigen on a tumor cell, and performing the functions of providing a molecule capable of binding to the first antigen and the second antigen.

[0134] In another aspect, provided herein is a method of directing NK cells to target cells, the method comprising contacting NK cells with a molecule provided herein.

[0135] In another aspect, provided herein are methods of activating NK cells, comprising contacting NK cells with a molecule provided herein.

[0136] In another aspect, provided herein is a method of inhibiting the growth or proliferation of a target cell, the method comprising contacting the target cell with a molecule provided herein.

[0137] In another aspect, provided herein is a method of ablating target cells that express a second antigen in a subject, or treating a disease or disorder caused in whole or in part by target cells that express a second antigen, comprising administering an effective amount of a molecule provided herein. [Brief explanation of the drawings]

[0138] [Figure 1] 1 shows the results of serum titers from Omni rats immunized with NKGW1. [Figure 2] 1 shows a bead-based assay for NK cell agonism. [Figure 3] 1 shows the results of serum titers from Omni rats immunized with NKp46. [Figure 4] The structure of bsAb is shown. [Figure 5A] 1 shows the results of an analysis of the ability of BsAbs to mediate NK cell line cytotoxicity. [Figure 5B] 1 shows the results of an analysis of the ability of BsAbs to mediate NK cell line cytotoxicity. [Figure 5C] 1 shows the results of an analysis of the ability of BsAbs to mediate NK cell line cytotoxicity. [Figure 5D] 1 shows the results of an analysis of the ability of BsAbs to mediate NK cell line cytotoxicity. [Figure 6]1 shows the structures of additional BCMA-binding monovalent mAbs and bsAbs. [Figure 7] 1 shows the potentiation of NK cells in ADCC by 72 hour pretreatment with 1 ng / ml TGFb, and the functional rescue of ADCC by incorporation of the NKp46 binding arm in the effector molecule N46B10.AFU. [Figure 8A] 1 shows the enhancement of NK cells in ADCC under hypoxic conditions by incorporation of an NKp46 binding arm in the effector molecule N46B10.AFU compared to BCMB1106.AFU which does not contain an NKp46 binding arm. [Figure 8B] 1 shows the enhancement of NK cells in ADCC under hypoxic conditions by incorporation of an NKp46 binding arm in the effector molecule N46B10.AFU compared to BCMB1106.AFU which does not contain an NKp46 binding arm. [Figure 8C] 1 shows the enhancement of NK cells in ADCC under hypoxic conditions by incorporation of an NKp46 binding arm in the effector molecule N46B10.AFU compared to BCMB1106.AFU which does not contain an NKp46 binding arm. [Figure 9] Figure 1 shows the lack of anti-NK CDC activity by the BCMAxNKp46 bispecific molecule in the presence of human serum, in contrast to the anti-CD38 positive control mAb, which shows potent anti-NK CDC killing. DETAILED DESCRIPTION OF THE INVENTION

[0139] The present disclosure is based in part on novel molecules that bind to antigens on NK cells and multispecific binding molecules comprising same or fragments thereof, and the advanced properties of these novel molecules, e.g., a first means capable of binding to a first antigen present on an NK cell, and a second means capable of binding to a second antigen, e.g., on a tumor cell.

[0140] definition The techniques and procedures described or referenced herein include those generally well understood and / or commonly employed by those skilled in the art using conventional techniques, such as the widely used techniques described in Molecular Cloning: A Laboratory Manual (Sambrook, et al., 3rd ed. 2001), Current Protocols in Molecular Biology (Ausubel, et al. eds., 2003), Therapeutic Monoclonal Antibodies: From Bench to Clinic (An ed. 2009), Monoclonal Antibodies: Methods and Protocols (Albitar, ed. 2010), and Antibody Engineering Vols 1 and 2 (Kontermann and Dubel, eds., 2nd ed. 2010).

[0141] Unless otherwise defined herein, technical and scientific terms used herein have the meanings that are commonly understood by those skilled in the art. For the purposes of interpreting this specification, the following explanations of terms shall apply, and where appropriate, terms used in the singular shall also include the plural and vice versa. In the event that any explanation of a term provided contradicts any document incorporated herein by reference, the explanation of the term provided below shall prevail.

[0142] The terms "antibody," "immunoglobulin," or "Ig" are used interchangeably herein and are used in the broadest sense, specifically encompassing monoclonal antibodies (including agonist, antagonist, neutralizing, full-length, or intact monoclonal antibodies), antibody compositions with polyepitopic or monoepitopic specificity, polyclonal or monovalent antibodies, multivalent antibodies, and multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, so long as they exhibit the desired biological activity, as described below. Antibodies may be human, humanized, chimeric, and / or affinity matured, and may also be antibodies from other species, such as mouse and rabbit. The term "antibody" is intended to include polypeptide products of B cells within the immunoglobulin class of polypeptides that can bind to a specific molecular antigen and are composed of two identical paired polypeptide chains, each pair having one heavy chain (approximately 50-70 kDa) and one light chain (approximately 25 kDa), with the amino-terminal portion of each chain containing a variable region of about 100 to about 130 or more amino acids, and the carboxy-terminal portion of each chain containing a constant region. See, e.g., Antibody Engineering (Borrebaeck, ed., 2nd ed. 1995) and Kuby, Immunology (3rd ed. 1997). In specific embodiments, specific molecular antigens can be bound by antibodies provided herein that contain a polypeptide or epitope. Antibodies also include, but are not limited to, synthetic antibodies, recombinantly produced antibodies, camelized antibodies or humanized variants thereof, intracellular antibodies, and anti-idiotypic (anti-Id) antibodies. As used herein, the term "antibody" also includes any binding molecule having an Fc region and a functional fragment (e.g., an antigen-binding fragment) of any of the above, where a functional fragment refers to a portion of an antibody heavy or light chain polypeptide that retains some or all of the binding activity of the antibody from which the fragment is derived.Non-limiting examples of functional fragments (e.g., antigen-binding fragments) include single-chain Fvs (scFv) (including, e.g., monospecific, bispecific, etc.), Fab fragments, F(ab') fragments, F(ab)2 fragments, F(ab')2 fragments, disulfide-linked Fvs (dsFv), Fd fragments, Fv fragments, diabodies, triabodies, tetrabodies, and minibodies. In particular, antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, e.g., antigen-binding domains or molecules comprising an antigen-binding site (e.g., one or more CDRs of an antibody) that binds to an antigen. Such antibody fragments can be found, for example, in Harlow and Lane, Antibodies: A Laboratory Manual (1989), Mol. Biology and Biotechnology: A Comprehensive Desk Reference (Myers, ed., 1995), Huston, et al., 1993, Cell Biophysics 22:189-224, Pluckthun and Skerra, 1989, Meth. Enzymol. 178:497-515, and Day, Advanced Immunochemistry (2d ed. 1990). The antibodies provided herein may be of any class (e.g., IgG, IgE, IgM, IgD, and IgA) or any subclass (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2) of immunoglobulin molecules. The antibodies may be agonist or antagonist antibodies.

[0143] An "antigen" is a structure to which an antibody can selectively bind. A target antigen can be a polypeptide, carbohydrate, nucleic acid, lipid, hapten, or other naturally occurring or synthetic compound. In some embodiments, the target antigen is a polypeptide. In certain embodiments, the antigen is associated with a cell, e.g., present on or within a cell.

[0144] An "intact" antibody is one that contains an antigen-binding site as well as a constant domain (CL) and at least heavy chain constant regions CH1, CH2, and CH3. The constant region may include a human constant region or a variant of its amino acid sequence. In certain embodiments, an intact antibody has one or more effector functions.

[0145] The terms "binds" or "binding" refer to interactions between molecules, including, for example, forming a complex. The interactions can be non-covalent interactions, including, for example, hydrogen bonds, ionic bonds, hydrophobic interactions, and / or van der Waals interactions. A complex can also include the binding of two or more molecules held together by covalent or non-covalent bonds, interactions, or forces. The strength of the total non-covalent interactions between a single antigen-binding site of an antibody and a single epitope of a target molecule, such as an antigen, is the affinity of the antibody or functional fragment for that epitope. The dissociation rate (k off ) and association rate (k on ) and the ratio (k off / k on ) is the dissociation constant K D and has an inverse relationship with affinity. D The lower the value, the higher the affinity of the antibody. D The value of k varies for different complexes of antibody and antigen. on and k off The dissociation constant K of the antibodies provided herein depends on both D can be determined using any of the methods provided herein or any other method known to those skilled in the art. The affinity at one binding site does not necessarily reflect the true strength of the interaction between the antibody and the antigen. When a complex antigen containing multiple repeating antigenic determinants, such as a multivalent antigen, comes into contact with an antibody containing multiple binding sites, the interaction of the antibody with the antigen at one site will increase the probability of reaction at a second site. The strength of multiple interactions between such a multivalent antibody and an antigen is called avidity.

[0146] In reference to the antibodies described herein, terms such as "binds to," "specifically binds to," and similar terms are also used interchangeably herein to refer to an antibody of an antigen-binding domain that specifically binds to an antigen, such as a polypeptide. An antibody or antigen-binding domain that binds to an antigen or specifically binds to an antigen may cross-react with related antigens. In certain embodiments, an antibody or antigen-binding domain that binds to an antigen or specifically binds to an antigen does not cross-react with other antigens. An antibody or antigen-binding domain that binds to an antigen or specifically binds to an antigen can be identified, for example, by immunoassay, Octet®, Biacore®, or other techniques known to those of skill in the art. In some embodiments, an antibody or antigen-binding domain binds to an antigen or specifically binds to an antigen if it binds to the antigen with higher affinity than any cross-reactive antigens as determined using experimental techniques such as radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA). Typically, a specific or selective response is at least twice the background signal or noise and may be greater than 10 times the background. For a discussion of binding specificity, see, e.g., Fundamental Immunology 332-36 (Paul, ed., 2d ed. 1989). In certain embodiments, the extent of binding of an antibody or antigen-binding domain to a "non-target" protein is less than about 10% of the binding of the antibody or antigen-binding domain to its specific target antigen, as determined, for example, by fluorescence activated cell sorting (FACS) analysis or RIA. With reference to terms such as "specific binding," "specifically binds to," or "specific for," binding refers to binding that is measurably different from non-specific interactions. Specific binding can be measured, for example, by determining binding of a molecule compared to binding of a control molecule, which is generally a molecule of similar structure that does not have binding activity.For example, specific binding can be determined by competition with a control molecule similar to the target, e.g., an excess of unlabeled target. In this case, specific binding is indicated if binding of the labeled target to the probe is competitively inhibited by the excess of unlabeled target. Antibodies or antigen-binding domains that bind to an antigen include those that can bind to the antigen with sufficient affinity so that the antibody is useful, for example, as a diagnostic or therapeutic agent targeted to the antigen. In certain embodiments, antibodies or antigen-binding domains that bind to an antigen have a dissociation constant (K) of 1000 nM, 800 nM, 500 nM, 250 nM, 100 nM, 50 nM, 10 nM, 5 nM, 4 nM, 3 nM, 2 nM, 1 nM, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, or 0.1 nM or less. D In certain embodiments, the antibody or antigen-binding domain binds to an epitope of an antigen that is conserved between antigens from different species (e.g., between human and cynomolgus monkey species).

[0147] "Binding affinity" generally refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., a binding protein such as an antibody) and its binding partner (e.g., an antigen). Unless otherwise stated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The affinity of a binding molecule X for its binding partner Y is generally determined by the dissociation constant (K D ) Affinity can be measured by common methods known in the art, including those described herein. Low affinity antibodies generally bind antigens slowly and tend to dissociate easily, whereas high affinity antibodies generally bind antigens faster and tend to remain bound longer. Various methods for measuring binding affinity are known in the art, any of which can be used for the purposes of the present disclosure. Specific exemplary embodiments include the following: In one embodiment, "K D " or "K DThe "K value" can be measured by assays known in the art, for example, by binding assays. D can be measured, for example, by RIA using the Fab version of the antibody of interest and its antigen (Chen, et al., J. Mol Biol, 1999, 293:865-81). D or K D The value can be measured by using biolayer interferometry (BLI) or surface plasmon resonance (SPR) assays, for example by Octet® using the Octet® Red 96 system, or by Biacore® using, for example, a Biacore® 2000 or Biacore® 3000. Also referred to as "on rate" or "rate of association" or "association rate" or "k on " can be determined using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, for example, using an Octet® Red 96, Biacore® 2000, or Biacore® 3000 system.

[0148] In certain embodiments, antibodies may comprise "chimeric" sequences in which portions of the heavy and / or light chains are identical to or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of the chains are identical to or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, provided that they exhibit the desired biological activity (U.S. Patent No. 4,816,567, and Morrison, et al., Proc. Natl. Acad. Sci. USA, 1984, 81:6851-55). Chimeric sequences may include humanized sequences.

[0149] In certain embodiments, antibodies may contain portions of "humanized" forms of non-human (e.g., murine) antibodies, which are chimeric antibodies comprising a human immunoglobulin (e.g., recipient antibody) in which native CDR residues are replaced by residues from corresponding CDRs of a non-human species (e.g., donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and capacity. In some cases, one or more FR region residues of the human immunoglobulin are replaced by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues that are not found in the recipient antibody or the donor antibody. These modifications are made to further refine antibody performance. The heavy or light chains of a humanized antibody can comprise substantially all of at least one or more variable regions, in which all or substantially all of the CDRs correspond to those of a non-human immunoglobulin and all or substantially all of the FRs are those of a human immunoglobulin sequence. In certain embodiments, a humanized antibody comprises at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones, et al., Nature, 1986, 321:522-25; Riechmann, et al., Nature, 1988, 332:323-29; Presta, Curr. Op. Struct. Biol., 1992, 2:593-96; Carter, et al., Proc. Natl. Acad. Sci. USA, 1992, 89:4285-89; U.S. Patent Nos. 6,800,738, 6,719,971, 6,639,055, 6,407,213, and 6,054,297.

[0150] In certain embodiments, an antibody can comprise a portion of a "fully human antibody" or "human antibody," which terms are used interchangeably herein and refer to an antibody comprising a human variable region and, for example, a human constant region. In specific embodiments, these terms refer to an antibody comprising variable and constant regions of human origin. A "fully human" antibody can also encompass, in certain embodiments, an antibody that binds a polypeptide and is encoded by a nucleic acid sequence that is a naturally occurring somatic variant of a human germline immunoglobulin nucleic acid sequence. The term "fully human antibody" includes antibodies having variable and constant regions that correspond to human germline immunoglobulin sequences as described by Kabat et al. (See Kabat, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242). A "human antibody" is one that has an amino acid sequence that corresponds to that of an antibody produced by a human and / or has been produced using any of the techniques for producing human antibodies. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using a variety of techniques known in the art, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol., 1991, 227:381; Marks, et al., 1991, J. Mol. Biol., 1991, 222:581) and yeast display libraries (Chao, et al., Nature Protocols, 2006, 1:755-68).Human monoclonal antibodies can also be prepared by the methods described in Cole, et al., Monoclonal Antibodies and Cancer Therapy 77(1985); Boerner, et al., J. Immunol., 1991, 147(1):86-95; and van Dijk and van de Winkel, Curr. Opin. Pharmacol., 2001, 5:368-74. Human antibodies can be prepared by administering antigen to transgenic animals, e.g., mice, that have been modified to produce such antibodies in response to antigen challenge, but whose endogenous gene loci have been disabled (see, e.g., Jakobovits, Curr. Opin. Biotechnol., 1995, 6(5):561-66; Bruggemann and Taussing, Curr. Opin. Biotechnol., 1997, 8(4):455-58; and U.S. Pat. Nos. 6,075,181 and 6,150,584 regarding XENO Mouse™ technology). See also, e.g., Li, et al., Proc. Natl. Acad. Sci. USA, 2006, 103:3557-62 (regarding human antibodies generated via human B cell hybridoma technology).

[0151] In certain embodiments, an antibody can comprise a portion of a "recombinant human antibody," which phrase includes human antibodies prepared, expressed, created, or isolated by recombinant means, e.g., antibodies expressed using a recombinant expression vector transfected into a host cell, antibodies isolated from a recombinant combinatorial human antibody library, antibodies isolated from animals (e.g., mice or cows) that are transgenic and / or transchromosomal for human immunoglobulin genes (e.g., Taylor, LD, et al., Nucl. Acids Res., 1992 20:6287-6295), or antibodies prepared, expressed, created, or isolated by any other means involving splicing human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies can have variable and constant regions derived from human germline immunoglobulin sequences (see Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDapartment of Health and Human Services, NIH Publication No. 91-3242). However, in certain embodiments, such recombinant human antibodies have been subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis), such that the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but are sequences that may not naturally exist within the human antibody germline repertoire in vivo.

[0152] In certain embodiments, an antibody can comprise a portion of a "monoclonal antibody," a term used herein to refer to an antibody obtained from a population of substantially homogeneous antibodies, e.g., the individual antibodies comprising the population are identical except for possible naturally occurring mutations that may be present in minor amounts, and each monoclonal antibody typically recognizes a single epitope on an antigen. In specific embodiments, a "monoclonal antibody," as used herein, is an antibody produced by a single hybridoma or other cell. The term "monoclonal" is not limited to a particular method for making the antibody. For example, monoclonal antibodies useful in the present disclosure can be prepared by the hybridoma method first described by Kohler et al., 1975, Nature 256:495, or can be made using recombinant DNA methods in bacterial or eukaryotic animal or plant cells (see, e.g., U.S. Pat. No. 4,816,567). Monoclonal antibodies can also be isolated from phage antibody libraries using the techniques described, for example, in Clackson, et al., Nature, 1991, 352:624-28 and Marks, et al., J. Mol. Biol., 1991, 222:581-97. Other methods for preparing clonal cell lines and the monoclonal antibodies expressed thereby are well known in the art. See, for example, Short Protocols in Molecular Biology (Ausubel et al. eds., 5th ed. 2002).

