Antibody molecules that bind to nkp30 and uses thereof

Abstract

Provides herein are compositions comprising an agent that comprises an antigen binding domain that binds to NKp30, and methods of treating conditions or diseases in a subject and methods of expanding and / or activation NK cells using the same.

Description

WSGR Docket No.53676-767.601 ANTIBODY MOLECULES THAT BIND TO NKP30 AND USES THEREOF CROSS REFERENCE

[0001] This application claims the benefit of U.S. provisional Application No.63 / 589,149, filed on October 10, 2023, which is incorporated herein by reference in its entirety. BACKGROUND

[0002] Natural Killer (NK) cells recognize and destroy tumors and virus-infected cells in an antibody- independent manner. The regulation of NK cells is mediated by activating and inhibiting receptors on the NK cell surface. One family of activating receptors is the natural cytotoxicity receptors (NCRs) which include NKp30, NKp44 and NKp46.

[0003] Given the importance of immune checkpoint pathways in regulating an immune response, the need exists for developing novel agents that modulate the activity of immunoinhibitory proteins, such as PD-1, thus leading to activation of immune system. Such agents can be used, e.g., for cancer immunotherapy and treatment of other conditions, such as chronic infection. SUMMARY

[0004] In an aspect, provided herein is, inter alia, a composition comprising an agent that comprises an antigen binding domain that binds to NKp30, wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising: (i) a heavy chain complementarity determining region 3 (VHCDR3) comprising the sequence of GDWHYFDY (SEQ ID NO: 504); and (ii) a heavy chain complementarity determining region 1 (VHCDR1) comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), a heavy chain complementarity determining region 2 (VHCDR2) comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), or a combination thereof.

[0005] In some embodiments, the antigen binding domain comprises a VH comprising the VHCDR1 comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), the VHCDR2 comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), and the VHCDR3 comprising the sequence of GDWHYFDY (SEQ ID NO: 504).

[0006] In some embodiments, the antigen binding domain comprises a light chain variable region (VL) comprising a light chain complementarity determining region 1 (VLCDR1) comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a light chain complementarity determining region 2 (VLCDR2) comprising the sequence of ENDRRPS (SEQ ID NO: 28), a light chain complementarity determining region 3 (VLCDR3) comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86), or any combination thereof.

[0007] In some embodiments, the antigen binding domain comprises a VL comprising a VLCDR1 comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a VLCDR2 comprising the sequence of ENDRRPS (SEQ ID NO: 28), and a VLCDR3 comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86).WSGR Docket No.53676-767.601

[0008] In some embodiments, the antigen binding domain comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 505.

[0009] In some embodiments, the antigen binding domain comprises a VH comprising the sequence of SEQ ID NO: 505.

[0010] In some embodiments, the antigen binding domain comprises a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 13.

[0011] In some embodiments, the antigen binding domain comprises a VL comprising the sequence of SEQ ID NO: 13.

[0012] In some embodiments, the antigen binding domain comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 505 and a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 13.

[0013] In some embodiments, the antigen binding domain comprises a VH comprising the sequence of SEQ ID NO: 505 and a VL comprising the sequence of SEQ ID NO: 13.

[0014] In some embodiments, the antigen binding domain is a full antibody or antigen-binding fragment thereof, a bivalent antibody, bispecific antibody, or a biparatopic antibody.

[0015] In some embodiments, the antigen binding domain is a Fab, F(ab')2, Fv, an single chain Fv (scFv), a diabody, a single domain antibody, or a camelid antibody.

[0016] In some embodiments, the antigen binding domain is a Fab or an scFv.

[0017] In some embodiments, the antigen binding domain comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or functional fragment thereof, IgG2 heavy chain constant region or functional fragment thereof, IgG3 heavy chain constant region or functional fragment thereof, IgGA1 heavy chain constant region or functional fragment thereof, IgGA2 heavy chain constant region or functional fragment thereof, IgG4 heavy chain constant region or functional fragment thereof, IgJ heavy chain constant region or functional fragment thereof, IgM heavy chain constant region or functional fragment thereof, IgD heavy chain constant region or functional fragment thereof, and IgE heavy chain constant region or functional fragment thereof.

[0018] In some embodiments, the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to any one heavy chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

[0019] In some embodiments, the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO:WSGR Docket No.53676-767.601 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, or the sequence of SEQ ID NO: 511.

[0020] In some embodiments, the antigen binding domain comprises a kappa light chain constant region or functional fragment thereof, a lambda light chain constant region or functional fragment thereof, or a combination thereof.

