CLEC2d targeting conjugates and related methods

Abstract

Provided herein are, inter alia, conjugates comprising an targeting agent (e.g., an antibody) that specifically binds to CLEC2D (e.g., hCLEC2D); and at least one CpG oligonucleotide; and methods of manufacturing and pharmaceutical compositions comprising the same. Further provided herein are methods of utilizing the conjugates, including, e.g., methods of treating cancer (e.g., a solid tumor), methods of activating an immune cell (e.g., a CLEC2D expressing immune cell; activating TLR9 expressed by an immune cell; and methods of inhibiting binding of CD161 to CLEC2D expressed on the surface of an immune cell (e.g., a CLEC2D expressing immune cell).

Description

Attorney Docket No.63340.10WO01 CLEC2D TARGETING CONJUGATES AND RELATED METHODS RELATED APPLICATIONS

[0001] This application claims priority to U.S. Serial No.: 63 / 594,232, filed October 30, 2023, the entire contents of which is incorporated herein by reference. 1. FIELD

[0002] This disclosure is related to, inter alia, conjugates comprising a CLEC2D targeting moiety (e.g., an antibody) that specifically binds to CLEC2D (e.g., human CLEC2D (hCLEC2D)); and a CpG oligonucleotide (CpG ODN). The disclosure further relates to pharmaceutical compositions comprising the same; and methods of utilizing the same, including, e.g., methods of treating cancer (e.g., a solid tumor). 2. BACKGROUND

[0003] CLEC2D is a member of the C type lectin receptor superfamily. In humans, the protein is encoded by a single CLEC2D gene that is precent in the human natural killer gene complex. Multiple isoforms of human CLEC2D are known to be produced by alternative splicing, with isoform 1 being the only isoform predominantly expressed at the cell surface. CLEC2D is known to be expressed by e.g., myeloid cells, such as B cells, as well as on activated dendritic cells (e.g., plasmacytoid dendritic cells and classical dendritic cells). Surface CLEC2D is expressed as a disulfide-bonded homodimer and acts as a ligand for type II transmembrane C-type lectin-like receptor CD161. CLEC2D / CD161 interactions play a role in regulating immune responses in various contexts, including, e.g., infectious diseases, autoimmunity, inflammatory conditions and cancer. CLEC2D is further known to, inter alia, bind histones released upon necrotic cell death that e.g., can stimulate inflammation and amplify tissue injury. 3. SUMMARY

[0004] Provided herein are, inter alia, conjugates comprising a targeting agent that specifically binds CLEC2D (e.g., hCLEC2D) (e.g., an anti-CLEC2D antibody) and a CpG ODN; and methods of manufacturing and pharmaceutical compositions comprising the same. Further provided herein are methods of utilizing the conjugates including, e.g., methods of treating cancer (e.g., a solid tumor) and methods of inhibiting CD161 binding to CLEC2D.Attorney Docket No.63340.10WO01

[0005] Accordingly, in one aspect provided herein are conjugates comprising: (a) an antibody that specifically binds to C-type lectin domain family 2 member D (CLEC2D) (e.g., human CLEC2D (hCLEC2D)); and (b) at least one CpG oligodeoxynucleotide (CpG ODN).

[0006] In some embodiments, the antibody comprises or consists of a full-length antibody, a Fab, a Fab', a F(ab')2, a Fab-Fc, a scFv, a scFv-Fc, a (scFv)2-Fc, an Fv, a single domain antibody (sdAb) (e.g., a VHH), a sdAb-Fc (e.g., a VHH-Fc), a (sdAb)2 (e.g., a (VHH)2), or a (sdAb)2-Fc (e.g., a (VHH)2-Fc). In some embodiments, the antibody is a full-length antibody.

[0007] In some embodiments, the antibody is an IgG (e.g., a human IgG (hIgG)) antibody. In some embodiments, the antibody is a hIgG1, hIgG2, hIgG3, or hIgG4 antibody (e.g., a hIgG1 or hIgG4 antibody).

[0008] In some embodiments, the antibody (e.g., full-length antibody) comprises an immunoglobulin (Ig) (e.g., a human Ig (hIg)) Fc region. In some embodiments, the Ig (e.g., hIg) Fc region comprises at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Ig (e.g., hIg) Fc region comprises a hinge region, a CH2 region, and a CH3 region. In some embodiments, the antibody comprises a first Fc region and a second Fc region that associate via at least one covalent (e.g., disulfide) bond. In some embodiments, the Ig is a hIg. In some embodiments, the hIg is a human IgG (hIgG). In some embodiments, the hIgG is hIgG1 or hIgG4.

[0009] In some embodiments, the Ig (e.g., hIg) Fc region comprises one or more amino acid substitutions relative to a reference Ig (e.g., hIg) Fc region that reduces or abolishes one or more of the following effector functions relative to the reference hIg Fc region: antibody dependent cell mediated cytotoxicity (ADCC), complement dependent cytotoxicity (CDC), and / or affinity to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))). In some embodiments, the Ig (e.g., hIg) Fc region does not substantially mediate ADCC, does not substantially mediate CDC, and / or does not bind to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))).

[0010] In some embodiments, the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an amino acid substitution at amino acid position L234, and / or an amino acid substitution at amino acid position L235, numbering according to the EU index of Kabat. In some embodiments, the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanineAttorney Docket No.63340.10WO01 at amino acid position L234 and / or an alanine at amino acid position L235, numbering according to the EU index of Kabat. In some embodiments, the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234, an alanine at amino acid position L235, and / or a glycine, an alanine, or a serine at position P329 numbering according to the EU index of Kabat. In some embodiments, the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234, a serine at amino acid position L235, and / or a glycine, an alanine, or a serine at position P329 numbering according to the EU index of Kabat. In some embodiments, the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position N297, numbering according to the EU index of Kabat.

[0011] In some embodiments, the Ig is hIgG4 and the amino acid sequence of the Fc region comprises an amino acid substitution at amino acid position S228, an amino acid substitution at amino acid position F234, and / or an amino acid substitution at amino acid position L235, numbering according to the EU index of Kabat. In some embodiments, the Ig is hIgG4 and the amino acid sequence of the Fc region comprises a proline at amino acid position S228, an alanine at amino acid position F234, and / or an alanine at amino acid position L235, numbering according to EU index of Kabat. In some embodiments, the Ig is hIgG4 and the amino acid sequence of the Fc region comprises an alanine at amino acid position N297, numbering according to the EU index of Kabat.

[0012] In some embodiments, the antibody comprises a variable heavy chain (VH) region that comprises three complementarity determining regions: VH CDR1, VH CDR2, and VH CDR3; and a variable light chain (VL) region that comprises three complementarity determining regions: VL CDR1, VL CDR2, and VL CDR3.

[0013] In some embodiments, (a) the amino acid sequence of VH CDR1 comprises the amino acid sequence SEQ ID NO: 3, or the amino acid sequence of SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations; (b) the amino acid sequence of VH CDR2 comprises the amino acid sequence SEQ ID NO: 4, or the amino acid sequence of SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations; (c) the amino acid sequence of VH CDR3 comprises the amino acid sequence SEQ ID NO: 5, or the amino acid sequence of SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations; and (d) the amino acid sequence of VL CDR1 comprises the amino acid sequence SEQ ID NO: 6, or the amino acid sequence of SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations; (e) the amino acid sequence of VL CDR2 comprises the amino acid sequence SEQ ID NO: 7, or the aminoAttorney Docket No.63340.10WO01 acid sequence of SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations; and (f) the amino acid sequence of VL CDR3 comprises the amino acid sequence SEQ ID NO: 8, or the amino acid sequence of SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations.

[0014] In some embodiments, the VH region comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 9; and the VL region comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 10.

[0015] In some embodiments, the first moiety comprises a heavy chain (HC) and a light chain (LC), wherein (a) the amino acid sequence of the HC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11; and the amino acid sequence of the LC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 12; or (b) the amino acid sequence of the HC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 13; and the amino acid sequence of the LC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 12.

[0016] In some embodiments, the CpG ODN activates toll-like receptor 9 (TLR9) (i.e., the CpG oligonucleotide is a TLR9 agonist).

[0017] In some embodiments, the CpG ODN is a Class A, Class B, or Class C CpG ODN. In some embodiments, the CpG ODN is a Class C CpG ODN.

[0018] In some embodiments, the CpG ODN comprises a CpG ODN set forth in Table 8.

[0019] In some embodiments, the nucleotide sequence of the CpG ODN comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in Table 8; or comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in Table 8 comprising 1 or more (e.g., 1, 2, 3) but less than 15% (e.g., less than 12%, less than 10%, less than 8%) nucleotide variations (e.g., substitutions, additions, deletions, etc.).

[0020] In some embodiments, the nucleotide sequence of the CpG ODN comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in any one of SEQ ID NOS: 122- 153 (e.g., SEQ ID NO: 122 or 123); or comprises or consists of the nucleotide sequence of anyAttorney Docket No.63340.10WO01 one of the CpG ODNs set forth in any one of SEQ ID NOS: 122-153 (e.g., SEQ ID NO: 122 or 123) and comprising 1 or more (e.g., 1, 2, 3) but less than 15% (e.g., less than 12%, less than 10%, less than 8%) nucleotide variations (e.g., substitutions, additions, deletions, etc.).

[0021] In some embodiments, the nucleotide sequence of the CpG ODN comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in SEQ ID NO: 122; or comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in SEQ ID NO: 122 and comprising 1 or more (e.g., 1, 2, 3) but less than 15% (e.g., less than 12%, less than 10%, less than 8%) nucleotide variations (e.g., substitutions, additions, deletions, etc.).

[0022] In some embodiments, the nucleotide sequence of the CpG ODN comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in SEQ ID NO: 123; or comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in SEQ ID NO: 123 and comprising 1 or more (e.g., 1, 2, 3) but less than 15% (e.g., less than 12%, less than 10%, less than 8%) nucleotide variations (e.g., substitutions, additions, deletions, etc.).

[0023] In some embodiments, the CpG ODN is single stranded.

[0024] In some embodiments, the CpG ODN comprises from about 15-30, 15-25, 15-20, 16- 30, 16-25, 16-20, 17-30, 17-25, 17-20, 18-30, 18-25, 18-20, 19-30, 19-25, or 19-20 nucleotides in length. In some embodiments, the CpG ODN comprises at least about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.

[0025] In some embodiments, the CpG ODN comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more CpG motifs.

[0026] In some embodiments, the CpG ODN comprises one or more phosphodiester, phosphorothioate, phosphorodithioate, methylphosphonate (MP), or methylphosphorothioate internucleoside linkage. In some embodiments, the CpG ODN comprises one or more phosphorothioate, phosphorodithioate, methylphosphonate, or methylphosphorothioate internucleoside linkage. In some embodiments, all of the internucleoside linkages of the CpG ODN are phosphorothioate linkages.

[0027] In some embodiments, the antibody is non-covalently conjugated to the at least one CpG ODN. In some embodiments, the antibody is covalently conjugated to the at least one CpG ODN. In some embodiments, the antibody is directly conjugated to the at least one CpG ODN. In some embodiments, the antibody is indirectly conjugated to the at least one CpG ODN through aAttorney Docket No.63340.10WO01 linker. In some embodiments, the linker is cleavable or non-cleavable. In some embodiments, the conjugate comprises at least 2, 3, 4, 5, 6, or more CpG ODNs. In some embodiments, each of the at least 2, 3, 4, 5, 6, or more CpG ODNs are individually conjugated to the antibody.

[0028] In some embodiments, the conjugate activates a cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell))) upon binding CLEC2D expressed on the surface of the cell and / or internalization of the conjugate into the cell. In some embodiments, upon internalization of the conjugate into an endosome of a cell expressing CLEC2D the conjugate activates toll-like receptor 9 (TLR9) expressed in the endosome. In some embodiments, the conjugate inhibits (or substantially inhibits) binding of (e.g., hCD161) to CLEC2D (e.g., hCLEC2D).

[0029] In one aspect, provided herein are cells comprising a conjugate described herein. In some embodiments, the cell is in vitro, ex vivo, or in vivo. In some embodiments, the cell is in vitro. In some embodiments, the cell is ex vivo. In some embodiments, the cell is in vivo. In some embodiments, the cell is in a human subject.

[0030] In one aspect, provided herein are pharmaceutical compositions comprising a conjugate described herein and a pharmaceutically acceptable excipient.

[0031] In one aspect, provided herein are kits comprising a conjugate described herein or a pharmaceutical composition described herein, and optionally instructions for utilizing any one of the foregoing.

[0032] In one aspect, provided herein are methods of delivering a conjugate or pharmaceutical composition to a cell, the method comprising introducing into a cell a conjugate described herein or a pharmaceutical composition described herein, to thereby deliver the conjugate or pharmaceutical composition into the cell. In some embodiments, the cell is in vitro, ex vivo, or in vivo. In some embodiments, the cell is a subject (e.g., a human subject).

[0033] In one aspect, provided herein are methods of delivering a conjugate, cell, or pharmaceutical composition to a subject, the method comprising administering to the subject a conjugate described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby deliver the conjugate, cell, or pharmaceutical composition to the subject. In some embodiments, the cell is a subject (e.g., a human subject).

[0034] In one aspect, provided herein are methods of activating an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cellAttorney Docket No.63340.10WO01 (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject, the method comprising administering to the subject a conjugate described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby activate an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject. In some embodiments, the cell is a subject (e.g., a human subject).

[0035] In one aspect, provided herein are methods of activating TLR9 in an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject, the method comprising administering to the subject a conjugate described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby activate TLR9 in an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in the subject. In some embodiments, the cell is a subject (e.g., a human subject).

[0036] In one aspect, provided herein are methods of inhibiting binding of CD161 to CLEC2D expressed on the surface of an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject, the method comprising administering to the subject a conjugate described herein, a cell described herein, or a pharmaceutical composition described herein, to thereby activate TLR9 in an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in the subject. In some embodiments, the cell is a subject (e.g., a human subject).

[0037] In one aspect, provided herein are methods of treating a cancer (e.g., solid tumor) in a subject in need thereof, the method comprising administering to the subject a conjugate described herein, a cell described herein, or a pharmaceutical composition described herein, or the pharmaceutical composition of claim 53, to thereby treat the cancer in the subject. In some embodiments, the cell is a subject (e.g., a human subject).

[0038] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a carcinoma. In some embodiments, the cancer is head cancer, neck cancer, head and neck cancer, lung cancer, colon cancer, rectal cancer, colorectal cancer, renal cancer, or ovarian cancer. In some embodiments, the cancer is head and neck squamous cell carcinoma (HNSCC), non-small cell lungAttorney Docket No.63340.10WO01 cancer, colorectal carcinoma, or renal cell carcinoma.

[0039] In one aspect, provided herein are conjugates, cells, and / or pharmaceutical compositions for use in the treatment of a cancer in a subject in need thereof.

[0040] In one aspect, provided herein are conjugates, cells, and / or pharmaceutical compositions for use as a medicament.

[0041] In one aspect, provided herein is the use of a conjugate described herein, a cell described herein, or a pharmaceutical composition described herein for the manufacture of a medicament for the treatment of a cancer in a subject in need thereof. 4. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] FIG.1 is a graphical depiction of the relative level and percent of CLEC2D expressing cells within various tumor infiltrating immune cell subsets, including, T cells, B cells, NK cells, classical dendritic cells (cDCs), plasmacytoid dendritic cells (pDCs), and macrophages, in an ovarian tumor sample.

[0043] FIG. 2 is a bar graph showing the percent of tumor infiltrating CD11B+ myeloid cells expressing CLEC2D in various solid tumors, including head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer (NSCLC), colorectal cancer (CRC), and renal cell carcinoma (RCC).

[0044] FIG. 3 is a line graph showing the normalized binding of an anti-CLEC2D antibody (SEQ ID NOS: 12-13) to CLEC2D expressed on the surface of an engineered cell line.

[0045] FIG. 4 is a line graph showing the normalized binding of CLEC2D to CD161 in the presence of an anti-CLEC2D antibody (SEQ ID NOS: 12-13) or hIgG control.

[0046] FIG. 5 is a bar graph showing the internalization of an anti-CLEC2D antibody (SEQ ID NOS: 12-13) into CLEC2D expressing cells in vitro as compared to an isotype control antibody.

[0047] FIG. 6 is a bar graph showing cellular NFκB activation in each of the indicated treatment groups (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, and medium control).

[0048] FIG.7 is a bar graph showing cellular IRF activation in each of the indicated treatment groups (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, and medium control).Attorney Docket No.63340.10WO01

[0049] FIG. 8 is a histogram showing CLEC2D expression by activated B cells, resting B cells, and isotype control.

[0050] FIG. 9 is a bar graph showing the level of B cell proliferation in in vitro cultured activated B cells treated with the indicated agent (CLEC2B monoclonal antibody control, isotype control, and anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC)).

[0051] FIG. 10 is a bar graph showing the level of CD86+ B cells in in vitro cultured B cells treated with the indicated agent (CLEC2B monoclonal antibody control, isotype control, and anti- CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC)).

[0052] FIG. 11 is a bar graph showing the level of HLA-DR+ B cells in in vitro cultured B cells treated with the indicated agent (CLEC2B monoclonal antibody control, isotype control, and anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC)).

[0053] FIG. 12 is a bar graph showing the level of CD40+ B cells in in vitro cultured B cells treated with the indicated agent (CLEC2B monoclonal antibody control, isotype control, and anti- CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC)).

[0054] FIG. 13 is a histogram showing the expression of CLEC2D by mature plasmacytoid dendritic cells (pDCs).

[0055] FIG. 14 is a bar graph showing the concentration of IFN-α produced by mature plasmacytoid dendritic cells (pDCs) cultured in vitro in the presence of the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, and medium control).

[0056] FIG. 15 is a histogram showing the expression of CLEC2D by inflammatory macrophages and anti-inflammatory (i.e., inhibitory) macrophages.

[0057] FIG. 16 is a series of histograms showing the proliferation of M2 like (inflammatory T cells), M1 like (immune inhibitory T cells), and M2 like (inflammatory T cells) within in vitro cultured tumor associated macrophages treated with anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC).

[0058] FIG. 17 is a bar graph showing the percentage of IFNγ expressing T cells within an in vitro culture of tumor associated macrophages treated with the indicated agent (media control, CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC)).

[0059] FIG.18 is a bar graph showing the percentage of Granzyme B expressing T cells withinAttorney Docket No.63340.10WO01 an in vitro culture of tumor associated macrophages treated with the indicated agent (media control, CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC)).

[0060] FIG. 19 is a bar graph showing the expression of IFNγ from in vitro cultured human peripheral blood mononuclear cells (hPBMCs) treated with the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, CpG-B control, TLR7 / 8 agonist control, and medium control)).

[0061] FIG. 20 is a bar graph showing the expression of IL-6 from in vitro cultured human peripheral blood mononuclear cells (hPBMCs) treated with the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, CpG-B control, TLR7 / 8 agonist control, and medium control)).

[0062] FIG. 21 is a bar graph showing the expression of TNFα from in vitro cultured human peripheral blood mononuclear cells (hPBMCs) treated with the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, CpG-B control, TLR7 / 8 agonist control, and medium control)).

[0063] FIG. 22 is a bar graph showing the expression of IL-1β from in vitro cultured human peripheral blood mononuclear cells (hPBMCs) treated with the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, CpG-B control, TLR7 / 8 agonist control, and medium control)).

[0064] FIG.23 is a bar graph showing the expression of GMCSF from in vitro cultured human peripheral blood mononuclear cells (hPBMCs) treated with the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure as CLEC2D-ISAC), isotype control, CpG ODN control, CpG-B control, TLR7 / 8 agonist control, and medium control)).