[0153] A typical four-chain antibody unit is a heterotetrameric glycoprotein composed of two identical light chains (L) and two identical heavy chains (H). In the case of IgG, the four-chain unit is generally approximately 150,000 daltons. Each L chain is linked to an H chain by one covalent disulfide bond, and the two H chains are linked to each other by one or more disulfide bonds depending on the H chain isotype. Each H and L chain also has regularly spaced intrachain disulfide bridges. Each H chain has a variable domain (VH) at its N-terminus followed by three constant domains (CH) for the α and γ chains, and four CH domains for the μ and ε isotypes. Each L chain has a variable domain (VL) at its N-terminus followed by a constant domain (CL) at its other end. The VL is aligned with the VH, and the CL is aligned with the first constant domain (CH1) of the heavy chain. Particular amino acid residues are believed to form an interface between the light-chain and heavy-chain variable domains. The pairing of VH and VL together forms a single antigen-binding site. For the structure and properties of different classes of antibodies, see, for example, Basic and Clinical Immunology 71 (Stites, et al. eds., 8th ed. 1994); and Immunobiology (Janeway, et al. eds., 5th ed. 2001).

[0154] The term "Fab" or "Fab region" refers to the region of an antibody that binds to an antigen. Conventional IgGs typically contain two Fab regions, each located in one of the two arms of the Y-shaped IgG structure. Each Fab region typically consists of one variable region and one constant region from each of the heavy and light chains. More specifically, the variable and constant regions of the heavy chain in the Fab region are the VH and CH1 regions, and the variable and constant regions of the light chain in the Fab region are the VL and CL regions. The VH, CH1, VL, and CL regions in the Fab region can be arranged in various ways to confer antigen-binding capability according to the present disclosure. For example, the VH and CH1 regions can be on one polypeptide, while the VL and CL regions can be on separate polypeptides, as in the Fab region of a conventional IgG. Alternatively, the VH, CH1, VL, and CL regions can all be present on the same polypeptide and oriented in a different order, as described in more detail in the following sections.

[0155] The terms "variable region," "variable domain," "V region," or "V domain" refer to a portion of an antibody's light or heavy chain, generally located at the amino terminus of the light or heavy chain, approximately 120-130 amino acids in length for heavy chains and approximately 100-110 amino acids in length for light chains, that is used to determine the binding and specificity of each particular antibody to its specific antigen. The variable region of a heavy chain may be referred to as "VH." The variable region of a light chain may be referred to as "VL." The term "variable" refers to the fact that certain segments of the variable region vary significantly in sequence among antibodies. The V region mediates antigen binding and determines the specificity of a particular antibody for a particular antigen. However, the variability is not uniform across the 110-amino acid span of the variable region. Instead, the V region consists of less variable (e.g., relatively invariant) stretches of approximately 15-30 amino acids called framework regions (FRs), separated by shorter regions of greater variability (e.g., extreme variability) called "hypervariable regions," each approximately 9-12 amino acids in length. The heavy and light chain variable regions each contain four FRs, mostly in a β-sheet structure, connected by three hypervariable regions, which form loops connecting the β-sheet structure and, in some cases, form part of the β-sheet structure. The hypervariable regions within each chain are held in close proximity by the FRs and, together with the hypervariable regions of the other chain, contribute to the formation of the antigen-binding site of antibodies (see, e.g., Kabat et al., Sequences of Proteins of Immunological Interest (5th ed. 1991)). The constant regions are not directly involved in binding the antibody to an antigen, but exhibit various effector functions, such as antibody participation in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). The sequences of the variable regions vary significantly among different antibodies. In a specific embodiment, the variable regions are human variable regions.

[0156] The terms "variable region residue numbering according to Kabat" or "amino acid position numbering as in Kabat," and variations thereof, refer to the numbering system used in the heavy or light chain variable regions of the antibody compilations of Kabat et al. (supra). Using this numbering system, the actual linear amino acid sequence may contain fewer or additional amino acids corresponding to shortening of, or insertion into, the FRs or CDRs of the variable domain. For example, a heavy chain variable domain may contain a single amino acid insertion after residue 52 (residue 52a according to Kabat) and three inserted residues after residue 82 (e.g., residues 82a, 82b, and 82c according to Kabat). The Kabat numbering of residues can be determined for a given antibody by aligning the antibody sequence with the "standard" Kabat numbering sequence at the region of homology. The Kabat numbering system is typically used when referring to variable domain residues (approximately residues 1-107 for the light chain and approximately residues 1-113 for the heavy chain) (e.g., Kabat et al., supra). The "EU numbering system" or "EU index" is generally used when referring to residues within an immunoglobulin heavy chain constant region (e.g., the EU index reported in Kabat et al., supra). "EU index as in Kabat" refers to the residue numbering of the human IgG1 EU antibody. Other numbering systems have been described, for example, by AbM, Chothia, Contact, IMGT, and AHon.

[0157] The term "heavy chain" when used with respect to antibodies refers to a polypeptide chain of approximately 50 to 70 kDa, the amino-terminal portion of which contains a variable region of approximately 120 to 130 amino acids, and the carboxy-terminal portion of which contains a constant region. The constant region can be one of five distinct types (e.g., isotypes), designated alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (μ), based on the amino acid sequence of the heavy chain constant region. The distinct heavy chains vary in size, with α, δ, and γ containing approximately 450 amino acids, and μ and ε containing approximately 550 amino acids. When combined with light chains, these distinct types of heavy chains give rise to five well-known classes (e.g., isotypes) of antibodies: IgA, IgD, IgE, IgG, and IgM (including the four subclasses of IgG, i.e., IgG1, IgG2, IgG3, and IgG4).

[0158] The term "light chain" when used in reference to an antibody refers to a polypeptide chain of approximately 25 kDa, which contains a variable region of about 100 to about 110 or more amino acids at its amino terminal end and a constant region at its carboxy terminal end. The approximate length of a light chain is 211 to 217 amino acids. There are two different types, called kappa (κ) or lambda (λ), based on the amino acid sequence of the constant domain.

[0159] As used herein, the terms "hypervariable region," "hypervariable region, HVR," "complementarity-determining region," and "CDR" are used interchangeably. "CDR" refers to one of the three hypervariable regions (H1, H2, or H3) within the non-framework region of an immunoglobulin (Ig or antibody) VH β-sheet framework, or one of the three hypervariable regions (L1, L2, or L3) within the non-framework region of an antibody VL β-sheet framework. Thus, CDRs are variable region sequences interspersed within framework region sequences.

[0160] CDR regions are well known to those skilled in the art and are defined by well-known numbering systems. For example, Kabat complementarity determining regions (CDRs) are based on sequence variability and are the most commonly used (see, e.g., Kabat et al., supra). Chothia, instead, refers to the position of the structural loop (see, e.g., Chothia and Lesk, J. Mol. Biol., 1987, 196:901-17). When numbered using the Kabat numbering rules, the end of the Chothia CDR-H1 loop varies from H32 to H34 depending on the length of the loop (this is because the Kabat numbering scheme places insertions at H35A and H35B; if neither 35A nor 35B is present, the loop ends at 32; if only 35A is present, the loop ends at 33; and if both 35A and 35B are present, the loop ends at 34). AbM hypervariable regions represent a compromise between Kabat CDRs and Chothia structural loops and are used by Oxford Molecular's AbM antibody modeling software (see, e.g., Antibody Engineering Vol. 2 (Kontermann and Dubel, eds., 2nd ed. 2010)). The "contact" hypervariable regions are based on an analysis of available complex crystal structures. Another universal numbering system that has been developed and widely adopted is the ImMunoGeneTics (IMGT) Information System® (Lafranc, et al., Dev. Comp. Immunol., 2003, 27(1):55-77). IMGT is an integrated information system dedicated to immunoglobulins (IGs), T cell receptors (TCRs), and major histocompatibility complexes (MHCs) of humans and other vertebrates. CDRs are referred to herein in terms of both amino acid sequence and location within a light or heavy chain.Because the "location" of CDRs within the structure of immunoglobulin variable domains is conserved among species and resides in structures called loops, CDR and framework residues are readily identified by using a numbering system that aligns variable domain sequences according to structural features. This information can be used to graft and replace CDR residues from one species of immunoglobulin into an acceptor framework, typically from a human antibody. An additional numbering system (AHon) has been developed by Honegger and Pluckthun, J. Mol. Biol., 2001, 309:657-70. For example, the correspondence between numbering systems, including Kabat numbering and the IMGT specific numbering system, is well known to those skilled in the art (see, e.g., Kabat, supra; Chothia and Lesk, supra; Martin, supra; Lefranc, et al., supra). Residues from each of these hypervariable regions or CDRs are shown below.

[0161] [Table 1]

[0162] The boundaries of a given CDR may vary depending on the scheme used for identification. Thus, unless otherwise specified, the terms "CDR" and "complementarity determining region" of a given antibody or region thereof, such as a variable region, and individual CDRs of an antibody or region thereof (e.g., "CDR-H1, CDR-H2"), should be understood to encompass the complementarity determining regions defined by any of the known schemes described hereinabove. In some cases, a scheme for identifying a particular CDR or CDRs is specified, such as CDRs defined by the Kabat, Chothia, or Contact methods. In other cases, the specific amino acid sequences of the CDRs are provided.

[0163] The hypervariable regions may include "extended hypervariable regions" such as: 24-36 or 24-34 (L1), 46-56 or 50-56 (L2), and 89-97 or 89-96 (L3) in VL, and 26-35 or 26-35A (H1), 50-65 or 49-65 (H2), and 93-102, 94-102, or 95-102 (H3) in VH.

[0164] The term "constant region" or "constant domain" refers to the carboxy-terminal portions of the light and heavy chains that are not directly involved in binding the antibody to an antigen, but which exhibit various effector functions, such as interaction with Fc receptors. This term refers to the portion of an immunoglobulin molecule that has a more conserved amino acid sequence compared to the other portion of the immunoglobulin, the variable region, which contains the antigen-binding site. The constant region can include the CH1, CH2, and CH3 regions of the heavy chain and the CL region of the light chain.

[0165] The term "framework" or "FR" refers to the variable region residues adjacent to the CDRs. FR residues are present, for example, in chimeric antibodies, humanized antibodies, human antibodies, domain antibodies, diabodies, linear antibodies, and bispecific antibodies. FR residues are variable domain residues other than hypervariable region or CDR residues. Typically, there are four FR regions in each of the VH and VL regions. The FR regions in VH are VH FR1, VH FR2, VH FR3, and VH FR4 (or FR H1, FR H2, FR H3, and FR H4). The FR regions in VL are VL FR1, VL FR2, VL FR3, and VL FR4 (or FR L1, FR L2, FR L3, and FR L4).

[0166] As used herein, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including, for example, native-sequence Fc regions, recombinant Fc regions, and variant Fc regions. While the boundaries of the Fc region of an immunoglobulin heavy chain can vary, the Fc region of a human IgG heavy chain is often defined as stretching from the amino acid residue at position Cys226, or from the amino acid residue at position Pro230, to the carboxyl terminus. The C-terminal lysine of the Fc region (residue 447 according to the EU numbering system) can be removed, for example, during antibody production or purification, or by recombinantly engineering the nucleic acid encoding the antibody heavy chain. Thus, an intact antibody composition can include an antibody population in which all K447 residues have been removed, an antibody population in which the K447 residue has not been removed, and an antibody population having a mixture of antibodies with and without the K447 residue. A "functional Fc region" possesses the "effector functions" of a native-sequence Fc region. Exemplary "effector functions" include C1q binding, CDC, Fc receptor binding, ADCC, phagocytosis, down-regulation of cell surface receptors (e.g., B cell receptors), and the like. Such effector functions generally require that the Fc region be combined with a binding region or domain (e.g., an antibody variable region or domain) and can be assessed using a variety of assays known to those of skill in the art. A "variant Fc region" comprises an amino acid sequence that differs from that of a native-sequence Fc region by at least one amino acid modification (e.g., substitution, addition, or deletion). In certain embodiments, the variant Fc region comprises at least one amino acid substitution compared to a native-sequence Fc region or the Fc region of a parent polypeptide, e.g., from about 1 to about 10 amino acid substitutions, or from about 1 to about 5 amino acid substitutions in the native-sequence Fc region or the Fc region of a parent polypeptide. The variant Fc region herein may have at least about 80% homology to a native sequence Fc region and / or the Fc region of a parent polypeptide, or at least about 90% homology thereto, for example, at least about 95% homology thereto.

[0167] The term "variant," when used with respect to an antigen or antibody, can refer to a peptide or polypeptide that contains one or more (e.g., about 1 to about 25, about 1 to about 20, about 1 to about 15, about 1 to about 10, or about 1 to about 5) amino acid sequence substitutions, deletions, and / or additions compared to the native or unmodified sequence.

[0168] The term "identity" refers to the relationship between the sequences of two or more polypeptide molecules or two or more nucleic acid molecules, as determined by aligning and comparing the sequences. "Percent (%) amino acid sequence identity" to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical to those in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, without considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved by a variety of methods within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN (DNAStar, Inc.) software. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms necessary to achieve maximum alignment over the full length of the sequences being compared.

[0169] A "modification" of an amino acid residue / position refers to a change in the primary amino acid sequence compared to the starting amino acid sequence, where the change results from a sequence change involving that amino acid residue / position. For example, typical modifications include substitution of the residue with another amino acid (e.g., a conservative or non-conservative substitution), insertion of one or more (e.g., generally less than 5, 4, or 3) amino acids adjacent to that residue / position, and / or deletion of that residue / position.

[0170] As used herein, "epitope" is a term of art that refers to a localized region of an antigen to which an antibody can specifically bind. An epitope can be a linear or conformational epitope, a non-linear epitope, or a discontinuous epitope. In the case of a polypeptide antigen, for example, an epitope can be consecutive amino acids of a polypeptide (a "linear" epitope), or it can include amino acids from two or more non-contiguous regions of a polypeptide (a "conformational," "non-linear," or "discontinuous" epitope). In general, it will be understood by those skilled in the art that a linear epitope may or may not depend on secondary, tertiary, or quaternary structure. For example, in some embodiments, an antibody binds to a group of amino acids regardless of whether the amino acids are folded into the native three-dimensional protein structure. In other embodiments, antibodies require that the amino acid residues comprising the epitope exhibit a particular conformation (eg, a bend, twist, turn, or fold) in order to recognize and bind to the epitope.

[0171] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, may comprise modified amino acids, and may be interrupted by non-amino acids. The term also refers to amino acid polymers that are naturally modified or modified by intervention, such as disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification. The definition also includes polypeptides containing one or more analogs of an amino acid, including, but not limited to, unnatural amino acids, as well as other modifications known in the art. Because the polypeptides of the present disclosure may be based on antibodies or other members of the immunoglobulin superfamily, it is understood that in certain embodiments, a "polypeptide" can occur as a single chain or as two or more related chains.

[0172] The term "vector" refers to a substance used to carry or contain a nucleic acid sequence, such as a nucleic acid sequence encoding an antibody described herein, for introducing the nucleic acid sequence into a host cell. Vectors applicable for use include, for example, expression vectors, plasmids, phage vectors, viral vectors, episomes, and artificial chromosomes, which may contain selection sequences or markers capable of stable integration into a host cell chromosome. In addition, a vector may contain one or more selectable marker genes and appropriate expression control sequences. Selectable marker genes that may be included provide, for example, resistance to antibiotics or toxins, complement deficiencies in auxiliary auxotrophies, or supply critical nutrients absent from the culture medium. Expression control sequences may include constitutive and inducible promoters, transcription enhancers, transcription terminators, and the like, as are well known in the art. When two or more nucleic acid molecules are coexpressed (e.g., both the heavy and light chains of an antibody, or antibody VH and VL), both nucleic acid molecules may be inserted, for example, into a single expression vector or into separate expression vectors. For expression in a single vector, the encoding nucleic acids can be operably linked to a common expression control sequence or to different expression control sequences, such as one inducible promoter and one constitutive promoter. Introduction of nucleic acid molecules into host cells can be confirmed using methods well known in the art. Such methods include, for example, nucleic acid analysis, such as Northern blot or amplification of mRNA by polymerase chain reaction (PCR), immunoblotting for expression of gene products, or other suitable analytical methods for testing expression of the introduced nucleic acid sequence or its corresponding gene product. Those skilled in the art will understand that nucleic acid molecules will be expressed in amounts sufficient to produce the desired product, and will further understand that expression levels can be optimized to obtain sufficient expression using methods well known in the art.

[0173] As used herein, the term "host" refers to an animal, such as a mammal (e.g., a human).

[0174] As used herein, the term "host cell" refers to a particular subject cell that can be transfected with a nucleic acid molecule, and the progeny or potential progeny of such a cell. The progeny of such a cell may not be identical to the parent cell transfected with the nucleic acid molecule due to mutations or environmental influences that may occur during the subsequent generation or integration of the nucleic acid molecule into the host cell genome.

[0175] An "isolated nucleic acid" is a nucleic acid, e.g., RNA, DNA, or mixed nucleic acid, that is substantially separated from other genomic DNA sequences and proteins or complexes, such as ribosomes and polymerases, that are naturally associated with the native sequence. An "isolated" nucleic acid molecule is one that is separated from other nucleic acid molecules that are present in the nucleic acid molecule's natural source. Furthermore, an "isolated" nucleic acid molecule, such as a cDNA molecule, may be substantially free of other cellular material or culture medium if produced by recombinant techniques, or substantially free of chemical precursors or other chemicals if chemically synthesized. In specific embodiments, one or more nucleic acid molecules encoding an antibody described herein are isolated or purified. This term encompasses nucleic acid sequences that have been removed from their naturally occurring environment, and includes recombinant or cloned DNA isolates, as well as chemically synthesized analogs or heterologous biologically synthesized analogs. A substantially pure molecule can include isolated forms of the molecule.