[0021] In some embodiments, the kappa light chain constant region comprises a sequence having at least 75% sequence identity to any one light chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

[0022] In some embodiments, the kappa light chain constant region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 39 or the sequence of SEQ ID NO: 3644.

[0023] In some embodiments, the agent is a polypeptide molecule.

[0024] In some embodiments, the agent is a multifunctional molecule.

[0025] In some embodiments, the agent is a multispecific molecule.

[0026] In some embodiments, the agent comprises at least one cytokine molecule or a functional fragment or variant thereof or a functional fragment or variant thereof, an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), a second antigen binding domain that binds to NKp30, an immune cell engager, a tumor-targeting moiety, or any combination thereof.

[0027] In some embodiments, the agent comprises at least two non-contiguous polypeptide chains comprising a first polypeptide chain and a second polypeptide chain; wherein the first polypeptide chain comprises, from N-terminus to C-terminus, the following configuration: A-[a first member of a dimerization module]-C, And the second polypeptide chain comprises, from N-terminus to C-terminus, the following configuration: B-[a second member of a dimerization module]-D; wherein the first member of the dimerization module comprises a first immunoglobulin constant domain; and the second member of the dimerization module comprises a second immunoglobulin constant domain; wherein at least one of A, B, C, and D is the antigen binding domain; and wherein the first polypeptide chain and the second polypeptide chain dimerize via interaction between the first member of the dimerization module and the second member of the dimerization module.

[0028] In some embodiments, (i) A is the antigen binding domain, and B, C, and D are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor-targeting moiety; (ii) B is the antigen binding domain, and A, C, and D are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds toWSGR Docket No.53676-767.601 T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor-targeting moiety; (iii) C is the antigen binding domain, and A, B, and D are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor-targeting moiety; (iv) D is the antigen binding domain, and A, B, and C are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor-targeting moiety.

[0029] In some embodiments, the agent comprises at least one cytokine molecule or a functional fragment or variant thereof.

[0030] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof, IL-7 or a functional fragment or variant thereof, IL-12 or a functional fragment or variant thereof, IL-15 or a functional fragment or variant thereof, IL-18 or a functional fragment or variant thereof, IL-21 or a functional fragment or variant thereof, interferon gamma or a functional fragment or variant thereof, or any combination thereof.

[0031] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

[0032] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

[0033] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof.

[0034] In some embodiments, the IL-2 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

[0035] In some embodiments, the IL-2 or a functional fragment or variant thereof comprises a sequence the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

[0036] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof IL-15 or a functional fragment or variant thereof.

[0037] In some embodiments, the IL-15 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2170, SEQ ID NO: 3670, or SEQ ID NO: 3673.WSGR Docket No.53676-767.601

[0038] In some embodiments, the IL-15 or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2170, the sequence of SEQ ID NO: 3670, or the sequence of SEQ ID NO: 3673.

[0039] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof further comprises an IL15Ralpha dimerizing domain covalently linked the IL-15 or a functional fragment or variant thereof.

[0040] In some embodiments, the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or a functional fragment or variant thereof.

[0041] In some embodiments, the IL15Ralpha dimerizing domain comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3472.

[0042] In some embodiments, the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

[0043] In some embodiments, the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

[0044] In some embodiments, the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

[0045] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3474, SEQ ID NO: 3671, or SEQ ID NO: 3674.

[0046] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3474, the sequence of SEQ ID NO: 3671, or the sequence of SEQ ID NO: 3674.

[0047] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-7 or a functional fragment or variant thereof.

[0048] In some embodiments, the IL-7 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

[0049] In some embodiments, the IL-7 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

[0050] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-12 or a functional fragment or variant or variant thereof.

[0051] In some embodiments, the IL-12 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

[0052] In some embodiments, the IL-12 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

[0053] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-18 of a functional fragment or variant thereof.WSGR Docket No.53676-767.601

[0054] In some embodiments, the IL-18 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2192.

[0055] In some embodiments, the IL-18 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2192.

[0056] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-21 or a functional fragment or variant thereof.

[0057] In some embodiments, the IL-21 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

[0058] In some embodiments, the IL-21 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

[0059] In some embodiments, the agent comprises an antigen binding domain that binds to TRBC.

[0060] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1 or TRBC2.

[0061] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

[0062] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

[0063] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 7, 8, 16, 39, and 40.WSGR Docket No.53676-767.601

[0064] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 7, 8, 16, 39, and 40.

[0065] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

[0066] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

[0067] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 7, 8, 16, 39, and 40.

[0068] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 7, 8, 16, 39, and 40.