[0065] FIG. 24 is a bar graph showing the expression of IL-10 from in vitro cultured human peripheral blood mononuclear cells (hPBMCs) treated with the indicated agent (CLEC2B monoclonal antibody control, anti-CLEC2D-CpG ODN conjugate (referred to in the figure asAttorney Docket No.63340.10WO01 CLEC2D-ISAC), isotype control, CpG ODN control, CpG-B control, TLR7 / 8 agonist control, and medium control)).

[0066] FIG. 25 is a graphical schematic showing an exemplary anti-CLEC2D CpG ODN conjugation method.

[0067] FIG. 26 is a graphical schematic showing an exemplary anti-CLEC2D CpG ODN conjugation and purification method.

[0068] FIG. 27 is a graphical representation of the conjugation and purification workflow to produce the OAR1 & OAR2 species of anti-CLEC2D CpG ODN conjugates.

[0069] FIG.28 is a line graph showing IRF activation in TLR9 expressing THP1 reporter cells post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1 OAR2, Ab-1 OAR1, humanized Ab-26 OAR2, and humanized Ab-26 OAR1) at the indicated concentration.

[0070] FIG. 29A is a bar graph showing the percentage of B cell proliferation post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1 OAR2, Ab-1 OAR1, humanized Ab-26 OAR2, and humanized Ab-26 OAR1) at 400nM. FIG. 29B is a bar graph showing the level of CD80 expression by B cells post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1 OAR2, Ab-1 OAR1, humanized Ab-26 OAR2, and humanized Ab-26 OAR1) at 400nM.

[0071] FIG. 30 is a bar graph showing the level of IFNα expressed by plasmacytoid dendritic cells post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1 OAR2, Ab-1 OAR1, humanized Ab-26 OAR2, and humanized Ab-26 OAR1) at 100nM.

[0072] FIG. 31A is a line graph showing the binding of the indicated anti-CLEC2D antibody (at the indicated concentration) to endogenous CLEC2D expressed on the surface of Nalm6 cells. FIG. 31B is a line graph showing the binding of the indicated anti-CLEC2D antibody (at the indicated concentration) to CLEC2D knockout Nalm6 cells (no binding observed).

[0073] FIG. 32 is a line graph showing the binding of the indicated anti-CLEC2D antibody (at the indicated concentration) to endogenous CLEC2D expressed on the surface of Nalm6 cells.

[0074] FIG. 33A is a line graph showing the binding of the indicated anti-CLEC2D antibody (at the indicated concentration) to endogenous CLEC2D expressed on the surface of Nalm6 cells. FIG. 33B is a line graph showing the binding of the indicated anti-CLEC2D antibody (at the indicated concentration) to CLEC2D knockout Nalm6 cells (no binding observed).

[0075] FIG. 34 is a line graph showing blocking of human CD161 binding to CD161 expressed by HEK cells by the indicated anti-CLEC2D antibody (at the indicated concentration).Attorney Docket No.63340.10WO01

[0076] FIG. 35 is a line graph showing the blocking of signaling mediated through CD161 / CLEC2D binding in Jurkat-CD161 / Raji (CLEC2D+) signaling assay by the indicated anti- CLEC2D antibody (at the indicated concentration).

[0077] FIG. 36 is a bar graph showing the cellular internalization of the indicated anti- CLEC2D antibody (Ratio = MFI (Test Antibody) / MFI (Ab-1)).

[0078] FIG. 37A is a line graph showing the binding of αCLEC2D-CpG conjugates: Ab-1- Odn-1 conjugate, Ab-1-Odn-2 conjugate; and Ab-1 antibody control to endogenous CLEC2D expressed on Raji cells. FIG. 37B is a line graph showing the binding of αCLEC2D-CpG conjugates: Ab-2-Odn-1 conjugate, Ab-2-Odn-2 conjugate; and Ab-2 antibody control to endogenous CLEC2D expressed on Raji cells. FIG. 37C is a line graph showing the binding of αCLEC2D-CpG conjugates: Ab-8-Odn-1 conjugate, Ab-8-Odn-2 conjugate; and Ab-8 antibody control to endogenous CLEC2D expressed on Raji cells.

[0079] FIG.38 is a bar graph showing IRF activation in TLR9 expressing THP1 reporter cells post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1-Odn-1 conjugate, Ab-2-Odn-1 conjugate, and Ab-8-Odn-1 conjugate) (or the indicated control).

[0080] FIG. 39A is a bar graph showing the percentage of B cell proliferation post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1-Odn-1 OAR1 conjugate, Ab-2-Odn-1 conjugate, and Ab-8-Odn-1 conjugate) (or indicated control). FIG.39B is a bar graph showing the level of CD40 expression by B cells post treatment with the indicted αCLEC2D-CpG conjugate (Ab-1-Odn-1 OAR1 conjugate, Ab-2-Odn-1 conjugate, and Ab-8-Odn-1 conjugate) (or indicated control). 5. DETAILED DESCRIPTION

[0081] A “cold” tumor generally refers to a tumor that is poorly infiltrated by T cells and / or NK cells; while a “hot” tumor describes a tumor that is infiltrated by T cells and / or NK cells. Current immunotherapies largely fail to provide clinical benefit for tumors that are poorly infiltrated with immune cells (i.e., cold tumors). The inventors have, inter alia, discovered novel immunostimulatory antibody conjugates (ISACs) that target CLEC2D and deliver CpG ODNs (TLR9 agonists) to CLEC2D expressing immune cells (e.g., myeloid and B cells) that, inter alia, activate the CLEC2D expressing immune cells (e.g., myeloid and B cells), which in turn can stimulate the recruitment of T cells / NK cells, turning a “cold” tumor “hot.” In some embodiments,Attorney Docket No.63340.10WO01 the ISACs can further block CLEC2D binding to CD161, allowing for activation of infiltrating CD161+ T cells and CD161+ NK cells. Without wishing to be bound by theory, in some embodiments, the conjugates described herein function through, inter alia, the binding to CLEC2D expressed on the surface of an immune cell (e.g., myeloid cell, B cell) and subsequent internalization into the cell where the CpG ODN activates TLR9 expressed in the endosome. Signaling from activated TLR9 results in, e.g., the expression of inflammatory cytokines and chemokines, the production of type I interferon, and increased expression of co-stimulatory molecules, which in turn recruits T cells and NK cells to the tumor microenvironment and promotes antigen presentation, T cell activation, and the generation of tertiary lymphoid structures. As such, the conjugates described herein are useful, inter alia, for the treatment of cancer (e.g., solid tumors). As such, the current disclosure provides anti-CLEC2D-CpG ODN conjugates and their use in, inter alia, pharmaceutical compositions, and methods of treating diseases (e.g., cancer (e.g., solid tumors)). TABLE OF CONTENTS 5.1 Definitions 5.2 Conjugates 5.3 CLEC2D Targeting Agents 5.3.1 Exemplary Anti-CLEC2D Antibodies 5.3.2 Exemplary Properties of CLEC2D Binding Proteins 5.3.2.1 Affinity of CLEC2D Targeting Moiety 5.3.2.2 Inhibition of CD161 Binding to CLEC2D 5.3.3 Ig Constant Regions 5.3.3.1 Ig Effector Function 5.3.3.2 Promotion of Heterodimerization 5.3.3.3 Ig Constant Region Variations and Manipulations for Site Specific Conjugation 5.3.4 Methods of Making Proteins (e.g., Targeting Agents) 5.4 CpG ODNs 5.4.1 Overall Length 5.4.2 CpG Motif Content 5.4.3 Internucleoside Linkage ModificationsAttorney Docket No.63340.10WO01 5.4.4 Methods of Making CpG ONDs 5.5 Ratio of Targeting Agent to Molecular Payload 5.6 Linkers 5.6.1 Cleavable Linkers 5.6.2 Non-Cleavable Linkers 5.6.3 Linker Conjugation 5.7 Conjugation 5.7.1 Exemplary Anti-CLEC2D Antibody – CpG ODN Conjugate 5.8 Cells 5.9 Pharmaceutical Compositions 5.10 Methods of Use 5.10.1 Methods of Delivery 5.10.2 Methods of Activating an Immune Cell 5.10.3 Methods of Activating TLR9 5.10.4 Methods of Inhibiting Binding of CD161 to CLEC2D 5.10.5 Methods of Treating Cancer (e.g., Solid Tumors) 5.11 Kits 5.1 Definitions

[0082] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0083] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed.

[0084] In this disclosure, the use of the singular includes the plural unless specifically stated otherwise. For example, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.Attorney Docket No.63340.10WO01

[0085] It is understood that wherever aspects are described herein with the language “comprising,” otherwise analogous aspects described in terms of “consisting of” and “consisting essentially of” are also provided herein.

[0086] The term “and / or” where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or” as used in a phrase such as “A and / or B” herein is intended to include “A and B,” “A or B,” “A” (alone), and “B” (alone). Likewise, the term “and / or” as used in a phrase such as “A, B, and / or C” is intended to encompass each of the following aspects: 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).

[0087] As described herein, any concentration range, percentage range, ratio range or integer range is to be understood to include the value of any integer within the recited range and, when appropriate, fractions thereof (such as one tenth and one hundredth of an integer), unless otherwise indicated.

[0088] The term “about” refers to a value or composition that is within an acceptable error range for the particular value or composition as determined by one of ordinary skill in the art, which will depend in part on how the value or composition is measured or determined, i.e., the limitations of the measurement system. When particular values or compositions are provided in the disclosure, unless otherwise stated, the meaning of “about” should be assumed to be within an acceptable error range for that particular value or composition.

[0089] Where proteins are described herein, it is understood that nucleic acid molecules (e.g., RNA (e.g., mRNA) or DNA nucleic acid molecules) encoding the protein are also provided herein.

[0090] Where proteins, nucleic acid molecules, vectors, carriers, etc. are described herein, it is understood that isolated forms of the proteins, nucleic acid molecules, vectors, carriers, etc. are also provided herein.

[0091] Where proteins, nucleic acid molecules, etc. are described herein, it is understood that recombinant forms of the proteins, nucleic acid molecules, etc. are also provided herein.

[0092] Where polypeptides or sets of polypeptides are described herein, it is understood that proteins comprising the polypeptides or sets of polypeptides folded into their three-dimensional structure (i.e., tertiary or quaternary structure) are also provided herein and vice versa.

[0093] Where proteins are described herein, it is understood that polypeptides comprising the same amino acid sequence either linear or folded into their three-dimensional structure (i.e.,Attorney Docket No.63340.10WO01 tertiary or quaternary structure) are also provided herein.

[0094] As used herein, the term “administering” refers to the physical introduction of an agent (e.g., a conjugate described herein), e.g., a therapeutic agent (or a precursor of an agent (e.g., a precursor of a therapeutic agent) that is metabolized or altered within the body of the subject to produce the agent (e.g., therapeutic agent) in vivo) to a subject, using any of the various methods and delivery systems known to those skilled in the art. Administering can also be performed, for example, once, a plurality of times, and / or over one or more extended periods. The term administering includes self-administration by a subject to the subject and administration to the subject by a third party.

[0095] The terms “agent” and “moiety” are used interchangeably herein and are used generically to describe any macro or micro molecule (and any combination thereof). Exemplary agents include, but are not limited proteins, peptides, nucleic acid molecules (e.g., DNA, RNA), small molecules, carbohydrates, lipids, synthetic polymers (e.g., polymers of PEG), conjugates (e.g., described herein), and any combination of the foregoing. Agents may contain more than one individual agent (wherein the individual agents are the same or different). For example, an agent may comprise an antibody (e.g., a targeting agent described herein (e.g., an anti-CLEC2D antibody)) and an oligonucleotide (e.g., a CpG ODN (e.g., a CpG ODN described herein)). Agents as defined herein include e.g., conjugates described herein.

[0096] As used herein, the term “affinity” refers to the strength of the binding of one protein (e.g., an Antibody) to another protein (e.g., an Antigen). The affinity of a protein is measured by the dissociation constant Kd, defined as [Antibody] x [Antigen] / [Antibody-Antigen] where [Antibody-Antigen] is the molar concentration of the Antibody-Antigen complex, [Antibody] is the molar concentration of the unbound Antibody and [Ligand] is the molar concentration of the unbound Antigen. The affinity constant Ka is defined by 1 / Kd. Standard methods of measuring affinity are known to the person of ordinary skill in the art. Exemplary methods of measuring affinity include, surface plasmon resonance (SPR) (e.g., BIAcore®-based assay), a common method known in the art (see, e.g., Wilson, Science 295:2103, 2002; Wolff et al., Cancer Res. 55:2560, 1993; and U.S. Patent Nos. 5,283,173, 5,468,614, the full contents of each of which are incorporated by reference herein for all purposes).

[0097] As used herein, the term “antibody” or “antibodies” is used in the broadest sense and encompasses various immunoglobulin (Ig) (e.g., human Ig (hIg), murine Ig (mIg)) structures,Attorney Docket No.63340.10WO01 including, but not limited to monoclonal antibodies, polyclonal antibodies, multispecific (e.g., bispecific, trispecific) antibodies, and antibody fragments so long as they exhibit the desired antigen-binding activity (i.e., antigen binding fragments or variants). The term antibody thus includes, for example, full-length antibodies; antigen-binding fragments of full-length antibodies; molecules comprising antibody CDRs, VH regions, and / or VL regions; and antibody-like scaffolds (e.g., fibronectins). Examples of antibodies include, without limitation, monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies, human antibodies, humanized antibodies, chimeric antibodies, camelized antibodies, intrabodies, a variable domain of a new antigen receptor beta-lactamase (VNAR fragments), affybodies, diabodies, tribodies, heteroconjugate antibodies, antibody-drug conjugates, single domain antibodies (e.g.,VHH, (VHH)2), single chain antibodies, single-chain Fvs (scFv; (scFv)2), Fab fragments (e.g., Fab, single chain Fab (scFab), F(ab’)2 fragments, disulfide-linked Fvs (sdFv), Fc fusions (e.g., Fab-Fc, scFv- Fc, VHH-Fc, (scFv)2-Fc, (VHH)2-Fc), and antigen-binding fragments of any of the above, and conjugates or fusion proteins comprising any of the above. Antibodies can be of Ig isotype (e.g., IgG, IgE, IgM, IgD, or IgA), any class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 or IgA2), or any subclass (e.g., IgG2a or IgG2b) of Ig). In certain embodiments, antibodies described herein are IgG antibodies, or a class (e.g., human IgG1or IgG4) or subclass thereof. In certain embodiments, antibodies described herein are mIgG antibodies, or a class (e.g., mIgG1 or mIgG2a) or subclass thereof. In some embodiments, the antibody is a human, humanized, or chimeric IgG1 or IgG4 monoclonal antibody. In some embodiments, the term antibodies refers to a monoclonal or polyclonal antibody population. The term antibody also includes “antibody-like scaffolds” are defined herein. Antibodies described herein can be produced by any standard methods known in the art, e.g., recombinant production in host cells, see, e.g., § 5.3.4; or synthetic production.

[0098] As used herein, the term “antibody-like scaffold” refers to non-Ig based antigen binding domain. Various antibody-like scaffolds are known in the art. For example, 10th type III domain of fibronectin (e.g., AdNectins®) and designed ankyrin repeat proteins (e.g., DARPins®) have been used as alternative scaffolds for antigen-binding domains, see, e.g., Gebauer and Skerra, Engineered protein scaffolds as next-generation antibody therapeutics. Curr Opin Chem Biol 13:245-255 (2009) and Stumpp et al., Darpins: A new generation of protein therapeutics. Drug Discovery Today 13: 695-701 (2008), the full contents of each of which is incorporated by reference herein for all purposes. Exemplary antibody-like scaffolds include, but are not limitedAttorney Docket No.63340.10WO01 to, lipocalins (see, e.g., US7250297) (e.g., Anticalin®), protein A-derived molecules such as z- domains of protein a (see, e.g., US5831012) (e.g., Affibody®), A domains of membrane receptors stabilized by disulfide bonds and Ca2+ (see, e.g., US7803907) (e.g., Avimer / Maxibody®), a serum transferrin (see, e.g., US2004023334) (e.g., Transbody®); a designed ankyrin repeat protein (see, e.g., US7417130) (e.g., DARPin®), a fibronectin (see, e.g., US6818418) (e.g., AdNectin®), a C- type lectin domain (see, e.g., US2004132094) (e.g., Tetranectin®); a human gamma-crystallin or ubiquitin (see, e.g., US7838629) (e.g., Affilin®); a kunitz type domain of human protease inhibitors (see, e.g., US2004209243), C-Type Lectins (see, e.g., US2004132094) (e.g., Tetranectins®), cysteine knots or knottins (see, e.g., US7186524) (e.g., Microbodies®), nucleic acid aptamers (see, e.g., US5475096), thioredoxin A scaffold (see, e.g., US6004746) (peptide aptamers), and 10th type III domain of fibronectin (see, e.g., US6818418) (e.g., AdNectins®), and cystine-dense peptides (see, e.g., WO2023023031). Additional exemplary antibody-like scaffolds are known in the art and for example described in Storz U. Intellectual property protection: strategies for antibody inventions. MAbs. 2011;3(3):310-317. doi:10.4161 / mabs.3.3.15530. The entire contents of each of the foregoing references is incorporated herein by reference for all purposes. Antibody like scaffolds include e.g., naturally occurring antigen binders, variant (e.g., functional variants) of naturally occurring antigen binders, fragments (e.g., functional fragments) of naturally occurring antigen binders, and synthetic antigen binders (i.e., not naturally occurring antigen binders).

[0099] As used herein, the term “antibody dependent cell mediated cytotoxicity” or “ADCC” refers to an immune mechanism leading to the lysis of antibody (or an Fc region containing protein) (e.g., an Ig Fc containing fusion protein)-coated target cells by immune effector cells (e.g., NK cells). As used herein, the term “reduced ADCC” and the like refers to either a reduction in the number of target cells that are lysed in a given time, at a given concentration of antibody (or an Ig Fc region containing protein) (e.g., an Fc region containing fusion protein) in the medium surrounding the target cells, by the mechanism of ADCC defined above, and / or an increase in the concentration of antibody (or an Fc region containing protein) in the medium surrounding the target cells, required to achieve the lysis of a given number of target cells in a given time, by the mechanism of ADCC defined above. The reduction in ADCC is relative to the ADCC mediated by the same antibody (or an Fc region containing protein) produced by the same type of host cells, using the same standard production, purification, formulation and storage methods (which areAttorney Docket No.63340.10WO01 known to those skilled in the art), but that has not been engineered (e.g., does not comprise one or more amino acid variation, e.g., amino acid substitution, that mediates a decrease in ADCC). For example the reduction in ADCC mediated by an antibody (or an Fc region containing protein) comprising in its Fc region an amino acid substitution that reduces ADCC, is relative to the ADCC mediated by the same antibody (or an Fc region containing protein) without said amino acid substitution in the Fc region.

[0100] The terms “cancer” and “tumor” are used interchangeably herein and refer to a broad group of various diseases characterized by the uncontrolled growth of abnormal cells in the body. Unregulated cell division and growth results in the formation of malignant tumors that can invade neighboring tissues and may also metastasize to distant parts of the body through, e.g., the lymphatic system or bloodstream.

[0101] As used herein, the term “CD161” refers to the type II transmembrane C-type lectin- like receptor expressed, e.g., by natural killer cells. CD161 is also commonly referred to in the art as Killer cell lectin-like receptor subfamily B member 1 (KLRB1). The amino acid sequence of a reference human CD161 (hCD161) protein is set forth in SEQ ID NO: 2 (UniProt Ref.: Q12918- 1).