[0176] As used interchangeably herein, "polynucleotide," "nucleotide," or "nucleic acid" refers to a polymer of nucleotides of any length, including DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase or by a synthetic reaction. Polynucleotides can include modified nucleotides, such as methylated nucleotides and their analogs. As used herein, "oligonucleotide" refers to a short, generally single-stranded, synthetic polynucleotide, generally, but not necessarily, less than about 200 nucleotides in length. The terms "oligonucleotide" and "polynucleotide" are not mutually exclusive. The above description of polynucleotides is equally fully applicable to oligonucleotides. Cells producing antibodies of the present disclosure can include parent hybridoma cells, as well as bacterial and eukaryotic host cells into which nucleic acid encoding the antibody has been introduced. Unless otherwise specified, the left-hand end of any single-stranded polynucleotide sequence disclosed herein is the 5' end. The left-hand direction of a double-stranded polynucleotide sequence is referred to as the 5' direction. The direction of 5' to 3' addition of nascent RNA transcripts is referred to as the transcription direction. The region of the sequence on the DNA strand that has the same sequence as the RNA transcript at the 5' to 5' end of the RNA transcript is referred to as the "upstream sequence," and the region of the sequence on the DNA strand that has the same sequence as the RNA transcript at the 3' to 3' end of the RNA transcript is referred to as the "downstream sequence."

[0177] As used herein, the term "multispecific antibody" refers to an antibody comprising a plurality of immunoglobulin variable domain sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In certain embodiments, the first and second epitopes do not overlap or substantially do not overlap. In certain embodiments, the first and second epitopes are on different antigens, e.g., on different proteins (or different subunits of a multimeric protein). In certain embodiments, the multispecific antibody comprises a third, fourth, or fifth immunoglobulin variable domain. In certain embodiments, the multispecific antibody is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.

[0178] As used herein, the term "bispecific antibody" refers to a multispecific antibody that binds to no more than two epitopes or two antigens. A bispecific antibody is characterized by a first immunoglobulin variable domain sequence that has binding specificity for a first epitope (e.g., an epitope on an NKG2d or NKp46 antigen) and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope (e.g., a tumor-associated antigen (e.g., a BCMA or GPRC5d antigen). In some embodiments, the first and second epitopes are on different antigens, e.g., different proteins (or different subunits of a multimeric protein). In some embodiments, a bispecific antibody comprises heavy chain and light chain variable domain sequences that have binding specificity for a first epitope and heavy chain and light chain variable domain sequences that have binding specificity for a second epitope. In some embodiments, a bispecific antibody comprises heavy chain and light chain variable domain sequences that have binding specificity for a first epitope and heavy chain and light chain variable domain sequences that have binding specificity for a second epitope. A heterospecific antibody comprises a half antibody or fragment thereof having binding specificity for a first epitope and a half antibody or fragment thereof having binding specificity for a second epitope. In some embodiments, a bispecific antibody comprises an scFv or fragment thereof having binding specificity for a first epitope and an scFv or fragment thereof having binding specificity for a second epitope. In some embodiments, the first epitope is located on NKG2d and the second epitope is located on BCMA. In some embodiments, the first epitope is located on NKG2d and the second epitope is located on GPRC5d. In some embodiments, the first epitope is located on NKp46 and the second epitope is located on BCMA. In some embodiments, the first epitope is located on NKp46 and the second epitope is located on GPRC5d.

[0179] As used herein, the term "NKG2d" refers to the NKG2-D type II integral membrane protein. NKG2d may also be referred to as "NK cell receptor D," "NKG2-D activating NK receptor," or "CD314." The term "NKG2d," unless otherwise specified, includes any NKG2d variant, isoform, and species homolog that is naturally expressed by cells (including NK cells) or that can be expressed on cells transfected with a gene or cDNA encoding the polypeptide; in certain embodiments, "NKG2d" is human NKG2d.

[0180] As used herein, the term "NKp46" refers to natural killer cell p46-related protein. NKp46 may also be referred to as "natural cytotoxicity-triggering receptor 1," "NK-p46," or "CD335." The term "NKp46," unless otherwise specified, includes any NKp46 variant, isoform, and species homolog that is naturally expressed by cells (including NK cells) or that can be expressed on cells transfected with a gene or cDNA encoding the polypeptide; in certain embodiments, "NKp46" is human NKp46.

[0181] As used herein, the term "BCMA" refers to B-cell maturation antigen, also known as TNFRSF17 or CD269, which is a member of the tumor necrosis factor receptor (TNFR) superfamily. Unless otherwise specified, the term "BCMA" includes any BCMA variant, isoform, and species homolog that is naturally expressed by a cell (including a cancer cell) or that can be expressed on a cell transfected with a gene or cDNA encoding the polypeptide; in certain embodiments, "BCMA" is human BCMA.

[0182] As used herein, the term "GPRC5d" refers to G protein-coupled receptor family C group 5 member D. Unless otherwise specified, the term "GPRC5d" includes any GPRC5d variants, isoforms, and species homologs that are naturally expressed by cells (including cancer cells) or that can be expressed on cells transfected with a gene or cDNA encoding the polypeptide; in certain embodiments, "GPRC5d" is human GPRC5d.

[0183] As used herein, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency for use in animals, and more particularly for use in humans, or listed in the United States Pharmacopoeia, the European Pharmacopoeia, or other generally recognized pharmacopoeias.

[0184] The term "excipient" refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, solvent, encapsulating material, etc. Excipients include, for example, encapsulating materials or additives such as absorption enhancers, antioxidants, binders, buffers, carriers, coating agents, colorants, diluents, disintegrants, emulsifiers, expanders, fillers, flavoring agents, wetting agents, lubricants, fragrances, preservatives, propellants, release agents, bactericides, sweeteners, solubilizers, wetting agents, and mixtures thereof. The term "excipient" can also refer to a diluent, adjuvant (e.g., Freund's adjuvant (complete or incomplete)), or vehicle.

[0185] In some embodiments, the excipient is a pharmaceutically acceptable excipient. Examples of pharmaceutically acceptable excipients include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (e.g., less than about 10 amino acid residues) polypeptides; proteins such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrin; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or non-ionic surfactants such as TWEEN™, polyethylene glycol (PEG), and PLURONICS™. Other examples of pharmaceutically acceptable excipients are described in Remington and Gennaro, Remington's Pharmaceutical Sciences (18th ed. 1990).

[0186] In one embodiment, each component is "pharmaceutically acceptable" in the sense of being compatible with the other components of the pharmaceutical formulation and suitable for use in contact with the tissues or organs of humans and animals without undue toxicity, irritation, allergic response, immunogenicity, or other problem or complication, commensurate with a reasonable benefit / risk ratio. See, e.g., Lippincott Williams & Wilkins: Philadelphia, PA, 2005; Handbook of Pharmaceutical Excipients, 6th ed.; Rowe et al., Eds.; The Pharmaceutical Press and the American Pharmaceutical Association: 2009; Handbook of Pharmaceutical Additives, 3rd ed.; Ash and Ash Eds.; Gower Publishing Company: 2007; Pharmaceutical Preformulation and Formulation, 2nd ed.; Gibson Ed.; CRC Press LLC: Boca Raton, FL, 2009. In some embodiments, a pharmaceutically acceptable excipient is nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. In some embodiments, the pharmaceutically acceptable excipient is an aqueous pH buffered solution.

[0187] In some embodiments, the excipient may be a sterile liquid, such as water or oil, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, or sesame oil. Water is an exemplary excipient when the composition (e.g., a pharmaceutical composition) is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions may also be employed as liquid excipients, particularly for injectable solutions. Excipients may also include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, dried skim milk, glycerin, propylene, glycol, water, ethanol, or the like. If desired, the composition may further contain minor amounts of wetting or emulsifying agents, or pH buffering agents. The composition may take the form of a solution, suspension, emulsion, tablet, pill, capsule, powder, sustained-release formulation, or the like. Oral composition containing formulations can include standard excipients such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, etc.

[0188] A composition containing a pharmaceutical compound can include, for example, an antibody in isolated or purified form, together with a suitable amount of excipients.

[0189] As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount of an antibody or pharmaceutical composition provided herein that is sufficient to bring about a desired result.

[0190] The terms "subject" and "patient" can be used interchangeably herein. As used herein, in certain embodiments, a subject is a mammal, such as a non-primate (e.g., cows, pigs, horses, cats, dogs, rats, etc.) or a primate (e.g., monkeys and humans). In specific embodiments, the subject is a human. In one embodiment, the subject is a mammal, e.g., a human, diagnosed with a condition or disorder. In another embodiment, the subject is a mammal, e.g., a human, at risk of developing a condition or disorder.

[0191] "Administering" or "administration" refers to the act of injecting or otherwise physically delivering a substance present outside the body to a patient, such as by mucosal, intradermal, intravenous, intramuscular, subcutaneous delivery, and / or any other physical delivery method described herein or known in the art.

[0192] As used herein, the terms "treat / treat," "treatment," and "treating" refer to the reduction or amelioration of the progression, severity, and / or duration of a disease or condition resulting from the administration of one or more therapies. Treatment may be determined by assessing whether there has been a reduction, alleviation, and / or alleviation of one or more symptoms associated with the underlying disease, such that an improvement is observed in the patient, even though the patient may still be suffering from the underlying disease. The term "treat / treating" includes both management and remission of disease. The terms "manage," "managing," and "management" refer to the beneficial effects a subject derives from a treatment, which does not necessarily result in a cure of the disease.

[0193] The terms "prevent," "preventing," and "prevention" refer to reducing the likelihood of the onset (or recurrence) of a disease, disorder, condition, or associated symptom.

[0194] The terms "about" and "approximately" refer to within 20%, within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1%, or less of a given value or range.

[0195] As used in this disclosure and the claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0196] Whenever an embodiment is described herein with the term "comprising," it is understood that other similar embodiments otherwise described in terms of "consisting of" and / or "consisting essentially of" are also provided. Whenever an embodiment is described herein with the phrase "consisting essentially of," it is also understood that similar embodiments otherwise described in terms of "consisting of" are also provided.

[0197] The term "between" used in phrases such as "between A and B" or "between A and B" refers to a range that includes both A and B.

[0198] As used herein, the term "and / or" in phrases such as "A and / or B" is intended to include both A and B; A or B; A alone; and B alone. Similarly, the term "and / or" in phrases such as "A, B, and / or C" is intended to include each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A alone; B alone; and C alone.

[0199] binding molecule NK cell binding molecule In one aspect, provided herein are binding molecules, such as antibodies or fragments thereof, that bind to a cell surface antigen of an NK cell. In some embodiments, the antigen is NKG2d. In some embodiments, the antigen is NKp46.

[0200] In one aspect, provided herein is an antibody that binds to NKG2d. In some embodiments, the antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKG2d antibody is not a single domain antibody or a nanobody. In some embodiments, the NKG2d antibody is a humanized antibody.

[0201] In some embodiments, provided herein are antibodies that specifically bind to NKG2d and can regulate NK cell activity. In some embodiments, the NKG2d antibodies provided herein can regulate NK cell assembly. In some embodiments, the NKG2d antibodies provided herein can activate NK cells. In certain embodiments, the NK cells are human NK cells.

[0202] In certain embodiments, provided herein are anti-NKG2d antibodies comprising the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d antibodies comprising the VH region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d antibodies comprising the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d antibodies comprising the VH region of any one of the antibodies described herein and the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d antibodies comprising the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d antibodies comprising the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d antibodies comprising the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein and the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein. Representative VH and VL amino acid sequences, including the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 amino acid sequences of the NKG2d antibodies provided herein, are provided in the Sequence Listing and Tables 3, 4, 7, and 8.

[0203] In some embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody is an IgG antibody. In other embodiments, the antibody is an IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the antibody is a bispecific antibody. In certain embodiments, the antibody is multivalent. In other embodiments, the antibody is capable of binding to at least three antigens. In some embodiments, the antibody is capable of binding to at least five antigens.

[0204] In certain embodiments, an anti-NKG2d antibody is provided that is an intact antibody. In other embodiments, an anti-NKG2d antibody that is an antigen-binding fragment of an anti-NKG2d antibody is provided. In some embodiments, the antigen-binding fragment of an anti-NKG2d antibody is a functional fragment. In some embodiments, the antigen-binding fragment is a diabody. In some embodiments, the antigen-binding fragment is a Fab. In some embodiments, the antigen-binding fragment is a Fab'. In some embodiments, the antigen-binding fragment is a F(ab')2. In some embodiments, the antigen-binding fragment is an Fv fragment. In some embodiments, the antigen-binding fragment is a disulfide-stabilized Fv fragment (dsFv). In some embodiments, the antigen-binding fragment is a (dsFv)2. In some embodiments, the antigen-binding fragment is a bispecific dsFv (dsFv-dsFv'). In some embodiments, the antigen-binding fragment is a disulfide-stabilized diabody (dsdiabody). In some embodiments, the antigen-binding fragment is a single-chain antibody molecule (scFv). In some embodiments, the antigen-binding fragment is a single domain antibody (sdAb). In some embodiments, the antigen-binding fragment is an scFv dimer (bivalent diabody). In some embodiments, the antigen-binding fragment is a multispecific antibody formed from a portion of an antibody comprising one or more CDRs. In some embodiments, the antigen-binding fragment is a domain antibody. In some embodiments, the antigen-binding fragment is a bivalent domain antibody. In some embodiments, the antigen-binding fragment is an antibody fragment that binds to an antigen but does not comprise a complete antibody structure.

[0205] In certain embodiments, the NKG2d antibody comprises a VH region and a VL region. In some embodiments, the NKG2d antibody is a single-chain antibody. In some embodiments, the NKG2d antibody is a single-domain antibody. In some embodiments, the antigen-binding fragment is a camelized single-domain antibody. In some embodiments, the NKG2d antibody is a nanobody. In certain embodiments, the NKG2d antibody is a VHH antibody. In certain embodiments, the NKG2d antibody is a llama antibody. In some embodiments, the NKG2d antibody is not a single-chain antibody. In some embodiments, the NKG2d antibody is not a single-domain antibody. In some embodiments, the NKG2d antibody is not a nanobody. In certain embodiments, the NKG2d antibody is not a VHH antibody. In certain embodiments, the NKG2d antibody is not a llama antibody. In some embodiments, the NKG2d antibody is a multispecific antibody. In other embodiments, the NKG2d is a bispecific antibody. In certain embodiments, the multispecific antibody comprises an antigen-binding fragment of an NKG2d antibody provided herein. In other embodiments, the bispecific antibody comprises an antigen-binding fragment of an NKG2d antibody provided herein. In some embodiments, the NKG2d antibody is an agonist antibody. In certain embodiments, the NKG2d antibody activates NK cells. In some embodiments, the NKG2d antibody modulates the activity of NK cells. In some embodiments, the NKG2d antibody neither activates nor inactivates the activity of NK cells. In certain embodiments, the NK cells are human NK cells.

[0206] In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to the Kabat numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to the Chothia numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to an exemplary numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to the Contact numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are according to the IMGT numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are according to the AbM numbering system. Exemplary sets of six CDRs (VH CDR1-3 and VL CDR1-3) for certain antibody embodiments are provided herein. Other CDR sets are contemplated and are within the scope of the antibody embodiments provided herein.

[0207] In some embodiments, the antibodies provided herein bind to NKG2d. In some embodiments, antibodies that bind to NKG2d comprise a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 2. In some embodiments, antibodies that bind to NKG2d comprise a VL that includes a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 3. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 2, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 3. In some embodiments, CDR1, CDR2, or CDR3 is determined according to the Kabat numbering scheme, the IMGT numbering scheme, the AbM numbering scheme, the Chothia numbering scheme, the Contact numbering scheme, or a combination thereof. In some embodiments, an antibody that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 2. In some embodiments, an antibody that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments, an antibody that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO:2 and a VL having the amino acid sequence of SEQ ID NO:3.

[0208] In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 13, 14, and 15, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 16, 17, and 18, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 19, 20, and 21, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 22, 23, and 24, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 25, 26, and 27, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 28, 29, and 30, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 31, 32, and 33, respectively.

[0209] In some embodiments, the antibody further comprises one or more framework regions of SEQ ID NO: 2 and / or SEQ ID NO: 3. In some embodiments, the antibodies provided herein are humanized antibodies. The framework regions described herein are determined based on the boundaries of the CDR numbering system. In other words, when CDRs are determined by, for example, Kabat, IMGT, or Chothia, the framework regions are the amino acid residues surrounding the CDRs of the variable region from N-terminus to C-terminus, i.e., in the format FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. For example, FR1 is defined as the amino acid residues N-terminal to the CDR1 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR2 is defined as the amino acid residues between the CDR1 and CDR2 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR3 is defined as the amino acid residues between the CDR2 and CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; and FR4 is defined as the amino acid residues C-terminal to the CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system.

[0210] In some embodiments, antibodies that bind to NKG2d comprise a VH that includes a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 34. In some embodiments, antibodies that bind to NKG2d comprise a VL that includes a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 35. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 34, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 35. In some embodiments, CDR1, CDR2, or CDR3 is determined according to the Kabat numbering scheme, the IMGT numbering scheme, the AbM numbering scheme, the Chothia numbering scheme, the Contact numbering scheme, or a combination thereof. In some embodiments, an antibody that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34. In some embodiments, an antibody that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments, an antibody that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO:34 and a VL having the amino acid sequence of SEQ ID NO:35.

[0211] In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 36, 37, and 38, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 39, 40, and 41, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 42, 43, and 44, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 45, 46, and 47, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 48, 49, and 50, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 51, 52, and 53, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 54, 55, and 56, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 57, 58, and 59, respectively. In some embodiments, an antibody that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 60, 61, and 62, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 63, 64, and 65, respectively.

[0212] In some embodiments, the antibody further comprises one or more framework regions of SEQ ID NO: 34 and / or SEQ ID NO: 35. In some embodiments, the antibodies provided herein are humanized antibodies. The framework regions described herein are determined based on the boundaries of the CDR numbering system. In other words, when the CDRs are determined by, for example, Kabat, IMGT, or Chothia, the framework regions are the amino acid residues surrounding the CDRs of the variable region from N-terminus to C-terminus, i.e., in the format FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. For example, FR1 is defined as the amino acid residues N-terminal to the CDR1 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR2 is defined as the amino acid residues between the CDR1 and CDR2 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR3 is defined as the amino acid residues between the CDR2 and CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; and FR4 is defined as the amino acid residues C-terminal to the CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system.

[0213] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein comprise an amino acid sequence that has a certain percentage of identity compared to any one of the above antibodies.

[0214] The determination of percent identity between two sequences (e.g., amino acid sequences or nucleic acid sequences) can be accomplished using a mathematical algorithm. A non-limiting example of a mathematical algorithm utilized for comparing two sequences is the algorithm of Karlin and Altschul, Proc. Natl. Acad. Sci. USA 87:2264-2268 (1990), modified as in Karlin and Altschul, Proc. Natl. Acad. Sci. USA 90:5873-5877 (1993). Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul et al., J. Mol. Biol. 215:403 (1990). BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, for example, to score=100 and word length=12 to obtain nucleotide sequences homologous to the nucleic acid molecules described herein. BLAST protein searches can be performed with the XBLAST program parameters set, for example, to score 50 and word length = 3, to obtain amino acid sequences homologous to the protein molecules described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be used as described in Altschul et al., Nucleic Acids Res. 25:3389-3402 (1997). Alternatively, PSI BLAST can be used to perform an iterated search that detects distant relationships between molecules (ibid.). When using BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used (see, e.g., the National Center for Biotechnology Information (NCBI) on the World Wide Web at ncbi.nlm.nih.gov). Another non-limiting example of a mathematical algorithm used for sequence comparison is the algorithm of Myers and Miller, CABIOS 4:11-17 (1998).Such an algorithm is incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package. When utilizing the ALIGN program to compare amino acid sequences, a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4 can be used. The percent identity between two sequences can be determined using techniques similar to those described above, with or without allowing gaps. Typically, only exact matches are counted when calculating percent identity.

[0215] In some embodiments, anti-NKG2d antibodies are provided that comprise a VH having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 2. In some embodiments, anti-NKG2d antibodies are provided that comprise a VL having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 3. In some embodiments, an anti-NKG2d antibody is provided that includes a VH having at least about any one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:2, and a VL having at least about any one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO:3.

[0216] In some embodiments, anti-NKG2d antibodies are provided that comprise a VH having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 34. In some embodiments, anti-NKG2d antibodies are provided that comprise a VL having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 35. In some embodiments, an anti-NKG2d antibody is provided that includes a VH having at least about any one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 34, and a VL having at least about any one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 35.

[0217] In some embodiments, a VH or VL sequence having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a reference sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but antibodies containing the sequence retain the ability to bind to NKG2d. In some embodiments, a total of 1 to 10 amino acids are substituted, inserted, and / or deleted. In some embodiments, the substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., within the FRs).

[0218] In another aspect, provided herein is an antibody that competes with any of the NKG2d antibodies described herein for binding to NKG2d. In another aspect, provided herein is an antibody that binds to the same epitope as any of the NKG2d antibodies described herein. In another aspect, provided is an NKG2d antibody that binds to an epitope on NKG2d that overlaps with the epitope on NKG2d bound by the NKG2d antibodies described herein.

[0219] In one embodiment, an antibody is provided that competes with an NKG2d reference antibody for binding to NKG2d. In another embodiment, an NKG2d antibody is provided that binds to the same NKG2d epitope as the NKG2d reference antibody. In another embodiment, an NKG2d antibody is provided that binds to an epitope on NKG2d that overlaps with the epitope on NKG2d bound by the NKG2d reference antibody.

[0220] In one embodiment, the NKG2d reference antibody comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 2, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 3. In one embodiment, the NKG2d reference antibody comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 34, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 35.

[0221] In one aspect, provided herein is an antibody that binds to NKp46. In some embodiments, the antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKp46 antibody is not a single domain antibody or a nanobody. In some embodiments, the NKp46 antibody is a humanized antibody.

[0222] In some embodiments, provided herein are antibodies that specifically bind to NKp46 and can modulate NK cell activity. In some embodiments, the NKp46 antibodies provided herein can modulate NK cell assembly. In some embodiments, the NKp46 antibodies provided herein can activate NK cells. In certain embodiments, the NK cells are human NK cells.

[0223] In certain embodiments, provided herein are anti-NKp46 antibodies comprising the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 antibodies comprising the VH region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 antibodies comprising the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 antibodies comprising the VH region of any one of the antibodies described herein and the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 antibodies comprising the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 antibodies comprising the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 antibodies comprising the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein and the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein. Representative VH and VL amino acid sequences, including the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 amino acid sequences of the NKp46 antibodies provided herein, are provided in the Sequence Listing and Tables 14-15.

[0224] In some embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody is an IgG antibody. In other embodiments, the antibody is an IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the antibody is a bispecific antibody. In certain embodiments, the antibody is multivalent. In other embodiments, the antibody is capable of binding to at least three antigens. In some embodiments, the antibody is capable of binding to at least five antigens.

[0225] In certain embodiments, an NKp46 antibody is provided that is an intact antibody. In other embodiments, an NKp46 antibody is provided that is an antigen-binding fragment of the NKp46 antibody. In some embodiments, the antigen-binding fragment of the NKp46 antibody is a functional fragment. In some embodiments, the antigen-binding fragment is a diabody. In some embodiments, the antigen-binding fragment is a Fab. In some embodiments, the antigen-binding fragment is a Fab'. In some embodiments, the antigen-binding fragment is a F(ab')2. In some embodiments, the antigen-binding fragment is an Fv fragment. In some embodiments, the antigen-binding fragment is a disulfide-stabilized Fv fragment (dsFv). In some embodiments, the antigen-binding fragment is a (dsFv)2. In some embodiments, the antigen-binding fragment is a bispecific dsFv (dsFv-dsFv'). In some embodiments, the antigen-binding fragment is a disulfide-stabilized diabody (dsdiabody). In some embodiments, the antigen-binding fragment is a single-chain antibody molecule (scFv). In some embodiments, the antigen-binding fragment is a single domain antibody (sdAb). In some embodiments, the antigen-binding fragment is an scFv dimer (bivalent diabody). In some embodiments, the antigen-binding fragment is a multispecific antibody formed from a portion of an antibody comprising one or more CDRs. In some embodiments, the antigen-binding fragment is a domain antibody. In some embodiments, the antigen-binding fragment is a bivalent domain antibody. In some embodiments, the antigen-binding fragment is an antibody fragment that binds to an antigen but does not comprise a complete antibody structure.

[0226] In certain embodiments, the NKp46 antibody comprises a VH region and a VL region. In some embodiments, the NKp46 antibody is a single-chain antibody. In some embodiments, the NKp46 antibody is a single-domain antibody. In some embodiments, the antigen-binding fragment is a camelized single-domain antibody. In some embodiments, the NKp46 antibody is a nanobody. In certain embodiments, the NKp46 antibody is a VHH antibody. In certain embodiments, the NKp46 antibody is a llama antibody. In some embodiments, the NKp46 antibody is not a single-chain antibody. In some embodiments, the NKp46 antibody is not a single-domain antibody. In some embodiments, the NKp46 antibody is not a nanobody. In certain embodiments, the NKp46 antibody is not a VHH antibody. In certain embodiments, the NKp46 antibody is not a llama antibody. In some embodiments, the NKp46 antibody is a multispecific antibody. In other embodiments, the NKp46 is a bispecific antibody. In certain embodiments, the multispecific antibody comprises an antigen-binding fragment of an NKp46 antibody provided herein. In other embodiments, the bispecific antibody comprises an antigen-binding fragment of an NKp46 antibody provided herein. In some embodiments, the NKp46 antibody is an agonist antibody. In certain embodiments, the NKp46 antibody activates NK cells. In some embodiments, the NKp46 antibody modulates the activity of NK cells. In some embodiments, the NKp46 antibody neither activates nor inactivates the activity of NK cells. In certain embodiments, the NK cells are human NK cells.

[0227] In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to the Kabat numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to the Chothia numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to an exemplary numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are numbered according to the Contact numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are according to the IMGT numbering system. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 sequences are according to the AbM numbering system. Exemplary sets of six CDRs (VH CDR1-3 and VL CDR1-3) for certain antibody embodiments are provided herein. Other CDR sets are contemplated and are within the scope of the antibody embodiments provided herein.

[0228] In some embodiments, the antibodies provided herein bind to NKp46. In some embodiments, the antibodies that bind to NKp46 comprise a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 67. In some embodiments, the antibodies that bind to NKp46 comprise a VL that includes a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 68. In some embodiments, an antibody that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 67, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 68. In some embodiments, CDR1, CDR2, or CDR3 is determined according to the Kabat numbering scheme, the IMGT numbering scheme, the AbM numbering scheme, the Chothia numbering scheme, the Contact numbering scheme, or a combination thereof. In some embodiments, an antibody that binds to NKp46 comprises a VH having the amino acid sequence of SEQ ID NO: 67. In some embodiments, an antibody that binds to NKp46 comprises a VL having the amino acid sequence of SEQ ID NO: 68. In some embodiments, an antibody that binds NKp46 comprises a VH having the amino acid sequence of SEQ ID NO:67 and a VL having the amino acid sequence of SEQ ID NO:68.

[0229] In some embodiments, an antibody that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 69, 70, and 71, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 72, 73, and 74, respectively. In some embodiments, an antibody that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 75, 76, and 77, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 78, 79, and 80, respectively. In some embodiments, an antibody that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 81, 82, and 83, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 84, 85, and 86, respectively. In some embodiments, an antibody that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 87, 88, and 89, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively. In some embodiments, an antibody that binds to NKp46 comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 93, 94, and 95, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 96, 97, and 98, respectively.

[0230] In some embodiments, the antibody further comprises one or more framework regions of SEQ ID NO: 67 and / or SEQ ID NO: 68. In some embodiments, the antibodies provided herein are humanized antibodies. The framework regions described herein are determined based on the boundaries of the CDR numbering system. In other words, when the CDRs are determined by, for example, Kabat, IMGT, or Chothia, the framework regions are the amino acid residues surrounding the CDRs of the variable region from N-terminus to C-terminus, i.e., in the format FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. For example, FR1 is defined as the amino acid residues N-terminal to the CDR1 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR2 is defined as the amino acid residues between the CDR1 and CDR2 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; FR3 is defined as the amino acid residues between the CDR2 and CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system; and FR4 is defined as the amino acid residues C-terminal to the CDR3 amino acid residues as defined, for example, by the Kabat numbering system, the IMGT numbering system, or the Chothia numbering system.

[0231] In certain embodiments, the antibodies or antigen-binding fragments thereof described herein comprise an amino acid sequence having a certain percent identity compared to any one of the above antibodies. In some embodiments, an anti-NKp46 antibody is provided that comprises a VH having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 67. In some embodiments, an anti-NKp46 antibody is provided that comprises a VL having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 68. In some embodiments, an anti-NKp46 antibody is provided, comprising a VH having at least about any one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 67; and a VL having at least about any one of 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 68.

[0232] In some embodiments, a VH or VL sequence having at least about 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to a reference sequence contains substitutions (e.g., conservative substitutions), insertions, or deletions compared to the reference sequence, but antibodies containing the sequence retain the ability to bind to NKp46. In some embodiments, a total of 1 to 10 amino acids are substituted, inserted, and / or deleted. In some embodiments, the substitutions, insertions, or deletions occur in regions outside the CDRs (i.e., within the FRs).

[0233] In another aspect, provided herein is an antibody that competes with any of the NKp46 antibodies described herein for binding to NKp46. In another aspect, provided herein is an antibody that binds to the same epitope as any of the NKp46 antibodies described herein. In another aspect, provided is an NKp46 antibody that binds to an epitope on NKp46 that overlaps with the epitope on NKp46 bound by the NKp46 antibodies described herein.

[0234] In one embodiment, an antibody is provided that competes with an NKp46 reference antibody for binding to NKp46. In another embodiment, an NKp46 antibody is provided that binds to the same NKp46 epitope as the NKp46 reference antibody. In another embodiment, an NKp46 antibody is provided that binds to an epitope on NKp46 that overlaps with the epitope on NKp46 bound by the NKp46 reference antibody.

[0235] In one embodiment, the NKp46 reference antibody comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 67, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 68.

[0236] The antibodies provided herein can be from any animal origin, including birds and mammals (e.g., human, monkey, murine, donkey, sheep, rabbit, goat, guinea pig, camel, horse, or chicken). In certain embodiments, the antibodies provided herein are human or humanized monoclonal antibodies. As used herein, a "human" antibody includes antibodies having the amino acid sequence of a human immunoglobulin, including antibodies isolated from a human immunoglobulin library or from a mouse expressing an antibody derived from human genes.

[0237] In certain embodiments, the antibody is a fully murine antibody. In certain embodiments, the antibody is a mouse-human chimeric antibody. In certain embodiments, the antibody is a humanized antibody. In certain embodiments, the antibody is a fully human antibody. In other embodiments, the antibodies provided herein are humanized antibodies (e.g., comprising human constant and framework regions). The antibodies provided herein may be bispecific, trispecific, or of greater multispecificity.

[0238] In some embodiments, the antibodies or antigen-binding fragments provided herein have a K of less than 1000 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 100 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 50 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 40 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 30 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 20 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 10 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 9 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 8 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 7 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 6 nM. DIn some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 5 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 4 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 3 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 2 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 1 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 0.1 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKG2d with a K of less than 0.01 nM. D It binds to NKG2d at K D or K D The K value can also be measured by any method known in the art, for example, using biolayer interferometry (BLI) or surface plasmon resonance (SPR) by Octet® (e.g., using an Octet® Red 96 system) or by Biacore® (e.g., using a Biacore® TM-2000 or Biacore® TM-3000). The "on rate" or "rate of association" or "association rate" or "k" can also be measured using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, for example, using an Octet® Red 96, Biacore® TM-2000, or Biacore® TM-3000 system. In certain embodiments, the K Dis determined by Biacore® assay. In some embodiments, the NKG2d is human NKG2d. In some embodiments, the NKG2d is cynomolgus monkey NKG2d. In some embodiments, the NKG2d is rat NKG2d. In other embodiments, the NKG2d is mouse NKG2d.

[0239] In other embodiments, the antibodies or antigen-binding fragments provided herein have a K of less than 1000 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 100 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 50 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 40 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 30 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 20 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 10 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 9 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 8 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 7 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 6 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 5 nM. DIn some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 4 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 3 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 2 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 1 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 0.1 nM. D In some embodiments, the antibodies or antigen-binding fragments provided herein bind to NKp46 with a K of less than 0.01 nM. D It binds to NKp46 at K D or K D The K value can also be measured by any method known in the art, for example, using biolayer interferometry (BLI) or surface plasmon resonance (SPR) by Octet® (e.g., using an Octet® Red 96 system) or Biacore® (e.g., using a Biacore® TM-2000 or Biacore® TM-3000). Also, the "on rate" or "rate of association" or "association rate" or "k" can be measured using the same biolayer interferometry (BLI) or surface plasmon resonance (SPR) techniques described above, for example, using an Octet® Red 96, Biacore® TM-2000, or Biacore® TM-3000 system. In certain embodiments, the K D is determined by Biacore® assay. In some embodiments, the NKp46 is human NKp46. In some embodiments, the NKp46 is cynomolgus monkey NKp46. In some embodiments, the NKp46 is rat NKp46. In other embodiments, the NKp46 is mouse NKp46.

[0240] In some embodiments, provided herein are antibodies that specifically bind to NKG2d and can modulate NK cell activity. In some embodiments, provided herein are antibodies that specifically bind to NKG2d and can modulate NKG2d-expressing immune effector cell activity.

[0241] In some embodiments, provided herein are antibodies that specifically bind to NKp46 and are capable of modulating NK cell activity.

[0242] In some embodiments, provided herein are antibodies that specifically bind to NKp46 and can regulate the activity of NKp46-expressing immune effector cells. In some embodiments, the NKp46-expressing immune effector cells are T cells. In some embodiments, the T cells are gamma delta T cells. In some embodiments, the T cells are a mucosal population of innate lymphoid cells.

[0243] In some embodiments, provided herein are antibodies that specifically bind to NKp46 and can regulate T cell activity. In some embodiments, the T cells are gamma delta T cells. In some embodiments, the T cells are a mucosal population of innate lymphoid cells.

[0244] In some embodiments, the antibodies described herein are capable of activating NK cells. In some embodiments, the antibodies described herein activate NK cell activity by at least about 10%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 20%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 30%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 40%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 50%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 60%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 70%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 80%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 90%. In some embodiments, the antibodies described herein activate NK cell activity by at least about 95%. In certain embodiments, the antibodies described herein activate NK cell activity by at least about 15% to about 65%. In certain embodiments, the antibodies described herein activate NK cell activity by at least about 20% to about 65%. In certain embodiments, the antibodies described herein activate NK cell activity by at least about 30% to about 65%. In certain embodiments, the NK cells are human NK cells.

[0245] In some embodiments, the antibodies described herein can promote IFNg production by NK cells. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 10%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 20%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 30%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 40%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 50%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 60%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 70%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 80%. In some embodiments, the antibodies described herein promote IFNg production by NK cells by at least 90%. In some embodiments, the antibodies described herein stimulate IFNg production by NK cells by at least 95%. In certain embodiments, the antibodies described herein stimulate IFNg production by NK cells by at least about 15% to about 65%. In certain embodiments, the antibodies described herein stimulate IFNg production by NK cells by at least about 20% to about 65%. In certain embodiments, the antibodies described herein stimulate IFNg production by NK cells by at least about 30% to about 65%. In certain embodiments, the NK cells are human NK cells.

[0246] multispecific molecules The multispecific molecules provided herein comprise a binding domain capable of binding to an antigen present on an NK cell. In some embodiments, the antigen is NKG2d. In some embodiments, the antigen is NKp46. In some embodiments, the first binding domain is as described or derived from an antibody described above.

[0247] In addition to the above domains, the multispecific molecules provided herein comprise an additional domain capable of binding to a second antigen. In some embodiments, the second binding domain is capable of binding to an antigen expressed on a tumor cell. In some embodiments, the second binding domain is capable of binding to a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA). In some embodiments, the second binding domain is capable of binding to BCMA. In some embodiments, the second binding domain is capable of binding to GPRC5d.