[0102] As used herein, the term “CDR” or “complementarity determining region” refers to the noncontiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. These particular regions have been described by Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991), the entire contents of each of which is incorporated herein by reference for all purposes. Unless otherwise specified, the term “CDR” is a CDR as defined by Kabat et al., J. Biol. Chem. 252, 6609-6616 (1977) and Kabat et al., Sequences of protein of immunological interest. (1991). A person of ordinary skill in the art would be able to determine the CDRs as defined by another scheme, e.g., Chothia, IMGT, using ordinary methods known in the art.

[0103] The terms “CH1” and “CH1 region” are used interchangeably herein and refer to the first constant region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference hIgG1 CH1 region is set forth in SEQ ID NO: 30; and the amino acid sequence of an exemplary reference hIgG4 CH1 region is set forth in SEQ ID NO: 43.

[0104] The terms “CH2” and “CH2 region” are used interchangeably herein and refer to the second constant region of an immunoglobulin heavy chain. The amino acid sequence of anAttorney Docket No.63340.10WO01 exemplary reference hIgG1 CH2 region is set forth in SEQ ID NO: 32; and the amino acid sequence of an exemplary reference hIgG4 CH2 region is set forth in SEQ ID NO: 45.

[0105] The terms “CH3” and “CH3 region” are used interchangeably herein and refer to the third constant region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference hIgG1 CH3 region is set forth in SEQ ID NO: 33; and the amino acid sequence of an exemplary reference hIgG4 CH3 region is set forth in SEQ ID NO: 46.

[0106] As used herein, the term “Class A” in reference to a CpG ODN refers to a CpG ODN comprising a natural phosphodiester (PO) backbone central CpG-containing palindromic motif and a phosphorothioated (PS) modified 3' poly-G string.

[0107] As used herein, the term “Class B” in reference to a CpG ODN refers to a CpG ODN comprising a full phosphorothioated (PS) modified backbone.

[0108] As used herein, the term “Class C” in reference to a CpG ODN refers to a CpG ODN comprising a full phosphorothioated (PS) modified backbone and a CpG-containing palindromic motif.

[0109] As used herein, the term “CLEC2D” or “C-type lectin domain family 2 member D” refers to the C type lectin receptor that, inter alia, has been reported to be capable of binding histones released upon necrotic cell death. The amino acid sequence of a reference human CLEC2D (hCLEC2D) isoform is set forth in SEQ ID NO: 1 (UniProt Ref.: Q9UHP7-1). Multiple isoforms of hCLEC2D are known produced by alternative splicing.

[0110] As used herein, the term “conjugation” refers to the operable connection (e.g., chemical conjugation) of at least a first agent (e.g., a CpG ODN (e.g., a CpG ODN described herein)) with a second agent (e.g., a targeting agent (e.g., a targeting agent described herein (e.g., an anti- CLEC2D antibody))). The first agent can be directly connected to the second agent or indirectly connected (e.g., through a linker (e.g., as described herein)). Methods of operably connected two agents (e.g., chemical conjugation methods) are well known in the art, as are commercially available conjugation reagents and kits, with detailed instructions for their use readily available from the commercial suppliers. Operable connection (e.g., chemical conjugation) includes both covalent and non-covalent conjugation. In some embodiments, the operable connection (e.g., chemical conjugation) comprises the covalent linkage of the first agent (e.g., a CpG ODN (e.g., a CpG ODN described herein)) with the second agent (e.g., a targeting agent (e.g., a targeting agent described herein (e.g., an anti-CLEC2D antibody))).Attorney Docket No.63340.10WO01

[0111] The terms “constant region” and “constant domain” are used interchangeably herein and refer to a carboxyl terminal portion of a light and / or heavy chain of a full-length antibody which is not directly involved in binding of an antibody to antigen, but which can exhibit various effector functions, such as interaction with an Ig Fc receptor (e.g., Fc gamma receptor). The constant region of an Ig molecule generally has a more conserved amino acid sequence relative to an Ig variable domain.

[0112] As used herein the term “CpG motif” refers to an unmethylated cytosine-phosphate- guanine (CpG) dinucleotide (i.e., a cytosine (C) followed by a guanine (G)) linked by a phosphate bond or a phosphodiester backbone or other internucleoside linkage, such as phosphorothioate, phosphorodithioate, methylphosphonate, or methylphosphorothioate. Phosphorothioate, phosphorodithioate, methylphosphonate and methylphosphorothioate are stabilizing internucleotide linkages, while phosphodiester is a naturally occurring internucleoside linkage.

[0113] As used herein, the term “CpG oligodeoxynucleotide” or “CpG ODN” refers to an oligonucleotide (e.g., DNA oligonucleotide) that comprises at least one CpG motif. Generally, CpG ODNs are capable of activating TLR9 (e.g., expressed by a cell). Any suitable CpG oligonucleotide known to those skilled in the art can be used in the invention in view of the present disclosure.

[0114] The terms “DNA” and “polydeoxyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple deoxyribonucleotides that are polymerized via phosphodiester bonds. Deoxyribonucleotides are nucleotides in which the sugar is deoxyribose.

[0115] The term “effector function” when used in reference to an Ig Fc region or a protein comprising an Ig Fc region (e.g., a full-length antibody) refers to those biological activities attributable to the Ig Fc region of a typical full-length antibody, which therefore vary with the antibody isotype. Antibody effector functions include, but are not limited to, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement dependent cytotoxicity (CDC), Fc receptor binding (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa)), and Clq binding.

[0116] As used herein, the term “EU numbering system” refers to the EU numbering convention for the constant regions of an antibody, as described in Edelman, G.M. et al., Proc. Natl. Acad. USA, 63, 78-85 (1969) and Kabat et al, Sequences of Proteins of Immunological Interest, U.S. Dept. Health and Human Services, 5th edition, 1991, the entire contents of each ofAttorney Docket No.63340.10WO01 which is incorporated herein by reference for all purposes.

[0117] As used herein, the term “Fab” refers to an antigen binding domain that comprises a Fab heavy chain that comprises from N- to C-terminus a VH region and a CH1 region; and a light chain comprising from N- to C-terminus a VL region and a CL region; and wherein the Fab heavy chain and the light chain associate to form an antigen binding domain.

[0118] The term “Fab-Fc” as used herein refers to an antibody that comprises a Fab operably linked to an Fc region.

[0119] As used herein, the term “Fc region” refers to the C-terminal region of a Ig (e.g., a human Ig) heavy chain that comprises from N- to C-terminus at least a CH2 region operably connected to a CH3 region. In some embodiments, the Fc region comprises an Ig hinge region or at least a portion of an Ig hinge region operably connected to the N-terminus of the CH2 region. In some embodiments, the Fc region is engineered relative to a reference Fc region (e.g., comprises one or more amino acid modification), see, e.g., § 5.3.3.1, 5.3.3.2, 5.3.3.3. Additional examples of proteins with engineered Fc regions can be found in Saunders 2019 (K. O. Saunders, “Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life,” 2019, Frontiers in Immunology, V. 10, Art. 1296, pp. 1-20, the entire contents of which is incorporated herein by reference for all purposes).

[0120] As used herein, the terms “first” and “second” with respect to Fc regions etc., are used for convenience of distinguishing when there is more than one of each type of moiety. Use of these terms is not intended to confer a specific order or orientation in the protein unless explicitly so stated. For example, an antibody described herein (e.g., in the case of a full-length antibody) may contain two Fc regions that associate e.g., via one or more covalent (e.g., disulfide) bond.

[0121] As used herein, the term “framework region” or “FR region” refers to the amino acid residues that are part of the variable region of an antibody, but are not part of the CDRs (e.g., using the Kabat definition of CDRs).

[0122] As used herein, the term “full-length antibody” refers to an antibody having a structure substantially similar to a native antibody structure (i) a first Ig light chain comprising from N- to C-terminus a light chain variable region (VL) region and a light chain constant region (CL) region; (ii) a first Ig heavy chain comprising from N- to C-terminus a heavy chain variable region (VH) region, a CH1 region, a hinge region, a CH2 region, and a CH3 region; (iii) a second Ig heavy chain comprising from N- to C-terminus a VH region, a CH1 region, a hinge region, a CH2 region,Attorney Docket No.63340.10WO01 and a CH3 region; (iv) a second Ig light chain comprising from N- to C-terminus a VL region and a VH region; wherein said first light chain and said first heavy chain associate to form a first antigen binding domain; wherein said second light chain and said second heavy chain associate to form a second antigen binding domain; and wherein said first heavy chain and said second heavy chain associate to form a dimer. In some embodiments, the two heavy chains comprise a substantially identical amino acid sequence; and the two light chains comprise a substantially identical amino acid sequence. In some embodiments, the two heavy chains comprise a substantially identical amino acid sequence except for one or more amino acid modifications that promote heterodimerization of the correct heavy chains (e.g., as described herein); and the two light chains comprise a substantially identical amino acid sequence. Antibody chains may be substantially identical but not entirely identical if they differ due to post-translational modifications, such as C-terminal cleavage of lysine residues, alternative glycosylation patterns, etc.

[0123] The term “functional variant” as used herein in reference to a protein refers to a protein that comprises at least one but no more than 20%, not more than 15%, not more than 12%, no more than 10%, no more than 8% amino acid variation (e.g., substitution, deletion, addition) compared to the amino acid sequence of a reference protein, wherein the protein retains at least one particular function of the reference protein. Not all functions of the reference protein (e.g., wild type) need be retained by the functional variant of the protein. In some instances, one or more functions are selectively reduced or eliminated. In some embodiments, the reference protein is a wild type protein.

[0124] The term “functional fragment” as used herein in reference to a protein refers to a fragment of a reference protein that retains at least one particular function. Not all functions of the reference polypeptide or protein need be retained by a functional fragment of the protein. In some instances, one or more functions are selectively reduced or eliminated. In some embodiments, the reference protein is a wild type protein.

[0125] As used herein, the term “heavy chain” refers to the portion of an immunoglobulin (e.g., a human Ig) that typically comprises from N- to C-terminus a heavy chain variable region (VH), a CH1 region, a hinge region, a CH2 region, and a CH3 region. The constant regions of the heavy chain (i.e., the CH1 region, the hinge region, the CH2 region, and the CH3 region) can be any distinct isotype, for example, human alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (µ),Attorney Docket No.63340.10WO01 based on the amino acid sequence of the constant domain, which give rise to the hIgA, hIgD, IgE, hIgG, and hIgM classes of human antibodies, respectively, including subclasses of hIgG, e.g., hIgG1, hIgG2, hIgG3, and hIgG4. As used herein, the term “heavy chain” when used in reference to a human antibody can refer to any distinct type, e.g., alpha (α), delta (δ), epsilon (ε), gamma (γ), and mu (µ), based on the amino acid sequence of the constant domain, which give rise to human IgA, IgD, IgE, IgG, and IgM classes of antibodies, respectively, including subclasses of human IgG, e.g., IgG1, IgG2, IgG3, and IgG4.

[0126] The terms “hinge” or “hinge region” are used interchangeably herein and refer to the hinge region of an immunoglobulin heavy chain. The amino acid sequence of an exemplary reference hIgG1 hinge region is set forth in SEQ ID NO: 31; and the amino acid sequence of an exemplary reference hIgG4 hinge region is set forth in SEQ ID NO: 44.

[0127] As used herein, the term “isolated” with reference to an agent (e.g., a protein, nucleic acid molecule, etc.) refers to an agent (e.g., a protein, nucleic acid molecule, etc.) that is substantially free of other cellular components with which it is associated in the natural state.

[0128] As used herein, the term “modified nucleotide,” “nucleotide modification,” or use of the term “modification” and the like in reference to a nucleotide or nucleic acid sequence refers to a nucleotide comprising a chemical modification, e.g., a modified sugar moiety, a modified nucleobase, and / or a modified internucleoside linkage, or any combination thereof. In certain embodiments of the instant disclosure, inclusion of a deoxynucleotide - which is acknowledged as a naturally occurring form of nucleotide - if present within an RNA oligonucleotide is considered to constitute a modified nucleotide.

[0129] The terms “nucleic acid molecule,” “polynucleotide,” and “oligonucleotide” are used interchangeably herein and refer to a polymer of DNA or RNA. The nucleic acid molecule can be single-stranded or double-stranded; contain natural, non-natural, or altered nucleotides; and contain a natural, non-natural, or altered internucleoside linkage, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule. Nucleic acid molecules include, but are not limited to, all nucleic acid molecules which are obtained by any means available in the art, including, without limitation, recombinant means, e.g., the cloning of nucleic acid molecules from a recombinant library or a cell genome, using ordinary cloning technology and polymerase chain reaction, and the like, and by synthetic means. The skilled artisan will appreciate that, except where otherwiseAttorney Docket No.63340.10WO01 noted, nucleic acid sequences set forth in the instant application will recite thymidine (T) in a representative DNA sequence but where the sequence represents RNA (e.g., mRNA), the thymidines (Ts) would be substituted for uracils (Us). Thus, any of the RNA polynucleotides encoded by a DNA identified by a particular sequence identification number may also comprise the corresponding RNA (e.g., mRNA) sequence encoded by the DNA, where each thymidine (T) of the DNA sequence is substituted with uracil (U).

[0130] As used herein, the term “operably connected” refers to the linkage of two agents in a functional relationship. For example, a polypeptide is operably connected to another polypeptide when they are linked (either directly or indirectly via a peptide linker) in frame such that both polypeptides are functional. Or for example, a transcription regulatory polynucleotide e.g., a promoter, enhancer, or other expression control element is operably linked to a polynucleotide that encodes a protein if it affects the transcription of the polynucleotide that encodes the protein. The term “operably connected” also refers for example to the conjugation of a first agent (e.g., an antibody (e.g., an anti-CLEC2D antibody))) to a second agent (e.g., an oligonucleotide (e.g., a CpG ODN)) wherein the first and second agent are both capable of mediating their function.

[0131] The determination of “percent identity” between two sequences (e.g., protein (amino acid sequences) or oligonucleotide (nucleic acid sequences)) can be accomplished using a mathematical algorithm. Determinations of identity (as described herein) are independent of nucleotide chemical modifications (e.g., as described herein). For example, (mC) is identical to (C) for the purposes of determining identity. A specific, non-limiting example of a mathematical algorithm utilized for the comparison of two sequences is the algorithm of Karlin S & Altschul SF (1990) PNAS 87: 2264-2268, modified as in Karlin S & Altschul SF (1993) PNAS 90: 5873-5877, each of which is herein incorporated by reference in its entirety. Such an algorithm is incorporated into the NBLAST and XBLAST programs of Altschul SF et al., (1990) J Mol Biol 215: 403, which is herein incorporated by reference in its entirety. BLAST nucleotide searches can be performed with the NBLAST nucleotide program parameters set, e.g., for score=100, wordlength=12 to obtain nucleotide sequences homologous to a nucleic acid molecule described herein. BLAST protein searches can be performed with the XBLAST program parameters set, e.g., to score 50, wordlength=3 to obtain amino acid sequences homologous to a protein molecule described herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul SF et al., (1997) Nuc Acids Res 25: 3389-3402, which is herein incorporatedAttorney Docket No.63340.10WO01 by reference in its entirety. Alternatively, PSI BLAST can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI Blast programs, the default parameters of the respective programs (e.g., of XBLAST and NBLAST) can be used (see, e.g., National Center for Biotechnology Information (NCBI) on the worldwide web, ncbi.nlm.nih.gov). Another specific, non-limiting example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, 1988, CABIOS 4:11-17, which is herein incorporated by reference in its entirety. Such an algorithm is incorporated in the ALIGN program (version 2.0) which is part of the GCG sequence alignment software package. When utilizing the ALIGN program for comparing 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. In calculating percent identity, typically only exact matches are counted.

[0132] As used herein, the term “pharmaceutical composition” means a composition that is suitable for administration to an animal, e.g., a human subject, and comprises a therapeutic agent (e.g., a conjugate described herein) and a pharmaceutically acceptable carrier or diluent. A “pharmaceutically acceptable carrier or diluent” means a substance intended for use in contact with the tissues of human beings and / or non-human animals, and without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable therapeutic benefit / risk ratio.

[0133] As used herein, the term “plurality” means 2 or more (e.g., 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 9 or more, or 10 or more).

[0134] As used herein, the terms “protein”, “polypeptide”, and peptide refers to a polymer of at least 2 (e.g., at least 5) amino acids linked by a peptide bond. The term “polypeptide” does not denote a specific length of the polymer chain of amino acids. It is common in the art to refer to shorter polymers of amino acids (e.g., approximately 2-50 amino acids) as peptides; and to refer to longer polymers of amino acids (e.g., approximately over 50 amino acids) as polypeptides. However, the terms “peptide” and “polypeptide” and “protein” are used interchangeably herein. In some embodiments, the protein is folded into its three-dimensional structure. Where polypeptides (e.g., in a linear (i.e., primary) structure are contemplated herein, it should be understood that proteins folded into their three-dimensional structure (i.e., tertiary or quaternaryAttorney Docket No.63340.10WO01 structure) are also provided herein and vice versa. Proteins include e.g., naturally occurring proteins, variant (e.g., functional variants) of naturally occurring proteins, fragments (e.g., functional fragments) of naturally occurring proteins, and synthetic proteins (i.e., not naturally occurring proteins).

[0135] The terms “RNA” and “polyribonucleotide” are used interchangeably herein and refer to macromolecules that include multiple ribonucleotides that are polymerized via phosphodiester bonds. Ribonucleotides are nucleotides in which the sugar is ribose. RNA may contain modified nucleotides; and contain natural, non-natural, or altered internucleoside linkages, such as a phosphoroamidate linkage or a phosphorothioate linkage, instead of the phosphodiester found between the nucleotides of an unmodified nucleic acid molecule.

[0136] The term “scFv” or “single chain variable fragment” refers to an antibody that comprises a VH region operably connected via a peptide linker to a VL region, wherein the VH and VL regions associate to specifically bind an antigen (e.g., form an antigen binding domain). In some embodiments, the scFv comprises from N- to C-terminus an VH region, a peptide linker, and an VL region. In some embodiments, the scFv comprises from N- to C-terminus an VL region, a peptide linker, and an VH region.

[0137] The term “(scFv)2” as used herein refers to an antibody that comprises a first and a second scFv operably connected (e.g., via a peptide linker). The first and second scFv can specifically bind the same or different antigens. In some embodiments, the first and second scFv are operably connected by a peptide linker.

[0138] The term “scFv-Fc” as used herein refers to an antibody that comprises a scFv operably linked (e.g., via a peptide linker) to an Fc domain or subunit of an Fc domain. In some embodiments, a scFv is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first scFv is operably connected to a first Fc domain and a second scFv is operably connected to a second Fc domain of a first and second Fc domain pair.

[0139] The term “(scFv)2-Fc” as used herein refers to a (scFv)2 operably linked (e.g., via a peptide linker) to an Fc domain or a subunit of an Fc domain. In some embodiments, a (scFv)2is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first (scFv)2 is operably connected to a first Fc domain and a second (scFv)2 is operably connected to a second Fc domain of a first and second Fc domain pair.

[0140] As used herein, the term “single domain antibody” or “sdAb” refers to an antibodyAttorney Docket No.63340.10WO01 having a single monomeric variable antibody domain. A sdAb is able to specifically bind to a specific antigen. A VHH (as defined herein) is an example of a sdAb.