[0248] Tumor antigens are proteins produced by tumor cells that can elicit an immune response, particularly a T-cell mediated immune response. Exemplary tumor antigens include glioma-associated antigen, carcinoembryonic antigen (CEA), β-human chorionic gonadotropin, alphafetoprotein (AFP), lectin-reactive AFP, thyroglobulin, RAGE-1, MN-CAIX, human telomerase reverse transcriptase, RU1, RU2 (AS), intestinal carboxylesterase, mut hsp70-2, M-CSF, prostase, prostate-specific antigen (PSA), PAP, NY-ESO-1, LAGE-1a, p53, prostein, PSMA, HER2 / neu, survivin, and telomerase, prostate-carcinoma tumor antigen-1 (PCTA-1), MAGE, ELF2M, neutrophil elastase, ephrin B2, insulin growth factor (IGF), and telomerase. These include, but are not limited to, IGF-I, IGF-II, IGF-I receptor, and mesothelin.

[0249] In some embodiments, the tumor antigen comprises one or more antigenic cancer epitopes associated with malignant tumors. Malignant tumors express numerous proteins that can serve as target antigens for immune attack. These molecules include, but are not limited to, tissue-specific antigens such as MART-1, tyrosinase, and gp100 in melanoma, and prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA) in prostate cancer. Other target molecules belong to the group of transformation-related molecules, such as the oncogene HER2 / Neu / ErbB-2. Yet another group of target antigens are oncofetal antigens, such as carcinoembryonic antigen (CEA).

[0250] In some embodiments, the tumor antigen is a tumor-specific antigen (TSA) or tumor-associated antigen (TAA). TSAs are unique to tumor cells and are not present on other cells in the body. TAA-associated antigens are not unique to tumor cells, but are instead expressed on normal cells under conditions that cannot induce a state of immune tolerance to the antigen. Expression of an antigen on a tumor can occur under conditions that allow the immune system to respond to the antigen. TAAs can be antigens that are expressed on normal cells during fetal development, when the immune system is immature and unable to respond, or they can be antigens that are normally present at very low levels on normal cells but are expressed at much higher levels on tumor cells.

[0251] Non-limiting examples of TSA or TAA antigens include differentiation antigens such as MART-1 / MelanA (MART-I), gp100 (Pmel 17), tyrosinase, TRP-1, and TRP-2, and tumor-specific multilineage antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, and p15; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutated tumor suppressor genes such as p53, Ras, and HER2 / neu; unique tumor antigens resulting from chromosomal translocations; BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, and MYL-RAR; and viral antigens, such as Epstein-Barr virus antigen EBVA and human papillomavirus (HPV) antigens E6 and E7.

[0252] In some embodiments, the multispecific molecules provided herein are multispecific antibodies. The antibodies provided herein include, but are not limited to, synthetic antibodies, monoclonal antibodies, recombinantly produced antibodies, human antibodies, humanized antibodies, chimeric antibodies, etc.

[0253] In particular, antibodies provided herein include immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen-binding site that immunospecifically binds to an antigen. The immunoglobulin molecules provided herein can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass of immunoglobulin molecule. In some embodiments, the antibody is an IgG antibody. In some embodiments, the IgG antibody is an IgG1 antibody. In some embodiments, the IgG antibody is an IgG2, IgG3, or IgG4 antibody.

[0254] In some embodiments of the various multispecific molecules provided herein, the first binding domain and / or the second binding domain are antibody variants and / or derivatives, including antibody fragments, that retain the ability to specifically bind to an epitope. In other embodiments of the various multispecific molecules provided herein, the first binding domain and / or the second binding domain are antibody variants and / or derivatives, including antibody fragments, that retain the ability to specifically bind to an epitope.Exemplary fragments include Fab fragments (antibody fragments comprising a portion of a light chain and a heavy chain cross-linked by a disulfide bond, including the antigen-binding domain), Fab' (antibody fragments comprising a single anti-binding domain comprising a Fab and an additional portion of the heavy chain through the hinge region), F(ab')2 (two Fab' molecules linked by an interchain disulfide bond at the hinge region of the heavy chain, which Fab' molecules may be directed to the same or different epitopes), bispecific Fab (a Fab molecule with two antigen-binding domains, each of which may be directed to a different epitope), single-chain Fab chains containing variable regions, also known as scFvs (the variable antigen-binding determining regions of a single light and heavy chain of an antibody linked together by a chain of 10-25 amino acids), disulfide-linked Fvs, or dsFvs (the variable antigen-binding determining regions of a single light and heavy chain of an antibody linked together by a disulfide bond), camelized VH (in which some amino acids at the VH interface are found in the heavy chain of a native camelid antibody). , the variable antigen-binding determining region of a single heavy chain of an antibody), bispecific scFvs (scFv or dsFv molecules with two antigen-binding domains, each of which may be directed against a different epitope), diabodies (dimerized scFvs formed when the VH domain of a first scFv associates with the VL domain of a second scFv and the VL domain of the first scFv associates with the VH domain of a second scFv; the two antigen-binding regions of the diabody may be directed against the same or different epitopes), triabodies (trimerized scFvs, formed in a manner similar to diabodies, but with three antigen-binding domains created in a single complex, which may be directed against the same or different epitopes), and tetrabodies (tetramerized scFvs, formed in a manner similar to diabodies, but with four antigen-binding domains created in a single complex, which may be directed against the same or different epitopes). Derivatives of antibodies also include one or more CDR sequences of the antibody binding site. The CDR sequences may be linked together on a scaffold when two or more CDR sequences are present. In certain embodiments, the antibodies provided herein comprise a single-chain Fv ("scFv").An scFv is an antibody fragment comprising the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Generally, the scFv polypeptide further comprises a polypeptide linker between the VH and VL domains which enables the scFv to form the desired structure for antigen binding. For a review of scFvs, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds. Springer-Verlag, New York, pp. 269-315 (1994).

[0255] In certain embodiments, the antibody that binds to NKG2d comprises a VH region and a VL region. In some embodiments, the NKG2d antibody is a single-chain antibody. In some embodiments, the NKG2d antibody is a single-domain antibody. In some embodiments, the NKG2d antibody is a nanobody. In certain embodiments, the NKG2d antibody is a VHH antibody. In certain embodiments, the NKG2d antibody is a llama antibody. In some embodiments, the NKG2d antibody is not a single-chain antibody. In some embodiments, the NKG2d antibody is not a single-domain antibody. In some embodiments, the NKG2d antibody is not a nanobody. In certain embodiments, the NKG2d antibody is not a VHH antibody. In certain embodiments, the NKG2d antibody is not a llama antibody. In some embodiments, the NKG2d antibody is a multispecific antibody. In other embodiments, the NKG2d antibody is a bispecific antibody. In certain embodiments, the multispecific antibody comprises an antigen-binding fragment of an NKG2d antibody provided herein. In other embodiments, the bispecific antibody comprises an antigen-binding fragment of an NKG2d antibody provided herein.

[0256] In another aspect, provided herein are multispecific antibodies that bind to NKG2d. In some embodiments, the multispecific antibody is a bispecific antibody. In some embodiments, the multispecific antibody is a trispecific antibody. In some embodiments, the multispecific antibody is a tetraspecific antibody. In one embodiment, the multispecific NKG2d antibody comprises (a) a first binding domain that binds to NKG2d, and (b) a second binding domain that binds to a second target. In one embodiment, the multispecific NKG2d antibody comprises (a) a first binding domain that binds to NKG2d, (b) a second binding domain that binds to a second target, and (c) a third binding domain that binds to a third target. In one embodiment, the multispecific NKG2d antibody comprises (a) a first binding domain that binds to NKG2d, (b) a second binding domain that binds to a second target, (c) a third binding domain that binds to a third target, and (d) a fourth binding domain that binds to a fourth target.

[0257] In certain embodiments, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to a second target that is not NKG2d. In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to a second target that binds to an antigen expressed on tumor cells. In some embodiments, the second binding domain binds to BCMA. In some embodiments, the second binding domain binds to GPRC5d.

[0258] In certain embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VH region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VH region of any one of the antibodies described herein and the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described. In some embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKG2d bispecific antibodies comprising a binding domain that binds to NKG2d having the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein and the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein.

[0259] In certain embodiments, the anti-NKG2d antibody is a bispecific antibody. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VH region of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VL region of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VH region of the anti-BCMA antibodies provided herein and the VL region of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VH CDR1, VH CDR2, and VH CDR3 of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VL CDR1, VL CDR2, and VL CDR3 of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to BCMA having the VH CDR1, VH CDR2, and VH CDR3 of the anti-BCMA antibodies provided herein and the VL CDR1, VL CDR2, and VL CDR3 of the anti-BCMA antibodies provided herein.

[0260] In certain embodiments, the anti-NKG2d antibody is a bispecific antibody. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH region of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VL region of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH region of an anti-GPRC5d antibody provided herein and the VL region of an anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH CDR1, VH CDR2, and VH CDR3 of an anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VL CDR1, VL CDR2, and VL CDR3 of an anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKG2d bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH CDR1, VH CDR2, and VH CDR3 of an anti-GPRC5d antibody provided herein and the VL CDR1, VL CDR2, and VL CDR3 of an anti-GPRC5d antibody provided herein.

[0261] In some embodiments, the first binding domain that binds to NKG2d is as described or derived from an antibody described above. In some particular embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 4, 5, and 6, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 7, 8, and 9, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 10, 11, and 12, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 13, 14, and 15, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 16, 17, and 18, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 19, 20, and 21, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 22, 23, and 24, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 25, 26, and 27, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 28, 29, and 30, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 31, 32, and 33, respectively.In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 2. In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL that includes a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 3. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 2, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 3. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 2. In some embodiments, the first binding domain that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 2 and a VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 2. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 3.In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO:2, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO:3.

[0262] In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 36, 37, and 38, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 39, 40, and 41, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 42, 43, and 44, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 45, 46, and 47, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 48, 49, and 50, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 51, 52, and 53, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 54, 55, and 56, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 57, 58, and 59, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 60, 61, and 62, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 63, 64, and 65, respectively.In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 34. In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL that includes a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 35. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 34, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 35. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 34. In some embodiments of the multispecific antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 35.In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 34, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 35.

[0263] In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Kabat numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Chothia numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKG2d are according to the AbM numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Contact numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the IMGT numbering system.

[0264] In some embodiments of the multispecific NKG2d antibodies provided herein, the first binding domain binds to an NKG2d antigen. In some embodiments, the first binding domain binds to an NKG2d epitope. In some embodiments, the first binding domain specifically binds to NKG2d. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an NKG2d antigen. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an NKG2d epitope. In some embodiments, NKG2d is present on the surface of NK cells.

[0265] In another aspect, provided herein are multispecific antibodies that compete for binding to NKG2d with any of the NKG2d antibodies described herein. In another aspect, provided herein are multispecific antibodies that bind to the same epitope as any of the NKG2d antibodies described herein. In another aspect, provided herein are multispecific NKG2d antibodies that bind to an epitope on NKG2d that overlaps with the epitope on NKG2d bound by the NKG2d antibodies described herein.

[0266] In one embodiment, multispecific antibodies are provided that compete with an NKG2d reference antibody for binding to NKG2d. In another embodiment, multispecific NKG2d antibodies are provided that bind to the same NKG2d epitope as the NKG2d reference antibody. In another embodiment, multispecific NKG2d antibodies are provided that bind to an epitope on NKG2d that overlaps with the epitope on NKG2d bound by the NKG2d reference antibody.

[0267] In one embodiment, the NKG2d reference antibody comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 2, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 3. In one embodiment, the NKG2d reference antibody comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 34, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 35.

[0268] In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the third target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the fourth target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d antigen and the third target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d antigen and the fourth target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the third target is not an NKG2d antigen and the fourth target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d antigen, the third target is not an NKG2d antigen, and the fourth target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d antibodies provided herein, the third target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d antibodies provided herein, the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d epitope and the third target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d epitope and the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d antibodies provided herein, the third target is not an NKG2d epitope and the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is not an NKG2d epitope, the third target is not an NKG2d epitope, and the fourth target is not an NKG2d epitope.In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is BCMA. In some embodiments of the multispecific NKG2d antibodies provided herein, the second target is GPRC5d.

[0269] Binding of a multispecific antibody provided herein to NKG2d present on the surface of an NK cell and binding of a second target antigen present on the surface of a second target cell can, for example, result in killing of the second target cell. In other embodiments, binding of a multispecific antibody provided herein to NKG2d present on the surface of an NK cell and binding of a second target antigen can, for example, result in activation of the NK cell. In some embodiments, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to a cancer antigen present on the surface of a cancer cell. In some embodiments, the antigen on the surface of the cancer cell is a tumor-specific antigen. In some embodiments, the antigen on the surface of the cancer cell is a tumor-associated antigen. In some embodiments, the antigen on the surface of the cancer cell is a neoantigen. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKG2d. In some embodiments, NKG2d is present on the surface of an NK cell. In some embodiments, when the bispecific antibody binds to NKG2d on the surface of an NK cell and an antigen on the surface of the cancer cell, the cancer cell is killed.

[0270] In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to BCMA. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKG2d. In some embodiments, NKG2d is present on the surface of an NK cell. In some embodiments, BCMA is present on the surface of a cell. In certain embodiments, NKG2d is present on the surface of an NK cell and BCMA is present on the surface of a cell. In some embodiments, when the bispecific antibody binds to NKG2d on the surface of an NK cell and BCMA on the surface of a cell, the cell bearing BCMA on its surface is killed. In some embodiments, BCMA is present on the surface of a cancer cell. In certain embodiments, NKG2d is present on the surface of an NK cell and BCMA is present on the surface of a cancer cell. In some embodiments, when the bispecific antibody binds to NKG2d on the surface of an NK cell and BCMA on the surface of a cancer cell, the cancer cell is killed. In certain embodiments, bispecific antibodies are contemplated that comprise any of the NKG2d antibodies provided herein as a first binding domain. In addition, in certain embodiments, bispecific antibodies are also contemplated that comprise any of the NKG2d antibodies provided herein as a first binding domain and a second binding domain that binds to BCMA.

[0271] In some embodiments, the multispecific antibodies provided herein are bispecific antibodies comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to BCMA, wherein the second binding domain that binds to BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 101, 102, and 103, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 104, 105, and 106, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 107, 108, and 109, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 110, 111, and 112, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 113, 114, and 115, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 116, 117, and 118, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 119, 120, and 121, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 122, 123, and 124, respectively.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 125, 126, and 127, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 128, 129, and 130, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VL having the amino acid sequence of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99 and a VL having the amino acid sequence of SEQ ID NO: 100.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 99. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 99, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 100.

[0272] In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to GPRC5d. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKG2d. In some embodiments, NKG2d is present on the surface of an NK cell. In some embodiments, GPRC5d is present on the surface of a cell. In certain embodiments, NKG2d is present on the surface of an NK cell and GPRC5d is present on the surface of a cell. In some embodiments, when the bispecific antibody binds to NKG2d on the surface of an NK cell and GPRC5d on the surface of a cell, the cell bearing GPRC5d on its surface is killed. In some embodiments, GPRC5d is present on the surface of a cancer cell. In certain embodiments, NKG2d is present on the surface of an NK cell and GPRC5d is present on the surface of a cancer cell. In some embodiments, when the bispecific antibody binds to NKG2d on the surface of an NK cell and GPRC5d on the surface of a cancer cell, the cancer cell is killed. In certain embodiments, bispecific antibodies are contemplated that comprise any of the NKG2d antibodies provided herein as a first binding domain. In addition, in certain embodiments, bispecific antibodies are also contemplated that comprise any of the NKG2d antibodies provided herein as a first binding domain and a second binding domain that binds to GPRC5d.

[0273] In some embodiments, the multispecific antibody provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to GPRC5d, wherein the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 133, 134, and 135, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 136, 137, and 138, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 139, 140, and 141, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 142, 143, and 144, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 145, 146, and 147, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 148, 149, and 150, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 151, 152, and 153, respectively; and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 154, 155, and 156, respectively.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 157, 158, and 159, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 160, 161, and 162, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 131. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 131, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH having the amino acid sequence of SEQ ID NO: 131. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL having the amino acid sequence of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH having the amino acid sequence of SEQ ID NO: 131 and a VL having the amino acid sequence of SEQ ID NO: 132.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 131. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 131, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 132.

[0274] In certain embodiments, multispecific antibodies are provided that comprise the NKG2d antibodies provided herein in a knob-in-hole format. In certain embodiments, bispecific antibodies are provided that comprise the NKG2d antibodies provided herein in a knob-in-hole format. In certain embodiments, trispecific antibodies are provided that comprise the NKG2d antibodies provided herein in a knob-in-hole format. In certain embodiments, tetraspecific antibodies are provided that comprise the NKG2d antibodies provided herein in a knob-in-hole format. Other specificities can be added to antibodies in a knob-in-hole format (e.g., adding an scFv to the N- or C-terminus) using methods well known in the art. Furthermore, other formats and methods of making multispecific antibodies are also known in the art and are contemplated. In some embodiments, the NKG2d antibodies provided herein are included in bispecific antibodies. In some embodiments, the NKG2d antibodies provided herein are included in trispecific antibodies. In some embodiments, the NKG2d antibodies provided herein are included in tetraspecific antibodies. In some embodiments, the NKG2d bispecific antibodies provided herein are included in multispecific antibodies.

[0275] In certain embodiments, a multispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d epitope and a second binding domain that binds to a second epitope, wherein the first NKG2d epitope and the second epitope are not the same. In certain embodiments, a bispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d epitope and a second binding domain that binds to a second epitope, wherein the first NKG2d epitope and the second epitope are not the same. In certain embodiments, a trispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d epitope, a second binding domain that binds to a second epitope, and a third binding domain that binds to a third epitope, wherein the first NKG2d epitope, the second epitope, and the third epitope are not the same. In certain embodiments, a tetraspecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d epitope, a second binding domain that binds to a second epitope, a third binding domain that binds to a third epitope, and a fourth binding domain that binds to a fourth epitope, wherein the first NKG2d epitope, the second epitope, the third epitope, and the fourth epitope are not the same. In certain embodiments, a multispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d antigen and a second binding domain that binds to a second antigen, wherein the first NKG2d antigen and the second antigen are not the same. In certain embodiments, a bispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d antigen and a second binding domain that binds to a second antigen, wherein the first NKG2d antigen and the second antigen are not the same.In certain embodiments, a trispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d antigen, a second binding domain that binds to a second antigen, and a third binding domain that binds to a third antigen, wherein the first NKG2d antigen, the second antigen, and the third antigen are not the same. In certain embodiments, a tetraspecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to a first NKG2d antigen, a second binding domain that binds to a second antigen, a third binding domain that binds to a third antigen, and a fourth binding domain that binds to a fourth antigen, wherein the first NKG2d antigen, the second antigen, the third antigen, and the fourth antigen are not the same. In certain embodiments, an NKG2d antibody or antigen-binding fragment thereof provided herein specifically binds to NKG2d.