[0141] As used herein, the term “specifically binds” refers to preferential interaction, i.e., significantly higher binding affinity, between a first protein (e.g., an antibody) and a second protein (e.g., an antigen) relative to other amino acid sequences. Herein, when a first protein is said to “specifically bind” to a second protein, it is understood that the first protein specifically binds to an epitope of the second protein. The term “epitope” refers to the portion of the second protein that the first protein specifically recognizes. The term specifically binds includes molecules that are cross reactive with the same epitope of a different species. For example, an antibody that specifically binds human CLEC2D may be cross reactive with CLEC2D of another species (e.g., cynomolgus, murine, etc.), and still be considered herein to specifically bind human CLEC2D. A protein can specifically bind more than one different protein. Specific binding can be measured, e.g., through measuring binding affinity (e.g., using standard methods known in the art and described herein (e.g., surface plasmon resonance (SPR) (e.g., BIAcore®-based assay), a common method known in the art (see, e.g., Wilson, Science 295:2103, 2002; Wolff et al., Cancer Res. 55:2560, 1993; and U.S. Patent Nos. 5,283,173, 5,468,614, the full contents of each of which are incorporated by reference herein for all purposes).

[0142] As used herein, the term “subject” includes any animal, such as a human or other animal. In some embodiments, the subject is a vertebrate animal (e.g., mammal, bird, fish, reptile, or amphibian). In some embodiments, the subject is a human. In some embodiments, the method subject is a non-human mammal. In some embodiments, the subject is a non-human mammal is such as a non-human primate (e.g., monkeys, apes), ungulate (e.g., cattle, buffalo, sheep, goat, pig, camel, llama, alpaca, deer, horses, donkeys), carnivore (e.g., dog, cat), rodent (e.g., rat, mouse), or lagomorph (e.g., rabbit). In some embodiments, the subject is a bird, such as a member of the avian taxa Galliformes (e.g., chickens, turkeys, pheasants, quail), Anseriformes (e.g., ducks, geese), Paleaognathae (e.g., ostriches, emus), Columbiformes (e.g., pigeons, doves), or Psittaciformes (e.g., parrots).

[0143] As used herein, the term “therapeutic agent” refers to an agent (e.g., a conjugate described herein) capable of achieving a desired therapeutic result in a subject or ex vivo (e.g., capable of treating a disease as defined herein) when administered at a therapeutically effective amount.Attorney Docket No.63340.10WO01

[0144] As used herein, the term “therapeutically effective amount” of a therapeutic agent refers to any amount of the therapeutic agent that, when used alone or in combination with another therapeutic agent, improves a disease condition, e.g., protects a subject against the onset of a disease (or infection) (prophylaxis); improves a symptom of disease or infection, e.g., decreases severity of disease or infection symptoms, decreases frequency or duration of disease or infection symptoms, increases disease or infection symptom-free periods; prevents or reduces impairment or disability due to the disease or infection; or promotes disease (or infection) regression. The ability of a therapeutic agent to improve a disease condition can be evaluated using a variety of methods known to the skilled practitioner, such as in human subjects during clinical trials, in animal model systems predictive of efficacy in humans, or by assaying the activity of the agent in in vitro assays.

[0145] As used herein, the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disease and / or symptom(s) associated therewith or obtaining a desired pharmacologic and / or physiologic effect. It will be appreciated that, although not precluded, treating a disease does not require that the disease, or symptom(s) associated therewith be completely eliminated. In some embodiments, the effect is therapeutic, i.e., without limitation, the effect partially or completely reduces, diminishes, abrogates, abates, alleviates, decreases the intensity of, or cures a disease and / or adverse symptom attributable to the disease. In some embodiments, the effect is preventative (i.e., prophylactic), i.e., the effect protects or prevents an occurrence or reoccurrence of a disease. To this end, the presently disclosed methods comprise administering a therapeutically effective amount of e.g., a conjugate described herein (or a carrier, pharmaceutical composition, etc. comprising the same).

[0146] As used herein, the term “toll-like receptor 9” or “TLR9” refers to the toll-like receptor protein that, e.g., expressed on endosomes of e.g., B cells myeloid cells, internalized from the plasma membrane and binds e.g., CpG ODNs. The amino acid sequence of a reference mature (no signal sequence) human TLR9 protein is set forth in SEQ ID NO: 154 (UniProt Ref.: Q9NR96).

[0147] As used herein, the term “variation” or “variant” or use the like in reference to a nucleotide or nucleic acid sequence refers to a nucleic acid molecule that comprises at least one substitution, addition, deletion, or inversion of one or more nucleotide compared to a reference nucleic acid molecule. Likewise, as used herein, the term “variation” or “variant” or use the like with reference to a peptide or protein refers to a peptide or protein that comprises at least oneAttorney Docket No.63340.10WO01 substitution, addition, deletion, or inversion of an amino acid residue compared to a reference peptide or protein.

[0148] A “variation that promotes heterodimerization of a first Fc region and a second Fc region” (or similar phrasing) is a manipulation of the peptide backbone or the post-translational modifications of an Fc region that reduces or prevents the association of a polypeptide comprising the Fc region with an identical polypeptide to form a homodimer. A modification promoting association as used herein particularly includes separate modifications made to each of the two Fc regions desired to associate (i.e., a first Fc region and a second Fc region), wherein the modifications are complementary to each other so as to promote association of the two Fc regions. For example, a modification promoting association may alter the structure or charge of one or both of the Fc regions so as to make their association sterically or electrostatically favorable, respectively. Thus, heterodimerization occurs between a polypeptide comprising the first Fc region and a polypeptide comprising the second Fc region, which might be non-identical in the sense that further components fused to each of the Fc regions (e.g., antigen binding domains) are not the same. In some embodiments the modification promoting association comprises an amino acid mutation in the Fc region, specifically an amino acid substitution. In a particular embodiment, the modification promoting association comprises a separate amino acid mutation, specifically one or more amino acid substitution, in each of the first Fc region and the second Fc region. See, e.g., § 5.3.3.2.

[0149] As used herein, the term “variable region” refers to a portion of an antibody, generally, a portion of a light or heavy chain, typically about the amino-terminal 110 to 120 amino acids or 110 to 125 amino acids in the mature heavy chain and about 90 to 115 amino acids in the mature light chain, which differ extensively in sequence among antibodies and are used in the binding and specificity of a particular antibody for its particular antigen. The variability in sequence is concentrated in those regions called complementarity determining regions (CDRs) while the more highly conserved regions in the variable domain are called framework regions (FR). Without wishing to be bound by any particular mechanism or theory, it is believed that the CDRs of the light and heavy chains are primarily responsible for the interaction and specificity of the antibody with antigen. In certain embodiments, the variable region is a human variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and human framework regions (FRs). In particular embodiments, the variable region is a primate (e.g., non-humanAttorney Docket No.63340.10WO01 primate) variable region. In certain embodiments, the variable region comprises rodent or murine CDRs and primate (e.g., non-human primate) framework regions (FRs).

[0150] The terms “VL” and “VL region” are used interchangeably to refer to an immunoglobulin light chain variable region. A VL region can be incorporated into an antibody, e.g., a scFv, a Fab, a full-length antibody. For example, a scFv comprises a VL region operably connected via a peptide linker to a VH region.

[0151] The terms “VH” and “VH region” are used interchangeably to refer to an immunoglobulin heavy chain variable region. A VH region can be incorporated into an antibody, e.g., a scFv, a Fab, a full-length antibody. For example, a scFv comprises a VH region operably connected via a peptide linker to a VL region.

[0152] The term “VHH” as used herein refers to a type of single domain antibody (sdAb) that has a single monomeric heavy chain variable antibody domain (VH). Such antibodies can be found in or produced from camelid mammals (e.g., camels, llamas) which are naturally devoid of light chains or synthetically produced.

[0153] The term “(VHH)2” as used herein refers to an antibody that comprises a first and a second VHH operably connected (e.g., via a peptide linker). The first and the second VHH can specifically bind the same or different antigens. In some embodiments, the first and second VHH are operably connected by a peptide linker.

[0154] The term “VHH-Fc” as used herein refers to an antibody that comprises a VHH operably linked (e.g., via a peptide linker) to an Fc domain or a subunit of an Fc domain. In some embodiments, a VHH is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first VHH is operably connected to a first Fc domain and a second VHH is operably connected to a second Fc domain of a first Fc and a second Fc pair.

[0155] The term “(VHH)2-Fc” as used herein refers to (VHH)2operably linked (e.g., via a peptide linker) to an Fc domain or a subunit of an Fc domain. In some embodiments, a (VHH)2 is operably connected to only a first Fc domain of a first and a second Fc domain pair. In some embodiments, a first (VHH)2is operably connected to a first Fc domain and a second (VHH)2is operably connected to a second Fc domain of a first Fc and a second Fc pair. 5.2 Conjugates

[0156] Provided herein are, inter alia, conjugates (e.g., anti-CLEC2D antibody-CpG ODNAttorney Docket No.63340.10WO01 conjugates), useful in, inter alia, agonizing TLR9 in CLEC2D expressing immune cells (e.g., myeloid cells, B cells) and treating cancer (e.g., solid tumors). The conjugates described herein comprise a targeting agent (e.g., a CLEC2D targeting agent (e.g., an anti-CLEC2D antibody described herein)) and (e.g., operably connected to) a CpG ODN molecular payload (e.g., a CpG ODN described herein). 5.3 CLEC2D Targeting Agents

[0157] As described above, the conjugates described herein comprise a CLEC2D targeting agent (e.g., for targeting a molecular payload (e.g., a CpG ODN described herein)) to CLEC2D expressing cells (e.g., B cells, myeloid cells) (e.g., within a subject (e.g., within the tumor microenvironment with a subject))).

[0158] CLEC2D is a C type lectin receptor that, inter alia, regulates immune responses through, e.g., binding to the type II transmembrane C-type lectin-like receptor, CD161 (known to be expressed by, e.g., natural killer (NK) cells). Multiple isoforms of hCLEC2D are known produced by alternative splicing, with isoform 1 being the only isoform predominantly expressed at the cell surface. The amino acid sequence of a reference hCLEC2D isoform 1 protein is set forth in SEQ ID NO: 1. The amino acid sequence of a reference hCD161 protein is set forth in SEQ ID NO: 2. See Table 1, herein. Table 1. The Amino Acid Sequence of a Reference hCLEC2D and hCD161 Protein. Description Amino Acid SequenceSEQ IDNOAttorney Docket No.63340.10WO01 MERLWGLFQRAQQLSPRSSQTVYQRVEGPRKGHLEEEEEDGEEGAETL AHFCPMELRGPEPLGSRPRQPNLIPWAAAGRRAAPYLVLTALLIFTGA FLLGYVAFRGSCQACGDSVLVVSEDVNYEPDLDFHQGRLYWSDLQAMFing agents) may be used in accordance with the present disclosure. For example, the targeting agent (e.g., the CLEC2D targeting agent) may comprise (or consist of) a small molecule, an oligonucleotide (e.g., DNA, RNA, RNA / DNA hybrid), a protein (e.g., an antibody, a peptide), a lipid (e.g., a microvesicle), or a carbohydrate (e.g., a polysaccharide). In some embodiments, the targeting agent is a protein. In some embodiments, the targeting agent is an antibody. In some embodiments, the targeting agent is an antibody-like scaffold (e.g., as described herein). Exemplary targeting agents are described in further detail herein, however, it should be appreciated that the exemplary targeting agents provided herein are not meant to be limiting.

[0160] In some embodiments, the CLEC2D targeting agent specifically binds to hCLEC2D. In some embodiments, the CLEC2D targeting agent (e.g., an anti-CLEC2D antibody) specifically binds hCLEC2D and one or more of murine CLEC2D, rat CLEC2D, a non-human primate CLEC2D (e.g., cynomolgus CLEC2D).

[0161] In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent enhances the distribution and / or uptake (e.g., into a cell, e.g., into a cell in a subject, e.g., a cell that expresses CLEC2D (e.g., hCLEC2D)) (e.g., an immune cell (e.g., a B cell, a myeloid cell (e.g., a macrophage, dendritic cell (e.g., a plasmacytoid dendritic cell, a classical dendritic cell)))) of a molecular payload (e.g., a CpG ODN described herein) (e.g., as compared to molecular payload (e.g., a CpG ODN described herein) that lacks the targeting moiety). In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent alters (e.g., extends) the lifetime (e.g., in vivo) of theAttorney Docket No.63340.10WO01 molecular payload (e.g., a CpG ODN described herein) (e.g., as compared to molecular payload (e.g., a CpG ODN described herein) that lacks the targeting moiety). In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent provides an enhanced affinity for a selected target, e.g., a selected cell type, compartment (e.g., cell type, tissue, organ or region of the body) (e.g., as compared to molecular payload (e.g., a CpG ODN described herein) that lacks the targeting moiety) (e.g., an immune cell (e.g., a B cell, a myeloid cell (e.g., a macrophage, dendritic cell (e.g., a plasmacytoid dendritic cell, a classical dendritic cell)))). In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent inhibits (partially, substantially, or fully) binding of CLEC2D to CD161 (e.g., hCD161).

[0162] In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent is a protein. In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent comprises an antibody-like scaffold. In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent is an anti-CLEC2D antibody (e.g., an anti-CLEC2D antibody). In some embodiments, the antibody comprises or consists of a full-length antibody, Fab, Fab', F(ab')2, Fab-Fc, scFv, scFv-Fc, (scFv)2-Fc, Fv, a single domain antibody (sdAb) (e.g., a VHH), a sdAb-Fc (e.g., a VHH-Fc), (sdAb)2 (e.g., a (VHH)2, or a (sdAb)2-Fc (e.g., (VHH)2-Fc). In some embodiments, the antibody comprises or consists of a full-length antibody, Fab, Fab', F(ab')2, Fab-Fc, scFv, scFv-Fc, (scFv)2-Fc, sdAb-Fc (e.g., a VHH-Fc), or (sdAb)2-Fc (e.g., (VHH)2-Fc). In some embodiments, the antibody comprises or consists of a full-length antibody. In some embodiments, the antibody comprises or consists of a Fab. In some embodiments, the antibody comprises or consists of a F(ab')2. In some embodiments, the antibody comprises or consists of a Fab-Fc. In some embodiments, the antibody comprises or consists of a scFv-Fc. In some embodiments, the antibody comprises or consists of a (scFv)2-Fc. In some embodiments, the antibody comprises or consists of a sdAb-Fc (e.g., a VHH- Fc). In some embodiments, the antibody comprises or consists of a o(sdAb)2-Fc (e.g., (VHH)2-Fc)

[0163] In some embodiments, the antibody is an IgG1, IgG2, IgG3, or IgG4 antibody. In some embodiments, the antibody is an IgG1 or IgG4 antibody. In some embodiments, the antibody is an IgG1 antibody. In some embodiments, the antibody is an IgG4 antibody. In some embodiments, the antibody is a hIgG1, hIgG2, hIgG3, or hIgG4 antibody. In some embodiments, the antibody is a hIgG1 or hIgG4 antibody. In some embodiments, the antibody is a hIgG1 antibody. In some embodiments, the antibody is a hIgG4 antibody. 5.3.1 Exemplary Anti-CLEC2D AntibodiesAttorney Docket No.63340.10WO01

[0164] In some embodiments, the CLEC2D (e.g., hCLEC2D) targeting agent comprises an anti-CLEC2D (e.g., hCLEC2D) antibody. Anti-CLEC2D (e.g., hCLEC2D) antibodies that can be employed in the conjugates described herein are known in the art. For example, exemplary anti- CLEC2D (e.g., hCLEC2D) antibodies known in the art that can be employed in the conjugates described herein include, but are not limited to, e.g., C5511 and C4608 (see, e.g., US20200291120), and those described in US9127052 and US20200291120 (see, e.g., Tables 2, 4, 6, and 9A); the entire contents of each of which is incorporated herein by reference for all purposes.

[0165] The amino acid sequence of exemplary anti-hCLEC2D antibodies that can be utilized in the conjugates described herein is provided in Table 2. The CDRs of the antibodies in Table 2, are denoted according to Kabat. A person of ordinary skill in the art would be able to determine the CDRs as defined by another scheme, e.g., Chothia, IMGT, using ordinary methods known in the art. Table 2. The Amino Acid Sequence of Exemplary Anti-hCLEC2D Antibodies. DescriptionSEQID NOAmino Acid Sequence K L A Y K L T L P D R T A N A Y TAttorney Docket No.63340.10WO01 ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC HC – QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGK L T L P D R T A N K L A Y K L A Y K L A YAttorney Docket No.63340.10WO01 VH CDR2 204VIEYDGSNKYYADSVKGVH CDR3 205DYGLYSLAAWDYALLDYVL CDR1 6K L A Y K L A Y K L A Y K L A YAttorney Docket No.63340.10WO01 VL CDR1 200HASQSISSYLNVL CDR2 201AASLLQSVL CDR3 202K L A Y K L A Y K L A Y K L A YAttorney Docket No.63340.10WO01 VL CDR3 226QQANSFLTVH EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGK 227 GLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSL A Y K L A Y P S A Y K L A YAttorney Docket No.63340.10WO01 VH QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYAMHWVRQAPGK 245 GLEWVAVISYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSL RAEDTAVYYCARDGDDYRYDYYSYGFDVWGQGTLVTVSS A Y Q L A Y K L A Y Q L A YAttorney Docket No.63340.10WO01 VH EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGK 263 GLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSL RAEDTAVYYCARDLEAWTTVYGSYGFDVWGQGTLVTVSS A Y K N A Y K N A Y K L A YAttorney Docket No.63340.10WO01 VH QVQLVESGGGVVQPGRSLRLSCAASGFTFSDYAMHWVRQAPGK 148 GLEWVAVIEYDGSNKYYADSVKGRFTISRDNSKNTLYLQMNSL RAEDTAVYYCARDYGLYSLAAWDYALLDYWGQGTLVTVSS A Y K Q Q L Q E K T A YAttorney Docket No.63340.10WO01 VH QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGK 167 GLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVT AADTAVYYCARGRPAPSWVKTRNWFDPWGQGTLVTVSS A Y K L Q E K T V A Y K N Q V

[0166] In some embodiments, the CLEC2D binding protein comprises a VH that comprises: a VH CDR1, a VH CDR2, and a VH CDR3. In some embodiments, the CLEC2D binding protein comprises a VL that comprises: a VL CDR1, a VL CDR2, and a VL CDR3.

[0167] In some embodiments, the CLEC2D binding protein comprises a VH that comprises: aAttorney Docket No.63340.10WO01 VH CDR1, a VH CDR2, and a VH CDR3; and a VL that comprises: a VL CDR1, a VL CDR2, and a VL CDR3.

[0168] In some embodiments, the CLEC2D binding protein comprises a CLEC2D binding protein provided in Table 2.

[0169] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0170] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition,Attorney Docket No.63340.10WO01 etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0171] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH set forth in Table 2; the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH set forth in Table 2; the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH set forth in Table 2; the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL set forth in Table 2; the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL set forth in Table 2; and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL set forth in Table 2.

[0172] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0173] In some embodiments, the amino acid sequence of VH CDR1 consists of the aminoAttorney Docket No.63340.10WO01 acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0174] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH set forth in Table 2, or the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH set forth in Table 2, or the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR2 of a VL set forth in Table 2Attorney Docket No.63340.10WO01 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL set forth in Table 2, or the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0175] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH set forth in Table 2; the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH set forth in Table 2; the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH set forth in Table 2; the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL set forth in Table 2; the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL set forth in Table 2; and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL set forth in Table 2.

[0176] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid of a VH CDR1 of a VH set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of a VH set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of a VL set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0177] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VHAttorney Docket No.63340.10WO01 comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises the amino acid sequence of a VL set forth in Table 2.

[0178] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL consists of the amino acid sequence of a VL set forth in Table 2.

[0179] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the aminoAttorney Docket No.63340.10WO01 acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acid sequence of a VL CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0180] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion,Attorney Docket No.63340.10WO01 addition, etc.).

[0181] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 set forth in Table 2; the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 set forth in Table 2; the amino acid sequence of VH CDR3 comprises of a VH CDR3 set forth in Table 2; the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 set forth in Table 2; the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 set forth in Table 2; and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 set forth in Table 2.

[0182] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0183] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acidAttorney Docket No.63340.10WO01 sequence of a VL CDR2 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0184] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 set forth in Table 2, or the amino acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 set forth in Table 2, or the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 set forth in Table 2, or the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 set forth in Table 2, or the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 set forth in Table 2, or the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 set forth in Table 2, or the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0185] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 set forth in Table 2; the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 set forth in Table 2; the amino acid sequence of VH CDR3 consists of a VH CDR3 set forth in Table 2; the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 set forth in Table 2; the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 set forth in Table 2; and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 set forth in Table 2.

[0186] In some embodiments, the amino acid sequence of VH CDR1 consists of the aminoAttorney Docket No.63340.10WO01 acid of a VH CDR1 set forth in Table 2, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 set forth in Table 2 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0187] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL set forth in Table 2. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence of a VH set forth in Table 2; and the amino acid sequence of the VL comprises the amino acid sequence of a VL set forth in Table 2.

[0188] In some embodiments, the anti-CLEC2D (e.g., hCLEC2D) antibody comprises an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0189] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / orAttorney Docket No.63340.10WO01 incorporated by reference herein, or the amino acid of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of a VL of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0190] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH of an anti-Attorney Docket No.63340.10WO01 CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0191] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR3 comprises of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2Attorney Docket No.63340.10WO01 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0192] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0193] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of a VH of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, orAttorney Docket No.63340.10WO01 the amino acid sequence of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of a VL of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0194] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of a VH of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of a VH of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL of an anti-Attorney Docket No.63340.10WO01 CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of a VL of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0195] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR3 consists of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0196] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid of a VH CDR1 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of a VH of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprisingAttorney Docket No.63340.10WO01 no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0197] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, theAttorney Docket No.63340.10WO01 amino acid sequence of the VH comprises the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0198] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL consists of the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0199] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising 1, 2, or 3 aminoAttorney Docket No.63340.10WO01 acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0200] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution,Attorney Docket No.63340.10WO01 deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0201] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR2 comprises the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR3 comprises of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR1 comprises the amino acid sequence of a VL CDR1 of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR2 comprises the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of VL CDR3 comprises the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0202] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acidAttorney Docket No.63340.10WO01 of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0203] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / orAttorney Docket No.63340.10WO01 incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0204] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of a VH CDR3 of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / orAttorney Docket No.63340.10WO01 incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, or the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0205] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR2 consists of the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VH CDR3 consists of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR1 consists of the amino acid sequence of a VL CDR1 of an anti- CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; the amino acid sequence of VL CDR2 consists of the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of VL CDR3 consists of the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0206] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid of a VH CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein, comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VH CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR1 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of a VL CDR2 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution,Attorney Docket No.63340.10WO01 deletion, addition, etc.); and the amino acid sequence of a VL CDR3 of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0207] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence of a VH of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein; and the amino acid sequence of the VL comprises the amino acid sequence of a VL of an anti-CLEC2D (e.g., hCLEC2D) antibody named and / or incorporated by reference herein.

[0208] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations (e.g.,Attorney Docket No.63340.10WO01 substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0209] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0210] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4; the amino acid sequence of VH CDR3 comprises theAttorney Docket No.63340.10WO01 amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8.

[0211] In some embodiments, the amino acid sequence of VH CDR1 the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0212] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO:Attorney Docket No.63340.10WO01 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0213] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6, or the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7, or the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8, or the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0214] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4; the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6; the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8.

[0215] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3Attorney Docket No.63340.10WO01 consists of the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 6 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 7 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 8 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0216] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises the amino acid sequence set forth in SEQ ID NO: 10.

[0217] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH consistsAttorney Docket No.63340.10WO01 of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 10.

[0218] In some embodiments, the amino acid sequence of the heavy chain comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the heavy chain comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the heavy chain comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the heavy chain comprises the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain comprises the amino acid sequence set forth in SEQ ID NO: 12.

[0219] In some embodiments, the amino acid sequence of the heavy chain consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and theAttorney Docket No.63340.10WO01 amino acid sequence of the light chain consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the heavy chain consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the heavy chain consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 12. In some embodiments, the amino acid sequence of the heavy chain consists of the amino acid sequence set forth in SEQ ID NO: 11; and the amino acid sequence of the light chain consists of the amino acid sequence set forth in SEQ ID NO: 12.

[0220] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 193, or the amino acid sequence set forth in SEQ ID NO: 193 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 194, or the amino acid sequence set forth in SEQ ID NO: 194 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0221] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acidAttorney Docket No.63340.10WO01 sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 193, or the amino acid sequence set forth in SEQ ID NO: 193 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 194, or the amino acid sequence set forth in SEQ ID NO: 194 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0222] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4; the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 193; the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 194; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195.

[0223] In some embodiments, the amino acid sequence of VH CDR1 the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 193 comprising no more than 1, 2, orAttorney Docket No.63340.10WO01 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 194 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0224] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 193, or the amino acid sequence set forth in SEQ ID NO: 193 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 194, or the amino acid sequence set forth in SEQ ID NO: 194 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0225] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4, or the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5, or the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 193, or the amino acid sequence set forth in SEQ ID NO:Attorney Docket No.63340.10WO01 193 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 194, or the amino acid sequence set forth in SEQ ID NO: 194 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0226] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4; the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5; the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 193; the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 194; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195.

[0227] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 4 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 5 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 193 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 194 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0228] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acidAttorney Docket No.63340.10WO01 sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 196. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 196. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 196. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL comprises the amino acid sequence set forth in SEQ ID NO: 196.

[0229] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 196. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 196. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 196. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NO: 9; and the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 196.Attorney Docket No.63340.10WO01

[0230] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 204, or the amino acid sequence set forth in SEQ ID NO: 204 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 205, or the amino acid sequence set forth in SEQ ID NO: 205 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 197, or the amino acid sequence set forth in SEQ ID NO: 197 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 198, or the amino acid sequence set forth in SEQ ID NO: 198 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0231] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 204, or the amino acid sequence set forth in SEQ ID NO: 204 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 205, or the amino acid sequence set forth in SEQ ID NO: 205 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 197, or the amino acid sequence set forth in SEQ ID NO: 197 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 198, or the amino acid sequence set forth in SEQ ID NO: 198 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195, or theAttorney Docket No.63340.10WO01 amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0232] In some embodiments, the amino acid sequence of VH CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 204; the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 205; the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 197; the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 198; and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195.

[0233] In some embodiments, the amino acid sequence of VH CDR1 the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 204 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 205 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 comprises the amino acid sequence set forth in SEQ ID NO: 197 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 comprises the amino acid sequence set forth in SEQ ID NO: 198 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 comprises the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0234] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 204, or the amino acid sequence set forth in SEQ ID NO: 204 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 205, or the amino acid sequence set forth in SEQ ID NO: 205 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ IDAttorney Docket No.63340.10WO01 NO: 197, or the amino acid sequence set forth in SEQ ID NO: 197 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 198, or the amino acid sequence set forth in SEQ ID NO: 198 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0235] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3, or the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 204, or the amino acid sequence set forth in SEQ ID NO: 204 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 205, or the amino acid sequence set forth in SEQ ID NO: 205 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 197, or the amino acid sequence set forth in SEQ ID NO: 197 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 198, or the amino acid sequence set forth in SEQ ID NO: 198 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195, or the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0236] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3; the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 204; the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 205; the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 197; the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 198; and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195.Attorney Docket No.63340.10WO01

[0237] In some embodiments, the amino acid sequence of VH CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 3 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 204 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VH CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 205 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR1 consists of the amino acid sequence set forth in SEQ ID NO: 197 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); the amino acid sequence of VL CDR2 consists of the amino acid sequence set forth in SEQ ID NO: 198 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.); and the amino acid sequence of VL CDR3 consists of the amino acid sequence set forth in SEQ ID NO: 195 comprising no more than 1, 2, or 3 amino acid variations (e.g., substitution, deletion, addition, etc.).

[0238] In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 199. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL comprises an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 199. In some embodiments, the amino acid sequence of the VH comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL comprises an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 199. In some embodiments, the amino acid sequence of the VH comprises the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL comprises the amino acid sequence set forth in SEQ ID NO: 199.Attorney Docket No.63340.10WO01

[0239] In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 199. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL consists of an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 199. In some embodiments, the amino acid sequence of the VH consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL consists of an amino acid sequence at least 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence set forth in SEQ ID NO: 199. In some embodiments, the amino acid sequence of the VH consists of the amino acid sequence set forth in SEQ ID NO: 210; and the amino acid sequence of the VL consists of the amino acid sequence set forth in SEQ ID NO: 199. 5.3.2 Exemplary Properties of CLEC2D Binding Proteins 5.3.2.1 Affinity of CLEC2D Targeting Moiety

[0240] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a dissociation constant (Kd) for CLEC2D (e.g., hCLEC2) binding of < 10 μΜ, < 100 μΜ , < 10 μΜ , < 1 μΜ, < 100 nΜ, < 10 nΜ, < 1 nΜ, < 0.1 nΜ, < 0.01 nΜ, or < 0.001 nΜ, and / or > 0.01 μΜ, 0.1 μΜ, or 1 μΜ (e.g. , 10-5M or less, 10-6 M or less, 10-8 M or less, e.g., from 1 μΜ to 10 μΜ, e.g., from 0.1 μΜ to 10 μΜ, e.g., from 10-6 M to 10-9 M, e.g., from 10-8 M to 10-13 M, e.g., from 10-9 M to 10-13 M) (e.g., as measured by SPR).

[0241] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of < 1 nΜ (e.g., as measured by SPR). In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of less than 1 nM (e.g., as measured by SPR). In some embodiments, the CLEC2D targeting moiety (e.g., anti-Attorney Docket No.63340.10WO01 CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of from about 0.5nM - 1 nM (e.g., as measured by SPR).

[0242] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of < 1 nΜ (e.g., as measured according to the methods in Example 2). In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of less than 1 nM (e.g., as measured according to the methods in Example 2). In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of from about 0.5nM - 1 nM (e.g., as measured according to the methods in Example 2).

[0243] Binding affinity can be measured by standard assays known in the art. For example, binding affinity can be measured by surface plasmon resonance (SPR) (e.g., BIAcore®-based assay), a common method known in the art (see, e.g., Wilson, Science 295:2103, 2002; Wolff et al., Cancer Res. 55:2560, 1993; and U.S. Patent Nos. 5,283,173, 5,468,614, the full contents of each of which are incorporated by reference herein for all purposes). SPR measures changes in the concentration of molecules at a sensor surface as molecules bind to or dissociate from the surface. The change in the SPR signal is directly proportional to the change in mass concentration close to the surface, thereby allowing measurement of binding kinetics between two molecules (e.g., proteins). The dissociation constant for the complex can be determined by monitoring changes in the refractive index with respect to time as buffer is passed over the chip. In some embodiments, the affinity of a CLEC2D binding protein for CLEC2D (e.g., KD) is measured by SPR.

[0244] Other suitable assays for measuring the binding affinity include, for example, immunoassays such as enzyme linked immunosorbent assays (ELISA) and radioimmunoassays (RIA), or determination of binding by monitoring the change in the spectroscopic or optical properties of the proteins through fluorescence, UV absorption, circular dichroism, or nuclear magnetic resonance (NMR). Other exemplary assays include, but are not limited to, Western blot, analytical ultracentrifugation, spectroscopy, flow cytometry, sequencing and other methods for detection of binding of proteins. 5.3.2.2 Inhibition of CD161 Binding to CLEC2D

[0245] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D toAttorney Docket No.63340.10WO01 CD161. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) partially inhibits binding of CLEC2D to CD161. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) substantially inhibits binding of CLEC2D to CD161. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) fully inhibits binding of CLEC2D to CD161.

[0246] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of hCLEC2D to hCD161. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) partially inhibits binding of hCLEC2D to hCD161. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) substantially inhibits binding of hCLEC2D to hCD161. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) fully inhibits binding of hCLEC2D to hCD161.

[0247] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D and exhibits an IC50 of < 10 μΜ, < 100 μΜ , < 10 μΜ , < 1 μΜ, < 100 nΜ, < 10 nΜ, < 1 nΜ, < 0.1 nΜ, < 0.01 nΜ, or < 0.001 nΜ, and / or > 0.01 μΜ, 0.1 μΜ, or 1 μΜ (e.g. , 10-5M or less, 10-6 M or less, 10-8 M or less, e.g., from 1 μΜ to 10 μΜ, e.g., from 0.1 μΜ to 10 μΜ, e.g., from 10-6 M to 10-9 M, e.g., from 10-8 M to 10-13 M, e.g., from 10-9 M to 10-13 M) (e.g., as measured by SPR).

[0248] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D and exhibits an IC50 of < 10 nΜ. In some embodiments, the CLEC2D targeting moiety (e.g., anti- CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D and exhibits an IC50 of less than 5 nM. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D and exhibits an IC50 of less than 2 nM. In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of from about 0.5nM - 2 nM.

[0249] In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D andAttorney Docket No.63340.10WO01 exhibits an IC50 of < 10 nΜ (e.g., as measured according to the methods in Example 2). In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D and exhibits an IC50 of less than 5 nM (e.g., as measured by SPR). In some embodiments, the CLEC2D targeting moiety (e.g., anti-CLEC2D antibody) (e.g., described herein)) inhibits (e.g., partially, substantially, or fully) binding of CLEC2D and exhibits an IC50 of less than 2 nM (e.g., as measured according to the methods in Example 2). In some embodiments, the CLEC2D targeting moiety (e.g., anti- CLEC2D antibody) (e.g., described herein)) exhibits a Kd for CLEC2D (e.g., hCLEC2) binding of from about 0.5nM - 2 nM (e.g., as measured according to the methods in Example 2).

[0250] Standard binding assays to measure binding of CD161 and CLEC2D are known in the art and described herein. See, e.g., Wade M, Méndez J, Coussens NP, et al. Inhibition of Protein- Protein Interactions: Cell-Based Assays. 2017 Nov 20. In: Markossian S, Grossman A, Brimacombe K, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. Available from: https: / / www.ncbi.nlm.nih.gov / books / NBK464632 / ; Arkin MR, Glicksman MA, Fu H, et al. Inhibition of Protein-Protein Interactions: Non-Cellular Assay Formats. 2012 Mar 18 [Updated 2012 Oct 1]. In: Markossian S, Grossman A, Brimacombe K, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences; 2004-. Available from: https: / / www.ncbi.nlm.nih.gov / books / NBK92000 / ; the entire contents of each of which are incorporated herein by reference for all purposes. For example, non-cell-based assays include, but are not limited to, ELISA. For further example, cell- based assays include, but are not limited to, energy transfer (Förster resonance energy transfer and bioluminescence resonance energy transfer) and protein complementation (fluorescence or enzymatic, e.g., luciferase). 5.3.3 Ig Constant Regions

[0251] In some embodiments, the antibody comprises an IgG CH2 region and an IgG CH3 region. In some embodiments, the antibody comprises a partial IgG hinge region, IgG CH2 region, and IgG CH3 region. In some embodiments, the antibody comprises of an IgG hinge region, IgG CH2 region, and IgG CH3 region. In some embodiments, the antibody comprises an IgG1 CH2 region and an IgG1 CH3 region. In some embodiments, the antibody comprises a partial IgG1Attorney Docket No.63340.10WO01 hinge region, IgG1 CH2 region, and IgG1 CH3 region. In some embodiments, the antibody comprises an IgG1 hinge region, IgG1 CH2 region, and IgG1 CH3 region. In some embodiments, the antibody comprises an IgG4 CH2 region and an IgG4 CH3 region. In some embodiments, the antibody comprises a partial IgG4 hinge region, IgG4 CH2 region, and IgG4 CH3 region. In some embodiments, the antibody comprises an IgG4 hinge region, IgG4 CH2 region, and IgG4 CH3 region.

[0252] In some embodiments, the antibody comprises an Ig Fc region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Ig Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the Ig Fc region comprises or consists of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of an IgG1 hinge region, an IgG1 CH2 region, and an IgG1 CH3 region. In some embodiments, the Ig Fc region comprises or consists of an IgG1 hinge region, an IgG1 CH2 region, and an IgG1 CH3 region. In some embodiments, the Ig Fc region comprises or consists of at least a portion of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region. In some embodiments, the Ig Fc region comprises or consists of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region.

[0253] In some embodiments, the antibody comprises a first Ig Fc region and a second Ig Fc region. In some embodiments, the first and / or second Ig Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of at least a portion of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of an IgG hinge region, an IgG CH2 region, and an IgG CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of at least a portion of an IgG1 hinge region, an IgG1 CH2 region, and an IgG1 CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of an IgG1 hinge region, an IgG1 CH2 region, and an IgG1 CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of at least a portion of an IgG4 hinge region, anAttorney Docket No.63340.10WO01 IgG4 CH2 region, and an IgG4 CH3 region. In some embodiments, the first and / or second Ig Fc region comprises or consists of an IgG4 hinge region, an IgG4 CH2 region, and an IgG4 CH3 region.

[0254] In some embodiments, the antibody comprises one or more hIg heavy chain constant regions (e.g., a CH2 region, a CH3 region, a hinge region, an Fc region). In some embodiments, the hIg is a human IgG (hIgG). In some embodiments, the hIgG is hIgG1, IgG2, IgG3, or IgG4. In some embodiments, the hIgG is IgG1 or IgG4. In some embodiments, the hIgG is hIgG1. In some embodiments, the hIgG is hIgG4.

[0255] In some embodiments, the antibody comprises a hIgG CH2 region and a hIgG CH3 region. In some embodiments, the antibody comprises a partial hIgG hinge region, hIgG CH2 region, and hIgG CH3 region. In some embodiments, the antibody comprises a hIgG hinge region, hIgG CH2 region, and hIgG CH3 region. In some embodiments, the antibody comprises a hIgG1 CH2 region and a hIgG1 CH3 region. In some embodiments, the antibody comprises a partial hIgG1 hinge region, hIgG1 CH2 region, and hIgG1 CH3 region. In some embodiments, the antibody comprises a hIgG1 hinge region, hIgG1 CH2 region, and hIgG1 CH3 region. In some embodiments, the antibody comprises a hIgG4 CH2 region and a hIgG4 CH3 region. In some embodiments, the antibody comprises a partial hIgG4 hinge region, hIgG4 CH2 region, and hIgG4 CH3 region. In some embodiments, the antibody comprises a hIgG4 hinge region, hIgG4 CH2 region, and hIgG4 CH3 region.