[0276] In some embodiments, the multispecific antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region, and the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKG2d antibody is not a single-domain antibody or a nanobody. In some embodiments, the third binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the fourth binding domain comprises a heavy chain variable region and a light chain variable region.

[0277] In certain embodiments, an NKG2d multispecific antibody or antigen-binding fragment thereof binds to a first epitope located on NKG2d and a second epitope on a second target antigen. Provided herein in some embodiments is a multispecific antibody comprising (a) a first binding domain that binds to an NKG2d antigen and (b) a second binding domain that binds to a second target antigen. Provided herein in some embodiments is a multispecific antibody comprising (a) a first binding domain that specifically binds to an NKG2d antigen and (b) a second binding domain that specifically binds to a second target antigen. Provided herein in some embodiments is a multispecific antibody comprising (a) a first binding domain that binds to a first epitope on an NKG2d antigen and (b) a second binding domain that binds to a second epitope on a second target antigen. In some embodiments, provided herein are multispecific antibodies comprising: (a) a first binding domain that specifically binds to a first epitope on an NKG2d antigen; and (b) a second binding domain that specifically binds to a second epitope on a second target antigen.

[0278] In certain embodiments, the NKG2d antigen is present on the surface of an NK cell. In certain embodiments, the second target antigen is not NKG2d. Binding of an NKG2d multispecific antibody to NKG2d present on the surface of an NK cell and binding of a second target antigen present on the surface of a second target cell can, for example, result in killing of the second target cell. In other embodiments, binding of an NKG2d multispecific antibody to NKG2d present on the surface of an NK cell and binding of a second target antigen can, for example, result in activation of the NK cell.

[0279] In another aspect, provided herein is a multispecific antibody comprising a first binding domain that binds NKG2d and a second binding domain that binds BCMA (a "multispecific NKG2d / BCMA antibody"). In some embodiments, the multispecific NKG2d / BCMA antibody is a bispecific antibody. In some embodiments, the multispecific NKG2d / BCMA antibody is a trispecific antibody. In some embodiments, the multispecific NKG2d / BCMA antibody is a tetraspecific antibody.

[0280] In some embodiments, the multispecific NKG2d / BCMA antibodies provided herein comprise (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to BCMA. In one embodiment, the multispecific NKG2d / BCMA antibodies comprise (a) a first binding domain that binds to NKG2d, (b) a second binding domain that binds to BCMA, and (c) a third binding domain that binds to a third target. In one embodiment, the multispecific NKG2d / BCMA antibodies comprise (a) a first binding domain that binds to NKG2d, (b) a second binding domain that binds to BCMA, (c) a third binding domain that binds to a third target, and (d) a fourth binding domain that binds to a fourth target.

[0281] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of the VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 2. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of the VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 2, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 2. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO:2 and a VL having the amino acid sequence of SEQ ID NO:3.

[0282] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of a VH having the amino acid sequence of SEQ ID NO: 34. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 34, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO: 35.

[0283] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Kabat numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Chothia numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKG2d are according to the AbM numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Contact numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKG2d are according to the IMGT numbering system.

[0284] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain binds to an NKG2d antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain binds to an NKG2d epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain specifically binds to NKG2d. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form the binding site for an NKG2d antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an epitope of NKG2d. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, NKG2d is present on the surface of NK cells.

[0285] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VL that includes a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VL having the amino acid sequence of SEQ ID NO: 100. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99 and a VL having the amino acid sequence of SEQ ID NO: 100.

[0286] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the Kabat numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the Chothia numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the AbM numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the Contact numbering system. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the IMGT numbering system.

[0287] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain binds to a BCMA antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain binds to a BCMA epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain specifically binds to BCMA. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain form a binding site for an antigen of BCMA. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain form a binding site for an epitope of BCMA. In some embodiments, BCMA is present on the surface of a tumor cell.

[0288] In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the fourth target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not an NKG2d antigen and the fourth target is not an NKG2d antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not a BCMA antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the fourth target is not a BCMA antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not a BCMA antigen and the fourth target is not a BCMA antigen. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not an NKG2d epitope and the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not a BCMA epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the fourth target is not a BCMA epitope. In some embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the third target is not a BCMA epitope and the fourth target is not a BCMA epitope.

[0289] In certain embodiments, the target is derived from a mammal. In certain embodiments, the target is derived from a rat. In certain embodiments, the target is derived from a mouse. In certain embodiments, the target is derived from a primate. In certain embodiments, the target is derived from a human.

[0290] In certain embodiments, multispecific NKG2d / BCMA antibodies in a knobs-in-hole format are provided. In certain embodiments, bispecific NKG2d / BCMA antibodies in a knobs-in-hole format are provided. In certain embodiments, trispecific antibodies in a knobs-in-hole format are provided. In certain embodiments, tetraspecific antibodies in a knobs-in-hole format are provided. Other specificities can be added to antibodies in a knobs-in-hole format (e.g., adding an scFv to the N- or C-terminus) using methods well known in the art. Furthermore, other formats and methods of making multispecific antibodies are also known in the art and are contemplated. In some embodiments, the NKG2d / BCMA antibodies provided herein are included in bispecific antibodies. In some embodiments, the NKG2d / BCMA antibodies provided herein are included in trispecific antibodies. In some embodiments, the NKG2d / BCMA antibodies provided herein can be included in tetraspecific antibodies. In some embodiments, the NKG2d / BCMA bispecific antibodies provided herein are included in multispecific antibodies.

[0291] In certain embodiments, the trispecific NKG2d / BCMA antibodies provided herein comprise a first binding domain comprising an NKG2d antibody provided herein that binds to an NKG2d epitope, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA epitope, and a third binding domain that binds to a third epitope, wherein the NKG2d epitope, the BCMA epitope, and the third epitope are not the same. In certain embodiments, a tetraspecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to an NKG2d epitope, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA epitope, a third binding domain that binds to a third epitope, and a fourth binding domain that binds to a fourth epitope, wherein the NKG2d epitope, the BCMA epitope, the third epitope, and the fourth epitope are not the same. In certain embodiments, a trispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to an NKG2d antigen, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA antigen, and a third binding domain that binds to a third antigen, wherein the NKG2d antigen, the BCMA antigen, and the third antigen are not the same. In certain embodiments, the tetraspecific antibodies provided herein bind to an NKG2d antigen, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA antigen, a third binding domain that binds to a third antigen, a fourth binding domain that binds to a fourth antigen, The NKG2d antigen, the BCMA antigen, the third antigen, and the fourth antigen are not the same. In certain embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds NKG2d specifically binds to NKG2d. In other embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the second binding domain that binds BCMA specifically binds to BCMA. In yet other embodiments of the multispecific NKG2d / BCMA antibodies provided herein, the first binding domain that binds NKG2d specifically binds to NKG2d and the second binding domain that binds BCMA specifically binds to BCMA.

[0292] In some embodiments, the multispecific NKG2d / BCMA antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region, and the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKG2d antibody is not a single-domain antibody or a nanobody. In some embodiments, the third binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the fourth binding domain comprises a heavy chain variable region and a light chain variable region.

[0293] In certain embodiments, an NKG2d / BCMA multispecific antibody or antigen-binding fragment thereof binds to a first epitope located on NKG2d and a second epitope located on BCMA. Provided herein in some embodiments are multispecific NKG2d / BCMA antibodies comprising (a) a first binding domain that binds to the NKG2d antigen and (b) a second binding domain that binds to the BCMA antigen. Provided herein in some embodiments are multispecific NKG2d / BCMA antibodies comprising (a) a first binding domain that specifically binds to the NKG2d antigen and (b) a second binding domain that specifically binds to the BCMA antigen. Provided herein in some embodiments are multispecific NKG2d / BCMA antibodies comprising (a) a first binding domain that binds to a first epitope on the NKG2d antigen and (b) a second binding domain that binds to a second epitope on the BCMA antigen. In some embodiments, provided herein are multispecific antibodies comprising: (a) a first binding domain that specifically binds to a first epitope on the NKG2d antigen; and (b) a second binding domain that specifically binds to a second epitope on the BCMA antigen.

[0294] In certain embodiments, the NKG2d antigen is on the surface of an NK cell. In certain embodiments, the BCMA antigen is on the surface of a tumor cell. Binding of an NKG2d / BCMA multispecific antibody to NKG2d present on the surface of an NK cell and BCMA present on the surface of a tumor cell can result in, for example, tumor cell killing. In other embodiments, binding of an NKG2d / BCMA multispecific antibody to NKG2d present on the surface of an NK cell and BCMA present on the surface of a tumor cell can result in, for example, NK cell activation.

[0295] In another aspect, provided herein is a multispecific antibody comprising a first binding domain that binds to NKG2d and a second binding domain that binds to GPRC5d (a "multispecific NKG2d / GPRC5d antibody"). In some embodiments, the multispecific NKG2d / GPRC5d antibody is a bispecific antibody. In some embodiments, the multispecific NKG2d / GPRC5d antibody is a trispecific antibody. In some embodiments, the multispecific NKG2d / GPRC5d antibody is a tetraspecific antibody.

[0296] In one embodiment, the multispecific NKG2d / GPRC5d antibody comprises (a) a first binding domain that binds to NKG2d and (b) a second binding domain that binds to GPRC5d. In one embodiment, the multispecific NKG2d / GPRC5d antibody comprises (a) a first binding domain that binds to NKG2d, (b) a second binding domain that binds to GPRC5d, and (c) a third binding domain that binds to a third target. In one embodiment, the multispecific NKG2d / GPRC5d antibody comprises (a) a first binding domain that binds to NKG2d, (b) a second binding domain that binds to GPRC5d, (c) a third binding domain that binds to a third target, and (d) a fourth binding domain that binds to a fourth target.

[0297] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of a VH having the amino acid sequence of SEQ ID NO: 2. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of a VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 2, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 2. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 3. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 2 and a VL having the amino acid sequence of SEQ ID NO: 3.

[0298] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, a first binding domain that binds to NKG2d comprises a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of a VH having the amino acid sequence of SEQ ID NO: 34. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, a first binding domain that binds to NKG2d comprises a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 34, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises a VL having the amino acid sequence of SEQ ID NO: 35. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d comprises a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO: 35.

[0299] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Kabat numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Chothia numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKG2d are according to the AbM numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the Contact numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKG2d are according to the IMGT numbering system.

[0300] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain binds to an NKG2d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain binds to an NKG2d epitope. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain specifically binds to NKG2d. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form the binding site for an NKG2d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an epitope of NKG2d. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, NKG2d is present on the surface of NK cells.

[0301] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 131. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL that includes a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 132. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 131, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 132. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH having the amino acid sequence of SEQ ID NO: 131. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL having the amino acid sequence of SEQ ID NO: 132. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH having the amino acid sequence of SEQ ID NO: 131 and a VL having the amino acid sequence of SEQ ID NO: 132.

[0302] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds GPRC5d are according to the Kabat numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds GPRC5d are according to the Chothia numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds GPRC5d are according to the AbM numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds GPRC5d are according to the Contact numbering system. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds GPRC5d are according to the IMGT numbering system.

[0303] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain binds to the GPRC5d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain binds to a GPRC5d epitope. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain specifically binds to GPRC5d. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain form a binding site for the GPRC5d antigen. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain form a binding site for the GPRC5d epitope. In some embodiments, GPRC5d is present on the surface of tumor cells.

[0304] In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not the NKG2d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the fourth target is not the NKG2d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not the NKG2d antigen and the fourth target is not the NKG2d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not the GPRC5d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the fourth target is not the GPRC5d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not the GPRC5d antigen and the fourth target is not the GPRC5d antigen. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not an NKG2d epitope and the fourth target is not an NKG2d epitope. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the third target is not a GPRC5d epitope. In some embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the fourth target is not a GPRC5d epitope. In some embodiments of the multispecific NKG2d GPRC5d antibodies provided herein, the third target is not a GPRC5d epitope and the fourth target is not a GPRC5d epitope.

[0305] In certain embodiments, the target is derived from a mammal. In certain embodiments, the target is derived from a rat. In certain embodiments, the target is derived from a mouse. In certain embodiments, the target is derived from a primate. In certain embodiments, the target is derived from a human.

[0306] In certain embodiments, multispecific NKG2d / GPRC5d antibodies in a knobs-in-hole format are provided. In certain embodiments, bispecific NKG2d / GPRC5d antibodies in a knobs-in-hole format are provided. In specific embodiments, trispecific antibodies in a knobs-in-hole format are provided. In specific embodiments, tetraspecific antibodies in a knobs-in-hole format are provided. Other specificities can be added to antibodies in a knobs-in-hole format (e.g., adding an scFv to the N- or C-terminus) using methods well known in the art. Furthermore, other formats and methods of making multispecific antibodies are also known in the art and are contemplated. In some embodiments, the NKG2d / GPRC5d antibodies provided herein are comprised in a bispecific antibody. In some embodiments, the NKG2d / GPRC5d antibodies provided herein are comprised in a trispecific antibody. In some embodiments, the NKG2d / GPRC5d antibodies provided herein are comprised in a tetraspecific antibody. In some embodiments, the NKG2d / GPRC5d bispecific antibodies provided herein are comprised in a multispecific antibody.

[0307] In certain embodiments, the trispecific NKG2d / GPRC5d antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to an NKG2d epitope, a second binding domain comprising a GPRC5d antibody provided herein that binds to a GPRC5d epitope, and a third binding domain that binds to a third epitope, wherein the NKG2d epitope, the GPRC5d epitope, and the third epitope are not the same. In certain embodiments, a tetraspecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to an NKG2d epitope, a second binding domain comprising a GPRC5d antibody provided herein that binds to a GPRC5d epitope, a third binding domain that binds to a third epitope, and a fourth binding domain that binds to a fourth epitope, wherein the NKG2d epitope, the GPRC5d epitope, the third epitope, and the fourth epitope are not the same. In certain embodiments, a trispecific antibody provided herein comprises a first binding domain comprising an NKG2d antibody provided herein that binds to an NKG2d antigen, a second binding domain comprising a GPRC5d antibody provided herein that binds to a GPRC5d antigen, and a third binding domain that binds to a third antigen, wherein the NKG2d antigen, the GPRC5d antigen, and the third antigen are not the same. In certain embodiments, the tetraspecific antibodies provided herein bind to an NKG2d antigen, a second binding domain comprising a GPRC5d antibody provided herein that binds to the GPRC5d antigen, a third binding domain that binds to a third antigen, and a fourth binding domain that binds to a fourth antigen, wherein the NKG2d antigen, the GPRC5d antigen, the third antigen, and the fourth antigen are not the same. In certain embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d specifically binds to NKG2d. In other embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the second binding domain that binds to GPRC5d specifically binds to GPRC5d.In still other embodiments of the multispecific NKG2d / GPRC5d antibodies provided herein, the first binding domain that binds to NKG2d specifically binds to NKG2d, and the second binding domain that binds to GPRC5d specifically binds to GPRC5d.

[0308] In some embodiments, the multispecific NKG2d / GPRC5d antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region, and the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKG2d antibody is not a single-domain antibody or a nanobody. In some embodiments, the third binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the fourth binding domain comprises a heavy chain variable region and a light chain variable region.

[0309] In certain embodiments, an NKG2d / GPRC5d multispecific antibody or antigen-binding fragment thereof binds to a first epitope located on NKG2d and a second epitope located on GPRC5d. In some embodiments, provided herein are multispecific NKG2d / GPRC5d antibodies comprising (a) a first binding domain that binds to the NKG2d antigen and (b) a second binding domain that binds to the GPRC5d antigen. In some embodiments, provided herein are multispecific NKG2d / GPRC5d antibodies comprising (a) a first binding domain that specifically binds to the NKG2d antigen and (b) a second binding domain that specifically binds to the GPRC5d antigen. In some embodiments, provided herein are multispecific NKG2d / GPRC5d antibodies comprising (a) a first binding domain that binds to a first epitope on the NKG2d antigen and (b) a second binding domain that binds to a second epitope on the GPRC5d antigen. In some embodiments, provided herein are multispecific antibodies comprising (a) a first binding domain that specifically binds to a first epitope on the NKG2d antigen, and (b) a second binding domain that specifically binds to a second epitope on the GPRC5d antigen.

[0310] In certain embodiments, the NKG2d antigen is present on the surface of an NK cell. In certain embodiments, the GPRC5d antigen is present on the surface of a tumor cell. Binding of an NKG2d / GPRC5d multispecific antibody to NKG2d present on the surface of an NK cell and GPRC5d present on the surface of a tumor cell can result in, for example, tumor cell killing. In other embodiments, binding of an NKG2d / GPRC5d multispecific antibody to NKG2d present on the surface of an NK cell and GPRC5d present on the surface of a tumor cell can result in, for example, NK cell activation.

[0311] In certain embodiments, the NKp46 antibody comprises a VH region and a VL region. In some embodiments, the NKp46 antibody is a single-chain antibody. In some embodiments, the NKp46 antibody is a single-domain antibody. In some embodiments, the NKp46 antibody is a nanobody. In certain embodiments, the NKp46 antibody is a VHH antibody. In certain embodiments, the NKp46 antibody is a llama antibody. In some embodiments, the NKp46 antibody is not a single-chain antibody. In some embodiments, the NKp46 antibody is not a single-domain antibody. In some embodiments, the NKp46 antibody is not a nanobody. In certain embodiments, the NKp46 antibody is not a VHH antibody. In certain embodiments, the NKp46 antibody is not a llama antibody. In some embodiments, the NKp46 antibody is a multispecific antibody. In other embodiments, the NKp46 is a bispecific antibody. In certain embodiments, the multispecific antibody comprises an antigen-binding fragment of an NKp46 antibody provided herein. In other embodiments, the bispecific antibody comprises an antigen-binding fragment of an NKp46 antibody provided herein.