[0256] In some embodiments, the antibody comprises a hIg Fc region. In some embodiments, the hIg Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the hIg Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the hIg Fc region comprises or consists of at least a portion of a hIgG hinge region, a hIgG CH2 region, and a hIgG CH3 region. In some embodiments, the hIg Fc region comprises or consists of a hIgG hinge region, a hIgG CH2 region, and a hIgG CH3 region. In some embodiments, the hIg Fc region comprises or consists of at least a portion of a hIgG1 hinge region, a hIgG1 CH2 region, and a hIgG1 CH3 region. In some embodiments, the hIg Fc region comprises or consists of a hIgG1 hinge region, a hIgG1 CH2 region, and a hIgG1 CH3 region. In some embodiments, the hIg Fc region comprises or consists of at least a portion of a hIgG4 hinge region, a hIgG4 CH2 region, and a hIgG4 CH3 region. In some embodiments, the hIg Fc region comprises or consists of a hIgG4 hinge region, a hIgG4 CH2Attorney Docket No.63340.10WO01 region, and a hIgG4 CH3 region.

[0257] In some embodiments, the antibody comprises a first hIg Fc region and a second hIg Fc region. In some embodiments, the first and / or second hIg Fc region comprises or consists of at least a portion of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of a hinge region, a CH2 region, and a CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of at least a portion of a hIgG hinge region, a hIgG CH2 region, and a hIgG CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of a hIgG hinge region, a hIgG CH2 region, and a hIgG CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of at least a portion of a hIgG1 hinge region, a hIgG1 CH2 region, and a hIgG1 CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of a hIgG1 hinge region, a hIgG1 CH2 region, and a hIgG1 CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of at least a portion of a hIgG4 hinge region, a hIgG4 CH2 region, and a hIgG4 CH3 region. In some embodiments, the first and / or second hIg Fc region comprises or consists of a hIgG4 hinge region, a hIgG4 CH2 region, and a hIgG4 CH3 region.

[0258] In some embodiments, the antibody comprises one or more Ig (e.g., hIg) light chain constant region (e.g., a hIg light chain kappa constant region (κCL) or a hIg light chain lambda constant region (λCL).

[0259] The amino acid sequence of exemplary reference hIgG1 and hIgG4 heavy chain and light chain constant regions, which can be incorporated in one or more of the embodiments described herein (e.g., anti-CLEC2D (e.g., hCLEC2D) antibodies), is provided in Table 3. Table 3. The Amino Acid Sequence of Exemplary hIg Heavy Chain and Light Chain Constant Regions and Components Thereof. Description Amino Acid SequenceSEQOAttorney Docket No.63340.10WO01 hIgG1 CH3 Region GQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWES NGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS VMHEALHNHYTQKSLSLSPGK33Attorney Docket No.63340.10WO01 DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGK hI G1 CH1 + Hin e ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSAttorney Docket No.63340.10WO01 hIgG4 Hinge Region + ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTC CH2 Region + CH3 VVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVS Re ion VLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQ, on, wherein the amino acid sequence of the one or more hIg constant region comprises or consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3. In some embodiments, the amino acid sequence of the one or more hIg constant region comprises or consists of an amino acid sequence set forth in Table 3.

[0261] In some embodiments, the amino acid sequence of the one or more hIg constant region comprises or consists of an amino acid sequence set forth in Table 3, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequenceAttorney Docket No.63340.10WO01 of the one or more hIg constant region comprises or consists of an amino acid sequence set forth in Table 3, comprising or consisting of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the one or more hIg constant region comprises or consists of an amino acid sequence set forth in Table 3, comprising or consisting of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of an amino acid sequence set forth in Table 3, comprising or consisting of about no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions).

[0262] In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of an amino acid sequence set forth in Table 3, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid substitutions. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of an amino acid sequence set forth in Table 3, comprising or consisting of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of an amino acid sequence set forth in Table 3, comprising or consisting of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of an amino acid sequence set forth in Table 3, comprising or consisting of about no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions.

[0263] In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 30-57. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57.

[0264] In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the aminoAttorney Docket No.63340.10WO01 acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, comprising or consisting at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, comprising or consisting about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, comprising or consisting of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., amino acid substitutions, deletions, or additions).

[0265] In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid substitutions. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, comprising or consisting at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, comprising or consisting about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. In some embodiments, the amino acid sequence of the one or more hIg constant regions comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 30-57, comprising or consisting of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. 5.3.3.1 Ig Effector Function

[0266] As described herein, in some embodiments, the antibody comprises an Fc region (see, e.g., § 5.3.3). In some embodiments, the Fc region of an antibody described herein exhibits a decrease in one or more Fc effector function relative to a reference (e.g., wild type) Fc region. Exemplary Fc effector functions include, but are not limited to, antibody dependent cellular cytotoxicity (ADCC), antibody dependent cellular phagocytosis (ADCP), complement dependent cytotoxicity (CDC), and binding affinity to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))).

[0267] Standard in vitro and / or in vivo assays known in the art can be conducted to evaluateAttorney Docket No.63340.10WO01 Fc effector function, including, any one or more of ADCC, CDC, ADCP, Fc receptor (e.g., Fcγ receptor) binding affinity, and C1q binding affinity.

[0268] For example, ADCC activity can be assessed utilizing standard (radioactive and non- radioactive) methods known in the art (see, e.g., WO2006 / 082515, WO2012 / 130831), the entire contents of each of which is incorporated by reference herein for all purposes). For example, ADCC activity can be assessed using a chromium-5 (51Cr) assay. Briefly,51Cr is pre-loaded into target cells, NK cells are added to the culture, and radioactivity in the cell culture supernatant is assessed (indicative of lysis of the target cells by the NK cells). Similar non-radioactive assays can also be utilized that employ a similar method, but the target cells are pre-loaded with fluorescent dyes, such as calcein-AM, CFSE, BCECF, or lanthanide flurophore (Europium). See, e.g., Parekh, Bhavin S et al. “Development and validation of an antibody-dependent cell-mediated cytotoxicity- reporter gene assay.” mAbs vol. 4,3 (2012): 310-8. Doi:10.4161 / mabs.19873, the entire contents of which is incorporated by reference herein for all purposes. Exemplary commercially available non-radioactive assays include, for example, ACTI™ non-radioactive cytotoxicity assay for flow cytometry (Cell Technology, Inc. Mountain View, Calif.; and CytoTox 96® non-radioactive cytotoxicity assay (Promega, Madison, Wis.). Additional non-limiting examples of in vitro assays that can be used to assess ADCC activity of a protein (e.g., antibody) described herein include those described in US5500362; US5821337; Hellstrom, I., et al., Proc. Nat’l Acad. Sci. USA 83 (1986) 7059-7063; Hellstrom, I., et al., Proc. Nat’l Acad. Sci. USA 82 (1985) 1499-1502; and Bruggemann, M., et al., J. Exp. Med. 166 (1987) 1351-1361, the entire contents of each of which is incorporated by reference herein. Alternatively, or additionally, ADCC activity of a protein (e.g., an antibody) described herein may be assessed in vivo, e.g., in an animal model such as that disclosed in Clynes, et al., Proc. Nat’l Acad. Sci. USA 95 (1998) 652-656, the entire contents of which is incorporated by reference herein for all purposes.

[0269] C1q binding assays can be utilized to assess the ability of a an antibody described herein to bind C1q (or bind with less affinity than a reference protein) and hence lack (or have decreased) CDC activity. The binding of an antibody described herein to C1q can be determined by a variety of in vitro assays (e.g., biochemical or immunological based assays) known in the art for determining Fc-C1q interactions, including e.g., equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)), or kinetic methods (e.g., surface plasmon resonance (SPR) analysis), and other methods such as indirect binding assays,Attorney Docket No.63340.10WO01 competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). These and other methods may utilize a label on one or more of the components being examined and / or employ a variety of detection methods including but not limited to chromogenic, fluorescent, luminescent, or isotopic labels. A detailed description of binding affinities and kinetics can be found in e.g., Paul, W. E., ed., Fundamental Immunology, 4thEd., Lippincott-Raven, Philadelphia (1999), the entire contents of which is incorporated by reference herein. For example, see, e.g., C1q and C3c binding ELISAs described in WO2006 / 029879 and WO2005 / 100402, the entire contents of each of which is incorporated by reference herein for all purposes. Additional CDC activity assays include those described in e.g., Gazzano-Santoro, et al., J. Immunol. Methods 202 (1996) 163; Cragg, M. S., et al., Blood 101 (2003) 1045-1052; and Cragg, M. S., and Glennie, M. J., Blood 103 (2004) 2738-2743), the entire contents of each of which is incorporated by reference herein for all purposes.

[0270] ADCP activity can be measured by in vitro or in vivo methods known in the art and also commercially available assays (see, e.g., van de Donk NW, Moreau P, Plesner T, et al. “Clinical efficacy and management of monoclonal antibodies targeting CD38 and SLAMF7 in multiple myeloma,” Blood, 127(6):681‐695 (2016), the entire contents of each of which is incorporated by reference herein for all purposes). For example, a primary cell based ADCP assay can be used in which fresh human peripheral blood mononuclear cells (PBMCs) are isolated, monocytes isolated and differentiated in culture to macrophages using standard procedures. The macrophages are fluorescently labeled added to cultures containing fluorescently labeled target cells. Phagocytosis events can be analyzed using FACS screening and / or microscopy. A modified reporter version of the above described assay can also be used that employs an engineered cell line that stably expresses FcγRIIa (CD32a) as the effector cell line (e.g., an engineered T cell line, e.g., THP-1), removing the requirement for primary cells. Exemplary ADCP assays are described in e.g., Ackerman, M. E. et al. A robust, high-throughput assay to determine the phagocytic activity of clinical antibody samples. J. Immunol. Methods 366, 8–19 (2011); and Mcandrew, E. G. et al. Determining the phagocytic activity of clinical antibody samples. J. Vis. Exp. 3588 (2011). Doi:10.3791 / 3588; the entire contents of each of which is incorporated by reference herein.

[0271] Binding of an antibody described herein to an Fc receptor can be determined by a variety of in vitro assays (e.g., biochemical or immunological based assays) known in the art for determining Fc-Fc receptor interactions, i.e., specific binding of an Fc region to an Fc receptor.Attorney Docket No.63340.10WO01 Common assays include equilibrium methods (e.g., enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA)), or kinetic methods (e.g., surface plasmon resonance (SPR) analysis), and other methods such as indirect binding assays, competitive inhibition assays, fluorescence resonance energy transfer (FRET), gel electrophoresis, and chromatography (e.g., gel filtration). These and other methods may utilize a label on one or more of the components being examined and / or employ a variety of detection methods including but not limited to chromogenic, fluorescent, luminescent, or isotopic labels. A detailed description of binding affinities and kinetics can be found in e.g., Paul, W. E., ed., Fundamental Immunology, 4” Ed., Lippincott-Raven, Philadelphia (1999), the entire contents of which is incorporated by reference herein for all purposes.

[0272] In some embodiments, the Fc region of an antibody described herein is varied (e.g., comprises one or more amino acid variation (e.g., one or more amino acid substitution, deletion, addition, etc.)) (referred to herein as a “varied Fc region”), relative to the amino acid sequence of a reference Fc region (e.g., a wild type Fc region). In some embodiments, the one or more amino acid variation (e.g., the one or more amino acid substitution, deletion, addition, etc.)) decreases or abolishes one or more Fc effector function, relative to a reference Fc that does not comprise the variation (e.g., the one or more variation (e.g., the one or more amino acid substitution, deletion, addition, etc.)).

[0273] In some embodiments, the antibody comprising a varied Fc region exhibits no detectable or decreased ADCC compared to a reference antibody that does not comprise the Fc region variation (e.g., the one or more amino acid variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc region exhibits no detectable or decreased CDC compared to a reference antibody that does not comprise the Fc region variation (e.g., the one or more amino acid variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a modified Fc region exhibits no detectable or decreased ADCP compared to a reference antibody that does not comprise the Fc region variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc region exhibits decreased or no detectable specific binding affinity to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))) compared to a reference antibody that does not comprise the FcAttorney Docket No.63340.10WO01 region variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc region exhibits decreased or no detectable specific binding affinity to FcγRI, FcγIIa, and / or FcγIIIa compared to an antibody that does not comprise the Fc region variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc exhibits decreased or no detectable specific binding affinity to FcγRI compared to a reference antibody that does not comprise the Fc variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc exhibits decreased or no detectable specific binding affinity to FcγIIa compared to a reference antibody that does not comprise the Fc region variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc region exhibits decreased or no detectable specific binding affinity to FcγIIIa compared to an antibody that does not comprise the Fc region variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)). In some embodiments, the antibody comprising a varied Fc region exhibits decreased or no detectable specific binding affinity to C1q compared to a reference antibody that does not comprise the Fc region variation (e.g., the one or more variation (e.g., one or more amino acid substitution, deletion, or addition)).

[0274] In some embodiments, the antibody comprising an Fc region exhibits no detectable ADCC. In some embodiments, the antibody comprising an Fc region exhibits no detectable CDC. In some embodiments, the antibody comprising an Fc region exhibits no detectable ADCP. In some embodiments, the antibody comprising an Fc region exhibits no detectable specific binding affinity to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))). In some embodiments, the antibody comprising an Fc region exhibits no detectable specific binding affinity to FcγRI, FcγIIa, and / or. In some embodiments, the antibody comprising an Fc exhibits no detectable specific binding affinity to FcγRI. In some embodiments, the antibody comprising an Fc exhibits no detectable specific binding affinity to FcγIIa. In some embodiments, the antibody comprising an Fc region exhibits no detectable specific binding affinity to FcγIIIa. In some embodiments, the antibody comprising an Fc region exhibits no detectable specific binding affinity to C1q.

[0275] Amino acid substitutions that decrease or abolish one or more Fc effector function areAttorney Docket No.63340.10WO01 known in the art. See for example, Saunders Kevin, “Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life,” Frontiers in Immunology, v10 (June 7, 2019) DOI=10.3389 / fimmu.2019.01296, the full contents of which is incorporated by reference herein for all purposes, see more particularly for example, e.g., Table 4 of Saunders.

[0001] Table 17 below, provides exemplary amino acid substitutions (and combinations thereof) that can be utilized to increase one or more Fc effector function. Amino acids in Table 17 are numbered according to the EU numbering scheme. The effects on effector function set forth in Table 17 are exemplary only and not intended to be limiting. The amino acid substitutions set forth in Table 17 (except where noted) are with reference to an IgG1 Fc region. However, a person of ordinary skill in the could identify the corresponding amino acid in a non-IgG1 Fc region, for example in an IgG2 or IgG4 Fc region, should the base amino acid be different between the IgG1 and non-IgG1 Fc region. Table 17. Exemplary Ig Fc Variations and Glycoengineering to Decrease Effector Function. Change Exemplary Effects on Effector FunctionAttorney Docket No.63340.10WO01 IgG2 – H268Q / V309L / A330S / P331S Reduces ADCC, ADCP, CDC Glycoengineering neor more of the amino acid substitutions set forth in Table 17 (set forth in any set). In some embodiments, the hIg Fc (e.g., IgG1 Fc) region comprises any one or more of the sets of amino acid substitutions set forth in Table 17. In some embodiments, the hIg Fc (e.g., IgG1 Fc) region comprises any one or more of the glycosylation changes set forth in Table 17.

[0003] In some embodiments, the hIg Fc (e.g., IgG1 Fc) region comprises an amino acid substitution at any one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more) of amino acid positions L234, L235, P329, P331, D265, G237, E318, E233, G236, L328, D270, K322, V264, F241, and / or N297. In some embodiments, the hIg Fc (e.g., IgG1 Fc) region comprises an amino acid substitution at from about 1-10 (e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) of the following amino acid positions L234, L235, P329, P331, D265, G237, E318, E233, G236, L328, D270, K322, V264, F241, and / or N297.

[0004] In some embodiments, the hIg Fc (e.g., IgG1 Fc) region comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more) of the following amino acid substitutions L234A, L234G, L235F, L235E, L235A, P329G, P329A, P331S, D265A, G237A, E318A, E233P, G236R, L328R, D270A, K322A, V264A, F241A, N297A, N297G, and / or N297Q. In some embodiments, the hIg Fc (e.g., IgG1 Fc) region comprises from about 1-10 (e.g., 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, or 1-2) of the following amino acid substitutions L234A, L234G, L235F, L235E, L235A, P329G, P329A, P331S, D265A, G237A, E318A, E233P, G236R, L328R, D270A, K322A, V264A, F241A, N297A, N297G, or N297Q.

[0005] In some embodiments, the hIg Fc region comprises a hIgG1 Fc region comprising one or more amino acid variation.

[0006] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position L234, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises and L234A or L234G amino acid substitution, EU numbering according to Kabat.

[0007] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position L235, EU numbering according to Kabat. In some embodiments, the hIgG1Attorney Docket No.63340.10WO01 Fc region comprises and L235A, L235G, L235E, or L235F amino acid substitution, EU numbering according to Kabat.

[0008] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position P329, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises and P329A or P329G amino acid substitution, EU numbering according to Kabat.

[0009] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions L234 and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions L234A and / or L235A, EU numbering according to Kabat.

[0010] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions L234, L235, and / or P329, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions L234A, L235A, and / or P329G, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions L234A, L235A, and / or P329A, EU numbering according to Kabat.

[0011] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions P331, L234, and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions P331S, L234G, and / or L235F, EU numbering according to Kabat.

[0012] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position D265, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a D235A amino acid substitution, EU numbering according to Kabat.

[0013] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position G237, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a G237A amino acid substitution, EU numbering according to Kabat.

[0014] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position E318, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a E318A amino acid substitution, EU numbering according to Kabat.Attorney Docket No.63340.10WO01

[0015] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position E233, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a E233P amino acid substitution, EU numbering according to Kabat.

[0016] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position D270, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a D270A amino acid substitution, EU numbering according to Kabat.

[0017] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position K322, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a K322A amino acid substitution, EU numbering according to Kabat.

[0018] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position P331, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a P331A amino acid substitution, EU numbering according to Kabat.

[0019] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position F241, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a F241A amino acid substitution, EU numbering according to Kabat.

[0020] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid position N297, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises a N297A, N297G, or N297Q amino acid substitution, EU numbering according to Kabat.

[0021] In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions G236 and / or L328, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions G236R and / or L328R, EU numbering according to Kabat.

[0022] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at amino acid position S228, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises an S228P amino acid substitution, EU numbering according to Kabat.

[0023] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at amino acid position F234, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises an F234A amino acid substitution, EU numbering according to Kabat.

[0024] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at amino acid position L235A, EU numbering according to Kabat. In some embodiments, the hIgG1Attorney Docket No.63340.10WO01 Fc region comprises an L235A, L235G, or L235E amino acid substitution, EU numbering according to Kabat.

[0025] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at one or more (e.g., 1 or 2) of amino acid positions S228 and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S228P and / or L235E, EU numbering according to Kabat.

[0026] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S228P, F234A, and / or L235A, EU numbering according to Kabat.

[0027] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S228P, F234A, and / or L235G, EU numbering according to Kabat.

[0028] In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, or 3) of amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc region comprises one or more (e.g., 1, 2, or 3) of the following amino acid substitutions S228P, F234A, and / or L235E, EU numbering according to Kabat.

[0029] In some embodiments, the hIgG2 Fc region comprises an amino acid substitution at one or more (e.g., 1, 2, 3, or 4) of amino acid positions H268, V309, A330, and / or P331, EU numbering according to Kabat. In some embodiments, the hIgG2 Fc region comprises one or more (e.g., 1, 2, 3, or 4) of the following amino acid substitutions H268Q, V309L, A330S, and / or P331S, EU numbering according to Kabat.

[0030] In some embodiments, the hIg Fc region of a CLEC2D binding protein (or a conjugate or fusion protein comprising the same) (e.g., described herein) comprises one or more changes to the glycosylation. In some embodiments, the hIg Fc region of a CLEC2D binding protein (or a conjugate or fusion protein comprising the same) (e.g., described herein) has increased high mannose glycosylation.Attorney Docket No.63340.10WO01

[0031] In some embodiments, the varied Fc comprises a hIgG1 Fc region comprising one or more amino acid variations (e.g., one or more amino acid substitutions). In some embodiments, the hIgG1 Fc region comprises an amino acid substitution at amino acid positions L234, L235, and / or P329, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises the following amino acid substitutions L234A and / or L235A, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises the following amino acid substitutions L234A, L235A, and P329G, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises the following amino acid substitutions L234A, L235A, and P329A, EU numbering according to Kabat. In some embodiments, the hIgG1 Fc region comprises the following amino acid substitutions L234A, L235A, and P329S, EU numbering according to Kabat.

[0032] In some embodiments, the varied Fc region comprises a hIg4 Fc region comprising one or more amino acid variations (e.g., one or more amino acid substitutions). In some embodiments, the hIgG4 Fc region comprises an amino acid substitution at amino acid positions S228, F234, and / or L235, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc region comprises the following amino acid substitutions S228P, F234A, and / or L235A, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc region comprises the following amino acid substitutions S228P, F234A, and / or L235E, EU numbering according to Kabat. In some embodiments, the hIgG4 Fc comprises the following amino acid substitutions S228P and / or L235E, EU numbering according to Kabat.

[0033] The amino acid sequence of exemplary varied Fc regions that are known in the art to exhibit a decrease in one more effector function is provided in Table 4. Table 4. The Amino Acid Sequence of Exemplary Varied Fc Regions. SEQ D i ti A i A id S IDAttorney Docket No.63340.10WO01 KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS Without C-terminal FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG L sineAttorney Docket No.63340.10WO01 hIgG1 Partial Hinge TCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH Region + CH2 Region + EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD CH3 Re ion WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDAttorney Docket No.63340.10WO01 Without C-terminal L sineAttorney Docket No.63340.10WO01 hIgG1 Hinge Region + EPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEV CH2 Region + CH3 TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV Re ion SVLTVLHQDWLNGKEYKCKVSNKALSAPIEKTISKAKGQPREPQAttorney Docket No.63340.10WO01 hIgG4 Hinge Region + ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCV CH2 Region + CH3 VVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL Re ion TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTing an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 4.

[0035] In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises or consists of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., substitutions, additions, deletions, etc.). In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises or consists of about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., substitutions, additions, deletions, etc.). In someAttorney Docket No.63340.10WO01 embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises or consists of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., substitutions, additions, deletions, etc.).

[0036] In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid substitutions. In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises or consists of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises or consists of about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of a polypeptide set forth in Table 4, and further comprises or consists of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions.

[0037] In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89.

[0038] In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid variations (e.g., amino acid substitutions, deletions, or additions). In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises or consists of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., substitutions, additions, deletions, etc.). In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises or consists of about 1, 2, 3, 4,Attorney Docket No.63340.10WO01 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., substitutions, additions, deletions, etc.). In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises or consists of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid variations (e.g., substitutions, additions, deletions, etc.).

[0039] In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises 1 or more but less than 15% (less than 12%, less than 10%, less than 8%), amino acid substitutions. In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises or consists of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises or consists of about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions. In some embodiments, the amino acid sequence of the variant hIg antibody comprises a hIg Fc region that comprises or consists of the amino acid sequence of any one of SEQ ID NOS: 58-89, and further comprises or consists of no more than about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions.

[0040] In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and an alanine at position L235, EU numbering according to Kabat. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises of an alanine, glycine, or serine at position P329, EU numbering according to Kabat.

[0041] In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234; an alanine at position L235; and an alanine, glycine, or serine at position P329 EU numbering according to Kabat. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises of an alanine at position L234; an alanine at position L235; and an alanine at position P329 EU numbering according to Kabat. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234; an alanine at position L235; and a glycine amino acid at position P329 EU numbering according to Kabat. In someAttorney Docket No.63340.10WO01 embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234; an alanine at position L235; and a serine amino acid at position P329 EU numbering according to Kabat.

[0042] In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and / or an alanine, glycine, or serine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and an alanine, glycine, or serine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and an alanine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and a glycine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at positionAttorney Docket No.63340.10WO01 L235, and a serine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4.

[0043] In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and / or an alanine, glycine, or serine amino acid at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58- 89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and an alanine, glycine, or serine amino acid at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and an alanine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and a glycine amino acid at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In someAttorney Docket No.63340.10WO01 embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and a serine amino acid at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89.

[0044] In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and / or an alanine, glycine, or serine amino acid at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58- 89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234 and an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and an alanine, glycine, or serine amino acid at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and an alanine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and a glycine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identicalAttorney Docket No.63340.10WO01 to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG1 Fc region comprises an alanine at position L234, an alanine at position L235, and a serine at position P329, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89.

[0045] In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234 and an alanine at position L235, EU numbering according to Kabat. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a proline at position S228, EU numbering according to Kabat.

[0046] In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234; an alanine at position L235; and a proline at position S228, EU numbering according to Kabat.

[0047] In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234, an alanine at position L235, and / or a proline at position S228, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234, an alanine at position L235, and a proline at position S228, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a polypeptide set forth in Table 3 or 4.

[0048] In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234, an alanine at position L235, and / or a proline at position S228, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%,Attorney Docket No.63340.10WO01 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS:199-224. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234, an alanine at position L235, and a proline at position S228, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89.

[0049] In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234, an alanine at position L235, and / or a proline at position S228, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234 and / or an alanine at position L235, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. In some embodiments, the amino acid sequence of the hIgG4 Fc region comprises a phenylalanine at position L234, an alanine at position L235, and a proline at position S228, EU numbering according to Kabat; and comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 58-89. 5.3.3.2 Promotion of Heterodimerization

[0050] As described herein, in some embodiments, the antibody comprises a first and second Fc region (see, e.g., § 5.3.3). In some embodiments, the first Ig Fc region and the second Ig Fc region each comprise one or more amino acid modifications relative to each other to promote heterodimerization. IgG derived heterodimeric formats can be generated by methods known in the art, e.g., by forced heavy chain heterodimerization. Forced heavy chain heterodimerization can beAttorney Docket No.63340.10WO01 obtained using known methods in the art, e.g., knob-in-hole or strand exchange engineered domains (SEED), see, e.g., Ji-Hee et al., “Immunoglobulin Fc Heterodimer Platform Technology: From Design to Applications in Therapeutic Antibodies and Proteins” Frontiers in Immunology, v7(article 394) (2016) DOI=10.3389 / fimmu.2016.00394 (hereinafter “Ji-Hee 2016”), the entire contents of which is incorporated by reference herein for all purposes.

[0051] In some embodiments, an interface of the first and the second Ig Fc regions is varied, e.g., introduction of an amino acid substitution, to increase heterodimerization, e.g., relative to a non-modified interface, e.g., a naturally occurring interface. For example, dimerization of the first and second Ig Fc regions can be enhanced by providing an Ig Fc interface of a first and a second Fc region with one or more of: a paired protuberance-cavity (“knob-in-hole”), an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms, e.g., relative to a non-modified interface.

[0052] Knob-in-Hole amino acid pairing modifications are known in the art, and described in e.g., US5731116; US7476724; Ji-Hee 2016; and Ridgway, J. “'Knobs-into-holes' engineering of antibody CH3 domains for heavy chain heterodimerization” et al. Prot. Engineering 9(7): 617-621 (1996), the full contents of each of which is incorporated by reference herein. Generally, Knob-in- Hole comprises 1) introducing one or more amino acid substitutions in the CH3 domain of one or both of the first and second subject Ig Fc regions to promote heterodimerization; and 2) combining the modified Ig Fc regions under conditions that promote heterodimerization. “Knobs” are typically created by substituting a small amino acid in a parental Ig Fc region with a larger amino acid (e.g., T366Y or T366W); “holes” are created by substituting a larger residue in a parental Ig Fc region with a smaller amino acid (e.g., Y407T, T366S, 11368A, or Y407V). Exemplary Knob- in-Hole mutations include S354C, T366W in the “knob” Ig Fc region and Y349C, T366S, L368A, Y407V in the “hole” Ig Fc region. Other exemplary Knob-in-Hole mutations, which can be incorporated into any one or more of the embodiments, are provided in Table 5, with additional exemplary optional stabilizing Ig Fc cysteine mutations. Table 5. Exemplary Knob-in-hole and Stabilizing Cysteine Modifications. Amino Acid Position Knob Ig Fc Amino Acid Hole Ig Fc Amino AcidAttorney Docket No.63340.10WO01 Y407 - Y407V Stabilizing Cysteine Amino Acid Substitutions S354 S354C teheterodimerization is provided in Table 6. Table 6. The Amino Acid Sequence of Exemplary Pairs of Varied Heterodimeric Fc Regions. Description Amino Acid SequenceSEQID NOAttorney Docket No.63340.10WO01 hIgG1 Hinge Region + EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE CH2 Region + CH3 VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR Re ion VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRAttorney Docket No.63340.10WO01 Without C-terminal Lysine hIG1 Hin R in + EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEAttorney Docket No.63340.10WO01 hIgG1 Partial Hinge TCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS Region + CH2 Region + HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH CH3 Re ion QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPAttorney Docket No.63340.10WO01 Without C-terminal Lysine hIG1 CH2 R in + PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDAttorney Docket No.63340.10WO01 EPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQP Hole ENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHE T366S / L368A / Y407V / Y ALHNHYTQKSLSLSPGion and a second Ig Fc region.

[0055] In some embodiments, the amino acid sequence of the first Fc region comprises a T366W amino acid substitution, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, and Y407V, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3). In some embodiments, the amino acid sequence of the first hIg further comprises a S354C amino acid substitution, EU numbering according to Kabat; and the amino acid sequence of the second Fc region comprises a Y349C amino acid substitution, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0056] In some embodiments, the amino acid sequence of the first Fc region comprises each of the following amino acid substitutions: T366W and a S354C, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the followingAttorney Docket No.63340.10WO01 amino acid substitutions: T366S, L368A, Y407V, and Y349C, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0057] In some embodiments, the amino acid sequence of the second Fc region comprises a T366W amino acid substitution, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, and Y407V, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3). In some embodiments, the amino acid sequence of the second hIg further comprises a S354C amino acid substitution, EU numbering according to Kabat; and the amino acid sequence of the second Fc region comprises a Y349C amino acid substitution, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0058] In some embodiments, the amino acid sequence of the second Fc region comprises each of the following amino acid substitutions: T366W and a S354C, EU numbering according to Kabat; and the second the amino acid sequence of the Fc region comprises each of the following amino acid substitutions: T366S, L368A, Y407V, and Y349C, EU numbering according to Kabat; each relative to the amino acid sequence of an exemplary reference Ig Fc region (e.g., a reference Ig Fc region set forth in Table 3).

[0059] In some embodiments, the amino acid sequence of the first Ig Fc region comprises a W amino acid at position T366, EU numbering according to Kabat; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat.

[0060] In some embodiments, the amino acid sequence of the first Ig Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat.

[0061] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to theAttorney Docket No.63340.10WO01 amino acid sequence of any one of SEQ ID NOS: 90-97; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 106-113.

[0062] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 98-105; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NOS: 114-121.

[0063] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 90; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 106.

[0064] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 91; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 107.

[0065] In some embodiments, the amino acid sequence of the first Fc region comprises a WAttorney Docket No.63340.10WO01 amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 92; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 108.

[0066] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 93; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 109.

[0067] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 94; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 110.

[0068] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 95; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 111.Attorney Docket No.63340.10WO01

[0069] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 96; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 112.

[0070] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 97; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, and a V amino acid at position Y407, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 113.

[0071] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 98; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 114.

[0072] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 99; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at positionAttorney Docket No.63340.10WO01 Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 115.

[0073] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 100; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 116.

[0074] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 101; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 117.

[0075] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 102; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 118.

[0076] In some embodiments, the amino acid sequence of the first Fc region comprises a WAttorney Docket No.63340.10WO01 amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 103; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 119.

[0077] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 104; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 120.

[0078] In some embodiments, the amino acid sequence of the first Fc region comprises a W amino acid at position T366 and a C amino acid at position S354, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 105; and the amino acid sequence of the second Ig Fc region comprises a S amino acid at position T366, an A amino acid at position L368, a V amino acid at position Y407, and a C amino acid at position Y349, EU numbering according to Kabat, and is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of any one of SEQ ID NO: 121. 5.3.3.3 Ig Constant Region Variations and Manipulations for Site Specific Conjugation

[0079] In some embodiments, the molecular payload (e.g., CpG ODN) is conjugated (e.g., directly, or indirectly through a linker) to the Ig constant region. In some embodiments, the molecular payload (e.g., CpG ODN) (or a linker) is conjugated directly to an amino acid (e.g., aAttorney Docket No.63340.10WO01 naturally occurring amino acid or an engineered (i.e., variant) amino acid) within the Ig constant region.

[0080] In some embodiments, the molecular payload (e.g., CpG ODN) (or the linker) is conjugated directly to an engineered lysine, cysteine, or tyrosine amino acid residue within the Ig constant region. In some embodiments, the amino acid sequence of the Ig constant region comprises the substitution of one or more naturally occurring amino acid residue with a lysine, cysteine, or tyrosine amino acid residue (e.g., to mediate conjugation). In some embodiments, the amino acid sequence of the Ig constant region comprises the substitution of one or more non- cysteine amino acid residue with a cysteine amino acid residue (e.g., to mediate conjugation). In some embodiments, the amino acid sequence of the Ig constant region comprises the substitution of one or more non-lysine amino acid residue with a lysine amino acid residue (e.g., to mediate conjugation). In some embodiments, the amino acid sequence of the Ig constant region comprises the substitution of one or more non-tyrosine amino acid residue with a tyrosine amino acid residue (e.g., to mediate conjugation).

[0081] In some embodiments, the amino acid sequence of the Ig constant region comprises the addition of one or more lysine, cysteine, or tyrosine amino acid residue (e.g., to mediate conjugation). In some embodiments, the amino acid sequence of the Ig constant region comprises the addition of one or more lysine amino acid residue (e.g., to mediate conjugation). In some embodiments, the amino acid sequence of the Ig constant region comprises the addition of one or more cysteine amino acid residue (e.g., to mediate conjugation). In some embodiments, the amino acid sequence of the Ig constant region comprises the addition of one or more tyrosine amino acid residue (e.g., to mediate conjugation).

[0082] In some embodiments, the amino acid sequence of the Ig constant region comprises a cysteine at position S440, EU numbering according to Kabat. In some embodiments, the amino acid sequence of the Ig constant region comprises a cysteine at position S442, EU numbering according to Kabat. See, e.g., US8834885B2 and WO1996014339, the entire contents of each of which are incorporated herein by reference.

[0083] In some embodiments, the glycosylation of the antibody is manipulated (e.g., deglycosylated) prior to conjugation. Standard suitable methods of deglycosylation are known, e.g., PNGase F treatment. See, e.g., Benjamin SR, Jackson CP, Fang S, Carlson DP, Guo Z, Tumey LN. Thiolation of Q295: Site-Specific Conjugation of Hydrophobic Payloads without the Need forAttorney Docket No.63340.10WO01 Genetic Engineering. Mol Pharm. 2019 Jun 3;16(6):2795-2807. doi: 10.1021 / acs.molpharmaceut.9b00323. Epub 2019 May 17. PMID: 31067063.; Park, J., Chariou, P. L., & Steinmetz, N. F. (2020). Site-Specific Antibody Conjugation Strategy to Functionalize Virus-Based Nanoparticles. Bioconjugate chemistry, 31(5), 1408–1416. https: / / doi.org / 10.1021 / acs.bioconjchem.0c00118, the entire contents of each of which is incorporated herein by reference for all purposes. In some embodiments, the antibody is deglycosylated in the absence of the introduction of an amino acid variation to mediate conjugation. 5.3.4 Methods of Making Proteins (e.g., Targeting Agents)

[0084] Any of the proteins (e.g., antibodies (e.g., anti-CLEC2D antibodies)), including the targeting moieties described herein, may be produced using standard methods known in the art. For example, each may be produced by recombinant technology in host cells (e.g., insect cells, mammalian cells, bacteria) that have been transfected or transduced with a nucleic acid expression vector (e.g., plasmid, viral vector (e.g., a baculoviral expression vector)) encoding the protein (e.g., antibody). Such general methods are common knowledge in the art. The expression vector typically contains an expression cassette that includes nucleic acid sequences capable of bringing about expression of the nucleic acid molecule encoding the protein interest, such as promoter(s), enhancer(s), polyadenylation signals, and the like. The person of ordinary skill in the art is aware that various promoter and enhancer elements can be used to obtain expression of a nucleic acid molecule in a host cell. For example, promoters can be constitutive or regulated, and can be obtained from various sources, e.g., viruses, prokaryotic or eukaryotic sources, or artificially designed. Post transfection or transduction, host cells containing the expression vector encoding the protein of interest are cultured under conditions conducive to expression of the nucleic acid molecule encoding the protein. Culture media is available from various vendors, and a suitable medium can be routinely chosen for a host cell to express a protein or polypeptide of interest. Host cells can be adherent or suspension cultures, and a person of ordinary skill in the art can optimize culture methods for specific host cells selected. For example, suspension cells can be cultured in, for example, bioreactors in e.g., a batch process or a fed-batch process. The produced protein or polypeptide may be isolated from the cell cultures, by, for example, column chromatography in either flow-flow through or bind-and-elute modes. Examples include, but are not limited to, ionAttorney Docket No.63340.10WO01 exchange resins and affinity resins, such as lentil lectin Sepharose, and mixed mode cation exchange-hydrophobic interaction columns (CEX-HIC). The protein or polypeptide may be concentrated, buffer exchanged by ultrafiltration, and the retentate from the ultrafiltration may be filtered through an appropriate filter, e.g., a 0.22µm filter. See, e.g., Hacker, David (Ed.), Recombinant Protein Expression in Mammalian Cells: Methods and Protocols (Methods in Molecular Biology), Humana Press (2018); and McPherson et al., “Development of a SARS Coronavirus Vaccine from Recombinant Spike Protein Plus Delta Inulin Adjuvant,” Chapter 4, in Sunil Thomas (ed.), Vaccine Design: Methods and Protocols: Volume 1: Vaccines for Human Diseases, Methods in Molecular Biology, Springer, New York, 2016. See also U.S. Pat.5,762,939, the entire contents of each of which is incorporated by reference herein for all purposes.