[0312] In another aspect, provided herein are multispecific antibodies that bind to NKp46. In some embodiments, the multispecific antibody is a bispecific antibody. In some embodiments, the multispecific antibody is a trispecific antibody. In some embodiments, the multispecific antibody is a tetraspecific antibody. In one embodiment, the multispecific NKp46 antibody comprises (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to a second target. In one embodiment, the multispecific NKp46 antibody comprises (a) a first binding domain that binds to NKp46, (b) a second binding domain that binds to a second target, and (c) a third binding domain that binds to a third target. In one embodiment, the multispecific NKp46 antibody comprises (a) a first binding domain that binds to NKp46, (b) a second binding domain that binds to a second target, (c) a third binding domain that binds to a third target, and (d) a fourth binding domain that binds to a fourth target.

[0313] In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to a second target other than NKp46. In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to a second target that binds to an antigen expressed on tumor cells. In some embodiments, the second binding domain binds to BCMA. In some embodiments, the second binding domain binds to GPRC5d.

[0314] In certain embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds to NKp46 having the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds to NKp46 having the VH region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds to NKp46 having the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds to NKp46 having the VH region of any one of the antibodies described herein and the VL region of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds NKp46 having the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described. In some embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds NKp46 having the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein. In some embodiments, provided herein are anti-NKp46 bispecific antibodies comprising a binding domain that binds NKp46 having the VH CDR1, VH CDR2, and VH CDR3 of any one of the antibodies described herein and the VL CDR1, VL CDR2, and VL CDR3 of any one of the antibodies described herein.

[0315] In certain embodiments, the anti-NKp46 antibody is a bispecific antibody. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to BCMA having the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to BCMA having the VH region of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to BCMA having the VL region of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to BCMA having the VH region of the anti-BCMA antibodies provided herein and the VL region of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds BCMA having the VH CDR1, VH CDR2, and VH CDR3 of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds BCMA having the VL CDR1, VL CDR2, and VL CDR3 of the anti-BCMA antibodies provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds BCMA having the VH CDR1, VH CDR2, and VH CDR3 of the anti-BCMA antibodies provided herein and the VL CDR1, VL CDR2, and VL CDR3 of the anti-BCMA antibodies provided herein.

[0316] In certain embodiments, the anti-NKp46 antibody is a bispecific antibody. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH region, VL region, VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and / or VL CDR3 of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH region of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VL region of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH region of the anti-GPRC5d antibody provided herein and the VL region of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH CDR1, VH CDR2, and VH CDR3 of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VL CDR1, VL CDR2, and VL CDR3 of the anti-GPRC5d antibody provided herein. In some embodiments, the anti-NKp46 bispecific antibody further comprises a second binding domain that binds to GPRC5d having the VH CDR1, VH CDR2, and VH CDR3 of an anti-GPRC5d antibody provided herein and the VL CDR1, VL CDR2, and VL CDR3 of an anti-GPRC5d antibody provided herein.

[0317] In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 is as described or derived from an antibody described above. In certain embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 69, 70, and 71, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 72, 73, and 74, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 75, 76, and 77, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 78, 79, and 80, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 81, 82, and 83, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 84, 85, and 86, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 87, 88, and 89, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively.In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 93, 94, and 95, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 96, 97, and 98, respectively. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 67. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 68. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 67, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 68. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH having the amino acid sequence of SEQ ID NO: 67. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VL having the amino acid sequence of SEQ ID NO: 68. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH having the amino acid sequence of SEQ ID NO: 67 and a VL having the amino acid sequence of SEQ ID NO: 68.In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 67. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 68. In some embodiments of the multispecific antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 67, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 68.

[0318] In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the Kabat numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the Chothia numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the AbM numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKp46 are according to the Contact numbering system. In some embodiments, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds to NKp46 are according to the IMGT numbering system.

[0319] In some embodiments of the multispecific NKp46 antibodies provided herein, the first binding domain binds to an NKp46 antigen. In some embodiments, the first binding domain binds to an NKp46 epitope. In some embodiments, the first binding domain specifically binds to NKp46. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an NKp46 antigen. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an NKp46 epitope. In some embodiments, NKp46 is present on the surface of NK cells.

[0320] In another aspect, provided herein is a multispecific antibody that competes for binding to NKp46 with any of the NKp46 antibodies described herein. In another aspect, provided herein is a multispecific antibody that binds to the same epitope as any of the NKp46 antibodies described herein. In another aspect, provided herein is a multispecific NKp46 antibody that binds to an epitope on NKp46 that overlaps with the epitope on NKp46 bound by the NKp46 antibodies described herein.

[0321] In one embodiment, a multispecific antibody is provided that competes with an NKp46 reference antibody for binding to NKp46. In another embodiment, a multispecific NKp46 antibody is provided that binds to the same NKp46 epitope as the NKp46 reference antibody. In another embodiment, a NKp46 antibody is provided that binds to an epitope on NKp46 that overlaps with the epitope on NKp46 bound by the NKp46 reference antibody.

[0322] In one embodiment, the NKp46 reference antibody comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 67, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 68.

[0323] In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the third target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the fourth target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 antigen and the third target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 antigen and the fourth target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the third target is not an NKp46 antigen and the fourth target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 antigen, the third target is not an NKp46 antigen, and the fourth target is not an NKp46 antigen. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 antibodies provided herein, the third target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 antibodies provided herein, the fourth target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 epitope and the third target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 epitope and the fourth target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 antibodies provided herein, the third target is not an NKp46 epitope and the fourth target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is not an NKp46 epitope, the third target is not an NKp46 epitope, and the fourth target is not an NKp46 epitope.In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is BCMA. In some embodiments of the multispecific NKp46 antibodies provided herein, the second target is GPRC5d.

[0324] Binding of a multispecific antibody provided herein to NKp46 present on the surface of an NK cell and binding of a second target antigen present on the surface of a second target cell can, for example, result in killing of the second target cell. In other embodiments, binding of a multispecific antibody provided herein to NKp46 present on the surface of an NK cell and binding of a second target antigen can, for example, result in activation of the NK cell. In some embodiments, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to a cancer antigen present on the surface of a cancer cell. In some embodiments, the antigen on the surface of the cancer cell is a tumor-specific antigen. In some embodiments, the antigen on the surface of the cancer cell is a tumor-associated antigen. In some embodiments, the antigen on the surface of the cancer cell is a neoantigen. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKp46. In some embodiments, NKp46 is present on the surface of an NK cell. In some embodiments, when the bispecific antibody binds to NKp46 on the surface of an NK cell and an antigen on the surface of the cancer cell, the cancer cell is killed.

[0325] Binding of a multispecific antibody provided herein to NKp46 present on the surface of a T cell and binding of a second target antigen present on the surface of a second target cell can, for example, result in killing of the second target cell. In other embodiments, binding of a multispecific antibody provided herein to NKp46 present on the surface of a T cell and binding of a second target antigen can, for example, result in activation of the T cell. In some embodiments, provided herein are bispecific antibodies comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to a cancer antigen present on the surface of a cancer cell. In some embodiments, the antigen on the surface of the cancer cell is a tumor-specific antigen. In some embodiments, the antigen on the surface of the cancer cell is a tumor-associated antigen. In some embodiments, the antigen on the surface of the cancer cell is a neoantigen. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKp46. In some embodiments, NKp46 is present on the surface of a T cell. In some embodiments, when the bispecific antibody binds to NKp46 on the surface of the T cell and an antigen on the surface of the cancer cell, the cancer cell is killed. In some embodiments, the T cell is a gamma delta T cell. In some embodiments, the T cell is a mucosal population of innate lymphoid cells.

[0326] In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to BCMA. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKp46. In some embodiments, NKp46 is present on the surface of an NK cell. In some embodiments, BCMA is present on the surface of a cell. In certain embodiments, NKp46 is present on the surface of an NK cell and BCMA is present on the surface of a cell. In some embodiments, when the bispecific antibody binds to NKp46 on the surface of an NK cell and BCMA on the surface of a cell, the cell bearing BCMA on its surface is killed. In some embodiments, BCMA is present on the surface of a cancer cell. In certain embodiments, NKp46 is present on the surface of an NK cell and BCMA is present on the surface of a cancer cell. In some embodiments, when the bispecific antibody binds to NKp46 on the surface of a cell and BCMA on the surface of a cancer cell, the cancer cell is killed. In certain embodiments, bispecific antibodies are contemplated that comprise any of the NKp46 antibodies provided herein as a first binding domain. In addition, in certain embodiments, bispecific antibodies are also contemplated that comprise any of the NKp46 antibodies provided herein as a first binding domain and a second binding domain that binds to BCMA.

[0327] In some embodiments, the multispecific antibodies provided herein are bispecific antibodies comprising (a) a first binding domain that binds NKp46 and (b) a second binding domain that binds BCMA, wherein the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 101, 102, and 103, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 104, 105, and 106, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 107, 108, and 109, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 110, 111, and 112, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 113, 114, and 115, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 116, 117, and 118, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 119, 120, and 121, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 122, 123, and 124, respectively.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 125, 126, and 127, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 128, 129, and 130, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VL having the amino acid sequence of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99 and a VL having the amino acid sequence of SEQ ID NO: 100.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 99. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 100. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds BCMA comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 99, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 100.

[0328] In another aspect, provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to GPRC5d. In certain embodiments, the first binding domain of the bispecific antibody specifically binds to NKp46. In some embodiments, NKp46 is present on the surface of an NK cell. In some embodiments, GPRC5d is present on the surface of a cell. In certain embodiments, NKp46 is present on the surface of an NK cell and GPRC5d is present on the surface of a cell. In some embodiments, when the bispecific antibody binds to NKp46 on the surface of an NK cell and GPRC5d on the surface of a cell, the cell bearing GPRC5d on its surface is killed. In some embodiments, GPRC5d is present on the surface of a cancer cell. In certain embodiments, NKp46 is present on the surface of an NK cell and GPRC5d is present on the surface of a cancer cell. In some embodiments, when the bispecific antibody binds to NKp46 on the surface of an NK cell and GPRC5d on the surface of a cancer cell, the cancer cell is killed. In certain embodiments, bispecific antibodies are contemplated that include any of the NKp46 antibodies provided herein as a first binding domain. In addition, in certain embodiments, bispecific antibodies are also contemplated that include any of the NKp46 antibodies provided herein as a first binding domain and a second binding domain that binds to GPRC5d.

[0329] In some embodiments, the multispecific antibody provided herein is a bispecific antibody comprising (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to GPRC5d, wherein the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 133, 134, and 135, respectively, and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 136, 137, and 138, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 139, 140, and 141, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 142, 143, and 144, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 145, 146, and 147, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 148, 149, and 150, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of SEQ ID NOs: 151, 152, and 153, respectively; and (ii) a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of SEQ ID NOs: 154, 155, and 156, respectively.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of SEQ ID NOs: 157, 158, and 159, respectively, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of SEQ ID NOs: 160, 161, and 162, respectively. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 131. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises (i) a VH comprising VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 131, and (ii) a VL comprising VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH having the amino acid sequence of SEQ ID NO: 131. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL having the amino acid sequence of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH having the amino acid sequence of SEQ ID NO: 131 and a VL having the amino acid sequence of SEQ ID NO: 132.In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 131. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 132. In some embodiments of the multispecific antibodies provided herein, the second binding domain that binds to GPRC5d comprises a VH comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 131, and a VL comprising an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 132.

[0330] In certain embodiments, multispecific antibodies are provided that comprise the NKp46 antibodies provided herein in a knobs-in-hole format. In certain embodiments, bispecific antibodies are provided that comprise the NKp46 antibodies provided herein in a knobs-in-hole format. In certain embodiments, trispecific antibodies are provided that comprise the NKp46 antibodies provided herein in a knobs-in-hole format. In certain embodiments, tetraspecific antibodies are provided that comprise the NKp46 antibodies provided herein in a knobs-in-hole format. Other specificities can be added to antibodies in a knobs-in-hole format (e.g., adding an scFv to the N- or C-terminus) using methods well known in the art. Furthermore, other formats and methods of making multispecific antibodies are also known in the art and are contemplated. In some embodiments, the NKp46 antibodies provided herein are included in bispecific antibodies. In some embodiments, the NKp46 antibodies provided herein are included in trispecific antibodies. In some embodiments, the NKp46 antibodies provided herein are included in tetraspecific antibodies. In some embodiments, the NKp46 bispecific antibodies provided herein are included in multispecific antibodies.

[0331] In certain embodiments, a multispecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 epitope and a second binding domain that binds to a second epitope, wherein the first NKp46 epitope and the second epitope are not the same. In certain embodiments, a bispecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 epitope and a second binding domain that binds to a second epitope, wherein the first NKp46 epitope and the second epitope are not the same. In certain embodiments, a trispecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 epitope, a second binding domain that binds to a second epitope, and a third binding domain that binds to a third epitope, wherein the first NKp46 epitope, the second epitope, and the third epitope are not the same. In certain embodiments, a tetraspecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 epitope, a second binding domain that binds to a second epitope, a third binding domain that binds to a third epitope, and a fourth binding domain that binds to a fourth epitope, wherein the first NKp46 epitope, the second epitope, the third epitope, and the fourth epitope are not the same. In certain embodiments, a multispecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 antigen and a second binding domain that binds to a second antigen, wherein the first NKp46 antigen and the second antigen are not the same. In certain embodiments, a bispecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 antigen and a second binding domain that binds to a second antigen, wherein the first NKp46 antigen and the second antigen are not the same.In certain embodiments, a triabody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 antigen, a second binding domain that binds to a second antigen, and a third binding domain that binds to a third antigen, wherein the first NKp46 antigen, the second antigen, and the third antigen are not the same. In certain embodiments, a tetrabody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to a first NKp46 antigen, a second binding domain that binds to a second antigen, a third binding domain that binds to a third antigen, and a fourth binding domain that binds to a fourth antigen, wherein the first NKp46 antigen, the second antigen, the third antigen, and the fourth antigen are not the same. In certain embodiments, an NKp46 antibody or antigen-binding fragment thereof provided herein specifically binds to NKp46.

[0332] In some embodiments, the multispecific antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region, and the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKp46 antibody is not a single domain antibody or a nanobody. In some embodiments, the third binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the fourth binding domain comprises a heavy chain variable region and a light chain variable region.

[0333] In certain embodiments, an NKp46 multispecific antibody or antigen-binding fragment thereof binds to a first epitope located on NKp46 and a second epitope on a second target antigen. Provided herein in some embodiments is a multispecific antibody comprising (a) a first binding domain that binds to the NKp46 antigen and (b) a second binding domain that binds to the second target antigen. Provided herein in some embodiments is a multispecific antibody comprising (a) a first binding domain that specifically binds to the NKp46 antigen and (b) a second binding domain that specifically binds to the second target antigen. Provided herein in some embodiments is a multispecific antibody comprising (a) a first binding domain that binds to a first epitope on the NKp46 antigen and (b) a second binding domain that binds to a second epitope on the second target antigen. In some embodiments, provided herein is a multispecific antibody comprising: (a) a first binding domain that specifically binds to a first epitope on the NKp46 antigen; and (b) a second binding domain that specifically binds to a second epitope on a second target antigen.

[0334] In certain embodiments, the NKp46 antigen is on the surface of an NK cell. In certain embodiments, the second target antigen is not NKp46. Binding of an NKp46 multispecific antibody to NKp46 present on the surface of an NK cell and binding of a second target antigen present on the surface of a second target cell can, for example, result in killing of the second target cell. In other embodiments, binding of an NKp46 multispecific antibody to NKp46 present on the surface of an NK cell and binding of a second target antigen can, for example, result in activation of the NK cell.

[0335] In certain embodiments, the NKp46 antigen is present on the surface of a T cell. In certain embodiments, the second target antigen is not NKp46. Binding of an NKp46 multispecific antibody to NKp46 present on the surface of a T cell and binding of a second target antigen present on the surface of a second target cell can result in, for example, killing of the second target cell. In other embodiments, binding of an NKp46 multispecific antibody to NKp46 present on the surface of a T cell and binding of a second target antigen can result in, for example, activation of the T cell. In some embodiments, the T cell is a gamma delta T cell. In some embodiments, the T cell is an innate lymphoid cell.

[0336] In another aspect, provided herein is a multispecific antibody comprising a first binding domain that binds NKp46 and a second binding domain that binds BCMA (a "multispecific NKp46 / BCMA antibody"). In some embodiments, the multispecific NKp46 / BCMA antibody is a bispecific antibody. In some embodiments, the multispecific NKp46 / BCMA antibody is a trispecific antibody. In some embodiments, the multispecific NKp46 / BCMA antibody is a tetraspecific antibody.

[0337] In some embodiments, the multispecific NKp46 / BCMA antibodies provided herein comprise (a) a first binding domain that binds to NKp46 and (b) a second binding domain that binds to BCMA. In one embodiment, the multispecific NKp46 / BCMA antibodies comprise (a) a first binding domain that binds to NKp46, (b) a second binding domain that binds to BCMA, and (c) a third binding domain that binds to a third target. In one embodiment, the multispecific NKp46 / BCMA antibodies comprise (a) a first binding domain that binds to NKp46, (b) a second binding domain that binds to BCMA, (c) a third binding domain that binds to a third target, and (d) a fourth binding domain that binds to a fourth target.

[0338] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH comprising a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of the VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 67. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds to NKp46 comprises a VL comprising a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of the VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 68. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds to NKp46 comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of the VH having the amino acid sequence of SEQ ID NO: 67, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of the VL having the amino acid sequence of SEQ ID NO: 68. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds to NKp46 comprises a VH having the amino acid sequence of SEQ ID NO: 67. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds to NKp46 comprises a VL having the amino acid sequence of SEQ ID NO: 68. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds NKp46 comprises a VH having the amino acid sequence of SEQ ID NO: 67 and a VL having the amino acid sequence of SEQ ID NO: 68.