[0085] The proteins (e.g., antibodies), described herein may also be produced synthetically. 5.4 CpG ODNs

[0086] As described above, the conjugates described herein comprise at least one CpG ODN. As described herein, CpG ODNs comprise one or more CpG motif and are capable of activating toll like receptor (TLR) 9. TLR9 is a member of the toll like receptor family that recognize conserved patterns on microorganisms known as pathogen associated molecular patterns (PAMPs). TLR9 is expressed primarily on antigen presenting cells such as B cells and dendritic cells (e.g., plasmacytoid dendritic cells) (e.g., in the endosome and / or lysosome). The amino acid sequence of an exemplary reference mature human TLR9 (hTLR9) protein is set forth in Table 7 below. Table 7. Amino Acid Sequence of Reference hTLR9. Description Amino Acid SequenceSEQOAttorney Docket No.63340.10WO01 WAEGDLYLHFFQGLSGLIWLDLSQNRLHTLLPQTLRNLPKSLQVLRLRD NYLAFFKWWSLHFLPKLEVLDLAGNQLKALTNGSLPAGTRLRRLDVSCN SISFVAPGFFSKAKELRELNLSANALKTVDHSWFGPLASALQILDVSANfly, as described above, Class A CpG ODNs comprise a natural phosphodiester (PO) backbone central CpG-containing palindromic motif and a phosphorothioated (PS) modified 3' poly-G string. Class B CpG ODNs comprise a full phosphorothioated (PS) modified backbone. Class C CpG ODNs comprise a full phosphorothioated (PS) modified backbone and a CpG-containing palindromic motif. In some embodiments, the CpG ODN is a class A CpG ODN. In some embodiments, the CpG ODN is a class B CpG ODN. In some embodiments, the CpG ODN is a class C CpG ODN. In some embodiments, the CpG ODN comprises is single stranded. In some embodiments, the CpG ODN is double stranded. In some embodiments, the CpG ODN comprises only DNA nucleotides. 5.4.1 Overall Length

[0088] In some embodiments, the CpG ODN comprises from about 10-100, 10-90, 10-80, 10- 70, 10-60, 10-50, 10-40, 10-30, 10-20, 20-100, 20-90, 20-80, 20-70, 20-60, 20-50, 20-40, 20-30, 30-100, 30-90, 30-80, 30-70, 30-60, 30-50, 30-40, 40-100, 40-90, 40-80, 40-70, 40-60, 40-50, 50- 100, 50-90, 50-80, 50-70, 50-60, 60-600, 60-90, 60-80, 60-70, 70-700, 70-90, 70-80, 80-800, 80- 90, or 90-100.

[0089] In some embodiments, the CpG ODN comprises from about 10-30, 10-29, 10-28, 10- 27, 10-26, 10-25, 10-24, 10-23, 10-22, 10-21, 10-20, 10-19, 10-18, 10-17, 10-16, 10-15, 10-14, 10-13, 10-12, 10-11, 11-30, 11-29, 11-28, 11-27, 11-26, 11-25, 11-24, 11-23, 11-22, 11-21, 11-20, 11-19, 11-18, 11-17, 11-16, 11-15, 11-14, 11-13, 11-12, 12-30, 12-29, 12-28, 12-27, 12-26, 12-25, 12-24, 12-23, 12-22, 12-21, 12-20, 12-19, 12-18, 12-17, 12-16, 12-15, 12-14, 12-13, 13-30, 13-29, 13-28, 13-27, 13-26, 13-25, 13-24, 13-23, 13-22, 13-21, 13-20, 13-19, 13-18, 13-17, 13-16, 13-15, 13-14, 14-30, 14-29, 14-28, 14-27, 14-26, 14-25, 14-24, 14-23, 14-22, 14-21, 14-20, 14-19, 14-18, 14-17, 14-16, 14-15, 15-30, 15-29, 15-28, 15-27, 15-26, 15-25, 15-24, 15-23, 15-22, 15-21, 15-20,Attorney Docket No.63340.10WO01 15-19, 15-18, 15-17, 15-16, 16-30, 16-29, 16-28, 16-27, 16-26, 16-25, 16-24, 16-23, 16-22, 16-21, 16-20, 16-19, 16-18, 16-17, 17-30, 17-29, 17-28, 17-27, 17-26, 17-25, 17-24, 17-23, 17-22, 17-21, 17-20, 17-19, 17-18, 18-30, 18-29, 18-28, 18-27, 18-26, 18-25, 18-24, 18-23, 18-22, 18-21, 18-20, 18-19, 19-30, 19-29, 19-28, 19-27, 19-26, 19-25, 19-24, 19-23, 19-22, 19-21, 19-20, 20-30, 20-29, 20-28, 20-27, 20-26, 20-25, 20-24, 20-23, 20-22, 20-21, 21-30, 21-29, 21-28, 21-27, 21-26, 21-25, 21-24, 21-23, 21-22, 22-30, 22-29, 22-28, 22-27, 22-26, 22-25, 22-24, 22-23, 23-30, 23-29, 23-28, 23-27, 23-26, 23-25, 23-24, 24-30, 24-29, 24-28, 24-27, 24-26, 24-25, 25-30, 25-29, 25-28, 25-27, 25-26, 26-30, 26-29, 26-28, 26-27, 27-30, 27-29, 27-28, 28-30, 28-29, or 29-30.

[0090] In some embodiments, the CpG ODN comprises from about 15-30, 15-25, 15-20, 16- 30, 16-25, 16-20, 17-30, 17-25, 17-20, 18-30, 18-25, 18-20, 19-30, 19-25, or 19-20 nucleotides.

[0091] In some embodiments, the CpG ODN comprises or consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, or 41 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 15 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 16 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 17 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 18 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 19 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 20 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 21 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 22 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 23 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 24 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 25 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 26 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 27 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 28 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 29 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 30 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 31 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 32 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 33 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 34 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 35 nucleotides. In some embodiments, the CpG ODN comprises or consists ofAttorney Docket No.63340.10WO01 about 36 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 37 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 38 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 39 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 40 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 41 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 42 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 43 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 44 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 45 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 46 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 47 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 48 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 49 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 50 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 51 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 52 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 53 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 54 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 55 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 56 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 57 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 58 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 59 nucleotides. In some embodiments, the CpG ODN comprises or consists of about 60 nucleotides. 5.4.2 CpG Motif Content

[0092] In some embodiments, the CpG ODN comprises at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more CpG motifs. In some embodiments, the CpG ODN comprises from about 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 13, or 1-2 CpG motifs. In some embodiments, the CpG ODN comprises about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more CpG motifs.

[0093] In some embodiments, the CpG ODN consists of at least about 1, 2, 3, 4, 5, 6, 7, 8, 9,Attorney Docket No.63340.10WO01 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more CpG motifs. In some embodiments, the CpG ODN consists of from about 1-30, 1-29, 1-28, 1-27, 1-26, 1-25, 1-24, 1-23, 1-22, 1-21, 1-20, 1-19, 1-18, 1-17, 1-16, 1-15, 1-14, 1-13, 1-12, 1-11, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 13, or 1-2 CpG motifs. In some embodimen...

Claims

Attorney Docket No.63340.10WO01 CLAIMS What is claimed is:

1. A conjugate comprising: (a) an antibody that specifically binds to C-type lectin domain family 2 member D (CLEC2D) (e.g., human CLEC2D (hCLEC2D)); and (b) at least one CpG oligodeoxynucleotide (CpG ODN).

2. The conjugate of claim 1, wherein the antibody comprises or consists of a full-length antibody, a Fab, a Fab', a F(ab')2, a Fab-Fc, a scFv, a scFv-Fc, a (scFv)2-Fc, an Fv, a single domain antibody (sdAb) (e.g., a VHH), a sdAb-Fc (e.g., a VHH-Fc), a (sdAb)2 (e.g., a (VHH)2), or a (sdAb)2-Fc (e.g., a (VHH)2-Fc).

3. The conjugate of any one of the preceding claims, wherein the antibody is a full-length antibody.

4. The conjugate of any one of the preceding claims, wherein the antibody is an IgG (e.g., a human IgG (hIgG)) antibody.

5. The conjugate of any one of the preceding claims, wherein the antibody is a hIgG1, hIgG2, hIgG3, or hIgG4 antibody (e.g., a hIgG1 or hIgG4 antibody).

6. The conjugate of any one of the preceding claims, wherein the antibody (e.g., full-length antibody) comprises an immunoglobulin (Ig) (e.g., a human Ig (hIg)) Fc region.

7. The conjugate of any one of the preceding claims, wherein the Ig (e.g., hIg) Fc region comprises at least a portion of a hinge region, a CH2 region, and a CH3 region.

8. The conjugate of any one of the preceding claims, wherein the Ig (e.g., hIg) Fc region comprises a hinge region, a CH2 region, and a CH3 region.

9. The conjugate of any one of the preceding claims, wherein the Ig is a hIg.

10. The conjugate of claim 9, wherein the hIg is a human IgG (hIgG).

11. The conjugate of claim 10, wherein the hIgG is hIgG1 or hIgG4.

12. The conjugate of any one of the preceding claims, wherein the Ig (e.g., hIg) Fc region comprises one or more amino acid substitutions relative to a reference Ig (e.g., hIg) Fc region that reduces or abolishes one or more of the following effector functions relative to the reference hIg Fc region: antibody dependent cell mediated cytotoxicity (ADCC), complement dependent cytotoxicity (CDC), and / or affinity to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))).Attorney Docket No.63340.10WO01 13. The conjugate of any one of the preceding claims, wherein the Ig (e.g., hIg) Fc region does not substantially mediate ADCC, does not substantially mediate CDC, and / or does not bind to one or more human Fc receptor (e.g., an Fcγ receptor (e.g., FcγRI, FcγRIIa, FcγRIIc, FcγRIIIa, and / or FcγRIIIb (e.g., FcγRI, FcγIIa, and / or FcγIIIa))).

14. The conjugate of any one of the preceding claims, wherein the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an amino acid substitution at amino acid position L234, and / or an amino acid substitution at amino acid position L235, numbering according to the EU index of Kabat.

15. The conjugate of any one of the preceding claims, wherein the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234 and / or an alanine at amino acid position L235, numbering according to the EU index of Kabat.

16. The conjugate of any one of the preceding claims, wherein the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234, an alanine at amino acid position L235, and / or a glycine, an alanine, or a serine at position P329 numbering according to the EU index of Kabat.

17. The conjugate of any one of the preceding claims, wherein the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position L234, a serine at amino acid position L235, and / or a glycine, an alanine, or a serine at position P329 numbering according to the EU index of Kabat.

18. The conjugate of any one of the preceding claims, wherein the Ig is hIgG1 and the amino acid sequence of the Fc region comprises an alanine at amino acid position N297, numbering according to the EU index of Kabat.

19. The conjugate of any one of the preceding claims, wherein the Ig is hIgG4 and the amino acid sequence of the Fc region comprises an amino acid substitution at amino acid position S228, an amino acid substitution at amino acid position F234, and / or an amino acid substitution at amino acid position L235, numbering according to the EU index of Kabat.

20. The conjugate of any one of the preceding claims, wherein the Ig is hIgG4 and the amino acid sequence of the Fc region comprises a proline at amino acid position S228, an alanine at amino acid position F234, and / or an alanine at amino acid position L235, numbering according to EU index of Kabat.Attorney Docket No.63340.10WO01 21. The conjugate any one of the preceding claims, wherein the Ig is hIgG4 and the amino acid sequence of the Fc region comprises an alanine at amino acid position N297, numbering according to the EU index of Kabat.

22. The conjugate of any one of the preceding claims, wherein the antibody comprises a first Fc region and a second Fc region that associate via at least one covalent (e.g., disulfide) bond.

23. The conjugate of any one of the preceding clams, wherein the antibody comprises a variable heavy chain (VH) region that comprises three complementarity determining regions: VH CDR1, VH CDR2, and VH CDR3; and a variable light chain (VL) region that comprises three complementarity determining regions: VL CDR1, VL CDR2, and VL CDR3.

24. The method of any one of the preceding claims, wherein (a) the amino acid sequence of VH CDR1 comprises the amino acid sequence SEQ ID NO: 3, or the amino acid sequence of SEQ ID NO: 3 comprising 1, 2, or 3 amino acid variations; (b) the amino acid sequence of VH CDR2 comprises the amino acid sequence SEQ ID NO: 4, or the amino acid sequence of SEQ ID NO: 4 comprising 1, 2, or 3 amino acid variations; (c) the amino acid sequence of VH CDR3 comprises the amino acid sequence SEQ ID NO: 5, or the amino acid sequence of SEQ ID NO: 5 comprising 1, 2, or 3 amino acid variations; and (d) the amino acid sequence of VL CDR1 comprises the amino acid sequence SEQ ID NO: 6, or the amino acid sequence of SEQ ID NO: 6 comprising 1, 2, or 3 amino acid variations; (e) the amino acid sequence of VL CDR2 comprises the amino acid sequence SEQ ID NO: 7, or the amino acid sequence of SEQ ID NO: 7 comprising 1, 2, or 3 amino acid variations; and (f) the amino acid sequence of VL CDR3 comprises the amino acid sequence SEQ ID NO: 8, or the amino acid sequence of SEQ ID NO: 8 comprising 1, 2, or 3 amino acid variations.

25. The method of any one of the preceding claims, wherein the VH region comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 9; and theAttorney Docket No.63340.10WO01 VL region comprises an amino acid sequence at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:

10.

26. The method of any one of the preceding claims, wherein the first moiety comprises a heavy chain (HC) and a light chain (LC), wherein (a) the amino acid sequence of the HC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 11; and the amino acid sequence of the LC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 12; or (b) the amino acid sequence of the HC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO: 13; and the amino acid sequence of the LC is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of SEQ ID NO:

12.

27. The conjugate of any one of the preceding claims, wherein the CpG ODN activates toll- like receptor 9 (TLR9) (i.e., the CpG oligonucleotide is a TLR9 agonist).

28. The conjugate of any one of the preceding claims, wherein the CpG ODN is a Class A, Class B, or Class C CpG ODN.

29. The conjugate of any one of the preceding claims, wherein the CpG ODN is a Class C CpG ODN.

30. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises a CpG ODN set forth in Table 8.

31. The conjugate of any one of the preceding claims, wherein the nucleotide sequence of the CpG ODN comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in Table 8; or comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in Table 8 comprising 1 or more (e.g., 1, 2, 3) but less than 15% (e.g., less than 12%, less than 10%, less than 8%) nucleotide variations (e.g., substitutions, additions, deletions, etc.).

32. The conjugate of any one of the preceding claims, wherein the nucleotide sequence of the CpG ODN comprises or consists of the nucleotide sequence of any one of the CpG ODNs setAttorney Docket No.63340.10WO01 forth in any one of SEQ ID NOS: 122-153 (e.g., SEQ ID NO: 122 or 123); or comprises or consists of the nucleotide sequence of any one of the CpG ODNs set forth in any one of SEQ ID NOS: 122-153 (e.g., SEQ ID NO: 122 or 123) and comprising 1 or more (e.g., 1, 2, 3) but less than 15% (e.g., less than 12%, less than 10%, less than 8%) nucleotide variations (e.g., substitutions, additions, deletions, etc.).

33. The conjugate of any one of the preceding claims, wherein the CpG ODN is single stranded.

34. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises from about 15-30, 15-25, 15-20, 16-30, 16-25, 16-20, 17-30, 17-25, 17-20, 18-30, 18-25, 18-20, 19-30, 19-25, or 19-20 nucleotides in length.

35. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises at least about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.

36. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises or consists of about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides.

37. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, or more CpG motifs.

38. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises one or more phosphodiester, phosphorothioate, phosphorodithioate, methylphosphonate, or methylphosphorothioate internucleoside linkage.

39. The conjugate of any one of the preceding claims, wherein the CpG ODN comprises one or more phosphorothioate, phosphorodithioate, methylphosphonate, or methylphosphorothioate internucleoside linkage.

40. The conjugate of any one of the preceding claims, wherein all of the internucleotide linkages of the CpG ODN are phosphorothioate linkages.

41. The conjugate of any one of the preceding claims, wherein the antibody is non-covalently conjugated to the at least one CpG ODN.

42. The conjugate of any one of the preceding claims, wherein the antibody is covalently conjugated to the at least one CpG ODN.

43. The conjugate of any one of the preceding claims, wherein the antibody is directly conjugated to the at least one CpG ODN.Attorney Docket No.63340.10WO01 44. The conjugate of any one of the preceding claims, wherein the antibody is indirectly conjugated to the at least one CpG ODN through a linker.

45. The conjugate of any one of the preceding claims, wherein the linker is cleavable or non- cleavable.

46. The conjugate of any one of the preceding claims, wherein the conjugate comprises at least 2, 3, 4, 5, 6, or more CpG ODNs.

47. The conjugate of any one of the preceding claims, wherein each of the at least 2, 3, 4, 5, 6, or more CpG ODNs are individually conjugated to the antibody.

48. The conjugate of any one of the preceding claims, wherein the conjugate activates a cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell))) upon binding CLEC2D expressed on the surface of the cell and / or internalization of the conjugate into the cell.

49. The conjugate of any one of the preceding claims, wherein upon internalization of the conjugate into an endosome of a cell expressing CLEC2D the conjugate activates toll-like receptor 9 (TLR9) expressed in the endosome.

50. The conjugate of any one of claims 3-33, wherein the conjugate inhibits (or substantially inhibits) binding of (e.g., hCD161) to CLEC2D (e.g., hCLEC2D).

51. A cell comprising the conjugate of any one of claims 1-50.

52. The cell of claim 50, wherein the cell is in vitro, ex vivo, or in vivo.

53. A pharmaceutical composition comprising the conjugate of any one of claims 1-50 and a pharmaceutically acceptable excipient.

54. A kit comprising the conjugate of any one of claims 1-50 or the pharmaceutical composition of any one of claims 53, and optionally instructions for utilizing any one of the foregoing.

55. A method of delivering a conjugate or pharmaceutical composition to a cell, the method comprising introducing into a cell the conjugate of any one of claims 1-50 or the pharmaceutical composition of claim 73, to thereby deliver the conjugate or pharmaceutical composition into the cell.

56. The method of claim 55, wherein the cell is in vitro, ex vivo, or in vivo.

57. The method of claim 55 or 56, wherein the cell is a subject (e.g., a human subject).Attorney Docket No.63340.10WO01 58. A method of delivering a conjugate, cell, or pharmaceutical composition to a subject, the method comprising administering to the subject the conjugate of any one of claims 1-50, the cell of any one of claims 51-52, or the pharmaceutical composition of claim 53, to thereby deliver the conjugate, cell, or pharmaceutical composition to the subject.

59. A method of activating an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-50, the cell of any one of claims 51-52, or the pharmaceutical composition of claim 53, to thereby activate an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject.

60. A method of activating TLR9 expressed by an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject, the method comprising administering to the subject the conjugate of any one of claims 1-50, the cell of any one of claims 51-52, or the pharmaceutical composition of claim 53, to thereby activate TLR9 expressed by an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in the subject.

61. A method of inhibiting binding of CD161 to CLEC2D expressed on the surface of an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in a subject, the method comprising administering to the subject the conjugate of any one of claims 1- 50, the cell of any one of claims 51-52, or the pharmaceutical composition of claim 53, to thereby inhibit binding of CD161 to CLEC2D expressed on the surface of an immune cell (e.g., a CLEC2D expressing immune cell (e.g., a B cell, a myeloid cell (e.g., macrophage, dendritic cell (e.g., plasmacytoid dendritic cell, classical dendritic cell)))) in the subject.

62. A method of treating a cancer (e.g., solid tumor) in a subject in need thereof, the method comprising administering to the subject the conjugate of any one of claims 1-50, the cell of any one of claims 51-52, or the pharmaceutical composition of claim 53, to thereby treat the cancer in the subject.

63. The method of claim 62, wherein the cancer is a solid tumor.Attorney Docket No.63340.10WO01 64. The method of any one of claims 62-63, wherein the cancer is a carcinoma.

65. The method of any one of claims 62-64, wherein the cancer is head cancer, neck cancer, head and neck cancer, lung cancer, colon cancer, rectal cancer, colorectal cancer, renal cancer, or ovarian cancer.

66. The method of any one of claims 62-65, wherein the cancer is head and neck squamous cell carcinoma (HNSCC), non-small cell lung cancer, colorectal carcinoma, or renal cell carcinoma.

67. The method of any one of claims 55-66, wherein the cell is a subject (e.g., a human subject).

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