[0339] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the Kabat numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the Chothia numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the AbM numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the Contact numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain that binds NKp46 are according to the IMGT numbering system.

[0340] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain binds to the NKp46 antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain binds to an NKp46 epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain specifically binds to NKp46. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form the binding site for the NKp46 antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the first binding domain form a binding site for an epitope of NKp46. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, NKp46 is present on the surface of an NK cell.

[0341] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VH that includes a VH CDR1, a VH CDR2, and a VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VL that includes a VL CDR1, a VL CDR2, and a VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises (i) a VH comprising a VH CDR1, VH CDR2, and VH CDR3 having the amino acid sequences of VH CDR1, VH CDR2, and VH CDR3, respectively, of SEQ ID NO: 99, and (ii) a VL comprising a VL CDR1, VL CDR2, and VL CDR3 having the amino acid sequences of VL CDR1, VL CDR2, and VL CDR3, respectively, of SEQ ID NO: 100. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VL having the amino acid sequence of SEQ ID NO: 100. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA comprises a VH having the amino acid sequence of SEQ ID NO: 99 and a VL having the amino acid sequence of SEQ ID NO: 100.

[0342] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the Kabat numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the Chothia numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the AbM numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the Contact numbering system. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the amino acid sequences of the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain that binds BCMA are according to the IMGT numbering system.

[0343] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain binds to a BCMA antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain binds to a BCMA epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain specifically binds to BCMA. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain form a binding site for an antigen of BCMA. In some embodiments, the VH CDR1, VH CDR2, VH CDR3, VL CDR1, VL CDR2, and VL CDR3 of the second binding domain form a binding site for an epitope of BCMA. In some embodiments, BCMA is present on the surface of a tumor cell.

[0344] In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not the NKp46 antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the fourth target is not the NKp46 antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not the NKp46 antigen and the fourth target is not the NKp46 antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not the BCMA antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the fourth target is not the BCMA antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not the BCMA antigen and the fourth target is not a BCMA antigen. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the fourth target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not an NKp46 epitope and the fourth target is not an NKp46 epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not a BCMA epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the fourth target is not a BCMA epitope. In some embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the third target is not a BCMA epitope and the fourth target is not a BCMA epitope.

[0345] In certain embodiments, the target is derived from a mammal. In certain embodiments, the target is derived from a rat. In certain embodiments, the target is derived from a mouse. In certain embodiments, the target is derived from a primate. In certain embodiments, the target is derived from a human.

[0346] In certain embodiments, multispecific NKp46 / BCMA antibodies in a knobs-in-hole format are provided. In certain embodiments, bispecific NKp46 / BCMA antibodies in a knobs-in-hole format are provided. In certain embodiments, trispecific antibodies in a knobs-in-hole format are provided. In certain embodiments, tetraspecific antibodies in a knobs-in-hole format are provided. Other specificities can be added to antibodies in a knobs-in-hole format (e.g., adding an scFv to the N- or C-terminus) using methods well known in the art. Furthermore, other formats and methods of making multispecific antibodies are also known in the art and are contemplated. In some embodiments, the NKp46 / BCMA antibodies provided herein are included in bispecific antibodies. In some embodiments, the NKp46 / BCMA antibodies provided herein are included in trispecific antibodies. In some embodiments, the NKp46 / BCMA antibodies provided herein can be included in tetraspecific antibodies. In some embodiments, the NKp46 / BCMA bispecific antibodies provided herein are included in multispecific antibodies.

[0347] In certain embodiments, the trispecific NKp46 / BCMA antibodies provided herein comprise a first binding domain comprising an NKp46 antibody provided herein that binds to an NKp46 epitope, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA epitope, and a third binding domain that binds to a third epitope, wherein the NKp46 epitope, the BCMA epitope, and the third epitope are not the same. In certain embodiments, a tetraspecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to an NKp46 epitope, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA epitope, a third binding domain that binds to a third epitope, and a fourth binding domain that binds to a fourth epitope, wherein the NKp46 epitope, the BCMA epitope, the third epitope, and the fourth epitope are not the same. In certain embodiments, a trispecific antibody provided herein comprises a first binding domain comprising an NKp46 antibody provided herein that binds to an NKp46 antigen, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA antigen, and a third binding domain that binds to a third antigen, wherein the NKp46 antigen, the BCMA antigen, and the third antigen are not the same. In certain embodiments, the tetraspecific antibodies provided herein bind to an NKp46 antigen, a second binding domain comprising a BCMA antibody provided herein that binds to a BCMA antigen, a third binding domain that binds to a third antigen, a fourth binding domain that binds to a fourth antigen, The NKp46 antigen, the BCMA antigen, the third antigen, and the fourth antigen are not the same. In certain embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds NKp46 specifically binds to NKp46. In other embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the second binding domain that binds BCMA specifically binds to BCMA. In yet other embodiments of the multispecific NKp46 / BCMA antibodies provided herein, the first binding domain that binds NKp46 specifically binds to NKp46 and the second binding domain that binds BCMA specifically binds to BCMA.

[0348] In some embodiments, the multispecific NKp46 / BCMA antibody comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the first binding domain comprises a heavy chain variable region and a light chain variable region, and the second binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the NKp46 antibody is not a single domain antibody or a nanobody. In some embodiments, the third binding domain comprises a heavy chain variable region and a light chain variable region. In some embodiments, the fourth binding domain comprises a heavy chain variable region and a light chain variable region.

[0349] In certain embodiments, an NKp46 / BCMA multispecific antibody or antigen-binding fragment thereof binds to a first epitope located on NKp46 and a second epitope located on BCMA. Provided herein in some embodiments are multispecific NKp46 / BCMA antibodies comprising (a) a first binding domain that binds to the NKp46 antigen and (b) a second binding domain that binds to the BCMA antigen. Provided herein in some embodiments are multispecific NKp46 / BCMA antibodies comprising (a) a first binding domain that specifically binds to the NKp46 antigen and (b) a second binding domain that specifically binds to the BCMA antigen. Provided herein in some embodiments are multispecific NKp46 / BCMA antibodies comprising (a) a first binding domain that binds to a first epitope on the NKp46 antigen and (b) a second binding domain that binds to a second epitope on the BCMA antigen. In some embodiments, provided herein is a multispecific antibody comprising: (a) a first binding domain that specifically binds to a first epitope on the NKp46 antigen; and (b) a second binding domain that specifically binds to a second epitope on the BCMA antigen.

[0350] In certain embodiments, the NKp46 antigen is on the surface of an NK cell. In certain embodiments, the BCMA antigen is on the surface of a tumor cell. Binding of an NKp46 / BCMA multispecific antibody to NKp46 present on the surface of an NK cell and BCMA present on the surface of a tumor cell can result in, for example, tumor cell killing. In other embodiments, binding of an NKp46 / BCMA multispecific antibody to NKp46 present on the surface of an NK cell and BCMA present on the surface of a tumor cell can result in, for example, NK cell activation.

[0351] In certain embodiments, the NKp46 antigen is present on the surface of a T cell. In certain embodiments, the BCMA antigen is present on the surface of a tumor cell. Binding of an NKp46 / BCMA multispecific antibody to NKp46 present on the surface of a T cell and BCMA present on the surface of a tumor cell can result in, for example, tumor cell killing. In other embodiments, binding of an NKp46 / BCMA multispecific antibody to NKp46 present on the surface of a T cell and BCMA present on the surface of a tumor cell can result in, for example, T cell activation. In some embodiments, the T cell is a gamma delta T cell. In some embodiments, the T cell is an innate lymphoid cell.

[0352] In another aspect, provided herein is a multispecific antibody ("multispecific NKp46 / GPRC5d antibody") comprising a first binding domain that binds to NKp46 and...

Claims

1. A multispecific antibody, comprising: (a) a first binding domain that binds to a first antigen expressed on a natural killer (NK) cell; (b) a second binding domain that binds to a second antigen; and A multispecific antibody comprising:

2. The multispecific antibody of claim 1, wherein the first antigen is NKG2-D type II integral membrane protein (NKG2d).

3. The first binding domain comprises: (i) (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 4, a VH CDR2 having the amino acid sequence of SEQ ID NO: 5, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 6; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 10, a VH CDR2 having the amino acid sequence of SEQ ID NO: 11, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 12; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 16, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 18; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 22, a VH CDR2 having the amino acid sequence of SEQ ID NO: 23, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 24; or (e) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 28, a VH CDR2 having the amino acid sequence of SEQ ID NO: 29, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 30; and a heavy chain variable region (VH) comprising: (a) a VL CDR1 having the amino acid sequence of SEQ ID NO:7, a VL CDR2 having the amino acid sequence of SEQ ID NO:8, and a VL CDR3 having the amino acid sequence of SEQ ID NO:9; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 13, a VL CDR2 having the amino acid sequence of SEQ ID NO: 14, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 15; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 19, a VL CDR2 having the amino acid sequence of SEQ ID NO: 20, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 21; (d) a VL CDR1 having the amino acid sequence of SEQ ID NO:25, a VL CDR2 having the amino acid sequence of SEQ ID NO:26, and a VL CDR3 having the amino acid sequence of SEQ ID NO:27; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 31, a VL CDR2 having the amino acid sequence of SEQ ID NO: 32, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 33; and a light chain variable region (VL) comprising: or (ii) (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 36, a VH CDR2 having the amino acid sequence of SEQ ID NO: 37, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 38; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 42, a VH CDR2 having the amino acid sequence of SEQ ID NO: 43, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 44; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 48, a VH CDR2 having the amino acid sequence of SEQ ID NO: 49, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 50; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 54, a VH CDR2 having the amino acid sequence of SEQ ID NO: 55, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 56; or (e) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 60, a VH CDR2 having the amino acid sequence of SEQ ID NO: 61, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 62; and a heavy chain variable region (VH) comprising: (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 39, a VL CDR2 having the amino acid sequence of SEQ ID NO: 40, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 41; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 45, a VL CDR2 having the amino acid sequence of SEQ ID NO: 46, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 47; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO:51, a VL CDR2 having the amino acid sequence of SEQ ID NO:52, and a VL CDR3 having the amino acid sequence of SEQ ID NO:53; (d) a VL CDR1 having the amino acid sequence of SEQ ID NO:57, a VL CDR2 having the amino acid sequence of SEQ ID NO:58, and a VL CDR3 having the amino acid sequence of SEQ ID NO:59; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO:63, a VL CDR2 having the amino acid sequence of SEQ ID NO:64, and a VL CDR3 having the amino acid sequence of SEQ ID NO:65; and a light chain variable region (VL) comprising: The multispecific antibody of claim 2.

4. 4. The multispecific antibody of claim 3, wherein the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 2 and a VL having the amino acid sequence of SEQ ID NO: 3, or wherein the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO:

35.

5. The multispecific antibody of claim 1 , wherein the first antigen is natural killer cell p46-related protein (NKp46).

6. The first binding domain comprises: (i) (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 69, a VH CDR2 having the amino acid sequence of SEQ ID NO: 70, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 71; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 75, a VH CDR2 having the amino acid sequence of SEQ ID NO: 76, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 77; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 81, a VH CDR2 having the amino acid sequence of SEQ ID NO: 82, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 83; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 87, a VH CDR2 having the amino acid sequence of SEQ ID NO: 88, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 89; or (e) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 93, a VH CDR2 having the amino acid sequence of SEQ ID NO: 94, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 95; and a heavy chain variable region (VH) comprising: (ii) (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 72, a VL CDR2 having the amino acid sequence of SEQ ID NO: 73, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 74; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 78, a VL CDR2 having the amino acid sequence of SEQ ID NO: 79, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 80; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 84, a VL CDR2 having the amino acid sequence of SEQ ID NO: 85, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 86; (d) a VL CDR1 having the amino acid sequence of SEQ ID NO: 90, a VL CDR2 having the amino acid sequence of SEQ ID NO: 91, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 92; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 96, a VL CDR2 having the amino acid sequence of SEQ ID NO: 97, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 98; and a light chain variable region (VL) comprising: The multispecific antibody of claim 5 , comprising:

7. The multispecific antibody of claim 6, wherein the first binding domain comprises a VH having the amino acid sequence of SEQ ID NO: 67 and a VL having the amino acid sequence of SEQ ID NO:

68.

8. The method of claim 1, wherein the second antigen is expressed on a tumor cell, and the second antigen is a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA); the second antigen is B cell maturation antigen (BCMA); or The multispecific antibody of any one of claims 1 to 7, wherein the second antigen is G protein-coupled receptor family C group 5 member D (GPRC5d).

9. 2. The multispecific antibody of claim 1, wherein the bispecific antibody is a Bipod scaffold and optionally the first binding domain is a Fab region and the second binding domain is a scFv region.

10. The multispecific antibody induces NK cell-dependent cytotoxicity of the tumor cells in vitro with an IC of less than about 500 picomolar (pM). ５０ 10. The multispecific antibody of claim 8 or 9, wherein the antibody is induced with 50 mM Hsp300 and optionally has an IC50 of less than about 100 pM.

11. Said IC ５０ is assessed using a mixture of NK effector cells and target cells expressing said second antigen, optionally at a ratio of effector cells to target cells selected from the group consisting of about 0.01:1 to about 5:1, about 0.1:1 to about 2:1 and about 1:

1.

12. An antibody that binds to NKG2-D type II integral membrane protein (NKG2d), (i) (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 4, a VH CDR2 having the amino acid sequence of SEQ ID NO: 5, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 6; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 10, a VH CDR2 having the amino acid sequence of SEQ ID NO: 11, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 12; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 16, a VH CDR2 having the amino acid sequence of SEQ ID NO: 17, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 18; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 22, a VH CDR2 having the amino acid sequence of SEQ ID NO: 23, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 24; or (e) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 28, a VH CDR2 having the amino acid sequence of SEQ ID NO: 29, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 30; and a heavy chain variable region (VH) comprising: (a) a VL CDR1 having the amino acid sequence of SEQ ID NO:7, a VL CDR2 having the amino acid sequence of SEQ ID NO:8, and a VL CDR3 having the amino acid sequence of SEQ ID NO:9; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 13, a VL CDR2 having the amino acid sequence of SEQ ID NO: 14, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 15; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 19, a VL CDR2 having the amino acid sequence of SEQ ID NO: 20, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 21; (d) a VL CDR1 having the amino acid sequence of SEQ ID NO:25, a VL CDR2 having the amino acid sequence of SEQ ID NO:26, and a VL CDR3 having the amino acid sequence of SEQ ID NO:27; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 31, a VL CDR2 having the amino acid sequence of SEQ ID NO: 32, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 33; and a light chain variable region (VL) comprising: or (ii) (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 36, a VH CDR2 having the amino acid sequence of SEQ ID NO: 37, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 38; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 42, a VH CDR2 having the amino acid sequence of SEQ ID NO: 43, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 44; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 48, a VH CDR2 having the amino acid sequence of SEQ ID NO: 49, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 50; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 54, a VH CDR2 having the amino acid sequence of SEQ ID NO: 55, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 56; or (e) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 60, a VH CDR2 having the amino acid sequence of SEQ ID NO: 61, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 62; and a heavy chain variable region (VH) comprising: (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 39, a VL CDR2 having the amino acid sequence of SEQ ID NO: 40, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 41; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 45, a VL CDR2 having the amino acid sequence of SEQ ID NO: 46, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 47; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO:51, a VL CDR2 having the amino acid sequence of SEQ ID NO:52, and a VL CDR3 having the amino acid sequence of SEQ ID NO:53; (d) a VL CDR1 having the amino acid sequence of SEQ ID NO:57, a VL CDR2 having the amino acid sequence of SEQ ID NO:58, and a VL CDR3 having the amino acid sequence of SEQ ID NO:59; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO:63, a VL CDR2 having the amino acid sequence of SEQ ID NO:64, and a VL CDR3 having the amino acid sequence of SEQ ID NO:65; and a light chain variable region (VL) comprising: An antibody that binds to NKG2d.

13. An antibody described in claim 12, comprising a VH having the amino acid sequence of SEQ ID NO: 2 and a VL having the amino acid sequence of SEQ ID NO: 3, or a VH having the amino acid sequence of SEQ ID NO: 34 and a VL having the amino acid sequence of SEQ ID NO:

35.

14. An antibody that binds to natural killer cell p46-related protein (NKp46), comprising: (i) (a) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 69, a VH CDR2 having the amino acid sequence of SEQ ID NO: 70, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 71; (b) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 75, a VH CDR2 having the amino acid sequence of SEQ ID NO: 76, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 77; (c) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 81, a VH CDR2 having the amino acid sequence of SEQ ID NO: 82, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 83; (d) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 87, a VH CDR2 having the amino acid sequence of SEQ ID NO: 88, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 89; or (e) a VH complementarity determining region (CDR) 1 having the amino acid sequence of SEQ ID NO: 93, a VH CDR2 having the amino acid sequence of SEQ ID NO: 94, and a VH CDR3 having the amino acid sequence of SEQ ID NO: 95; and a heavy chain variable region (VH) comprising: (ii) (a) a VL CDR1 having the amino acid sequence of SEQ ID NO: 72, a VL CDR2 having the amino acid sequence of SEQ ID NO: 73, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 74; (b) a VL CDR1 having the amino acid sequence of SEQ ID NO: 78, a VL CDR2 having the amino acid sequence of SEQ ID NO: 79, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 80; (c) a VL CDR1 having the amino acid sequence of SEQ ID NO: 84, a VL CDR2 having the amino acid sequence of SEQ ID NO: 85, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 86; (d) a VL CDR1 having the amino acid sequence of SEQ ID NO: 90, a VL CDR2 having the amino acid sequence of SEQ ID NO: 91, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 92; or (e) a VL CDR1 having the amino acid sequence of SEQ ID NO: 96, a VL CDR2 having the amino acid sequence of SEQ ID NO: 97, and a VL CDR3 having the amino acid sequence of SEQ ID NO: 98; and a light chain variable region (VL) comprising: An antibody that binds to NKp46, comprising:

15. The antibody described in claim 14, comprising a VH having the amino acid sequence of SEQ ID NO: 67 and a VL having the amino acid sequence of SEQ ID NO: 68.

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