[0069] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 9A, 9B, 11, 12, 13, 14, 15, 17, 39, and 40; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40;WSGR Docket No.53676-767.601 (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

[0070] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

[0071] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 11, 15, 17, 39, and 40.

[0072] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 11, 15, 17, 39, and 40.

[0073] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

[0074] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; orWSGR Docket No.53676-767.601 (iv) any combination thereof.

[0075] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 11, 15, 17, 39, and 40.

[0076] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 11, 15, 17, 39, and 40.

[0077] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the scFv sequences listed in Tables 12, 15, 17, 39, and 40.

[0078] In some embodiments, the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising any one sequence selected from the group consisting of the scFv sequences listed in Tables 12, 15, 17, 39, and 40.

[0079] In some embodiments, the agent comprises a second antigen binding domain that binds to NKp30.

[0080] In some embodiments, the antigen binding domain and the second antigen binding domain that binds to NKp30 are same.

[0081] In some embodiments, the antigen binding domain and the second antigen binding domain that binds to NKp30 are different.

[0082] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a heavy chain variable region (VH) comprising: (i) a heavy chain complementarity determining region 3 (VHCDR3) comprising the sequence of GDWHYFDY (SEQ ID NO: 504); and (ii) a heavy chain complementarity determining region 1 (VHCDR1) comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), a heavy chain complementarity determining region 2 (VHCDR2) comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), or a combination thereof.

[0083] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising the VHCDR1 comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), the VHCDR2 comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), and the VHCDR3 comprising the sequence of GDWHYFDY (SEQ ID NO: 504).

[0084] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a light chain variable region (VL) comprising a light chain complementarity determining region 1 (VLCDR1) comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a light chain complementarity determining region 2 (VLCDR2) comprising the sequence of ENDRRPS (SEQ ID NO: 28), a light chain complementarity determining region 3 (VLCDR3) comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86), or any combination thereof.WSGR Docket No.53676-767.601

[0085] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising a VLCDR1 comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a VLCDR2 comprising the sequence of ENDRRPS (SEQ ID NO: 28), and a VLCDR3 comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86).

[0086] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 505.

[0087] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising the sequence of SEQ ID NO: 505.

[0088] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 13.

[0089] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising the sequence of SEQ ID NO: 13.

[0090] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 505 and a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 13.

[0091] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising the sequence of SEQ ID NO: 505 and a VL comprising the sequence of SEQ ID NO: 13.

[0092] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a heavy chain variable region (VH) comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; or (iv) any combination thereof.

[0093] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; or (iv) any combination thereof.WSGR Docket No.53676-767.601

[0094] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 25, 26, 16, and 17.

[0095] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 25, 26, 16, and 17.

[0096] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 22, 24, 25, 26, 16, and 17; or (iv) any combination thereof.

[0097] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 22, 24, 25, 26, 16, and 17; or (iv) any combination thereof.

[0098] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 25, 26, 16, and 17.

[0099] In some embodiments, the second antigen binding domain that binds to NKp30 comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 25, 26, 16, and 17.

[0100] In some embodiments, the agent comprises an immune cell engager.

[0101] In some embodiments, the immune cell engager is selected from the group consisting of a T cell engager, an NK cell engager, a B cell engager, a dendritic cell engager, and the macrophage cell engager.

[0102] In some embodiments, the agent comprises a T cell engager.

[0103] In some embodiments, the T cell engager comprises an antigen binding domain that binds to a T cell receptor beta chain region (TCRβV).WSGR Docket No.53676-767.601

[0104] In some embodiments, the antigen binding domain that binds to a T cell receptor beta chain region (TCRβV) binds to a TCRβV of any one T cell receptor beta chain (TCRβ) subfamily member selected from the group consisting of TCRβ subfamily members listed in Tables 29 and 42A.

[0105] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a heavy chain variable region (VH) comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

[0106] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

[0107] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

[0108] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

[0109] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51;WSGR Docket No.53676-767.601 (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

[0110] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

[0111] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

[0112] In some embodiments, the antigen binding domain that binds to a TCRβV comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

[0113] In some embodiments, the agent comprises a NK cell engager.

[0114] In some embodiments, the NK cell engager does not bind to NKp30.

[0115] In some embodiments, the NK cell engager comprise a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequences listed in Table 27.

[0116] In some embodiments, the NK cell engager comprise any one sequence selected from the group consisting of the sequences listed in Table 27.

[0117] In some embodiments, the agent comprises a tumor-targeting moiety.

[0118] In some embodiments, the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor-targeting moiety, or any combination thereof is a full antibody or antigen-binding fragment thereof, a bivalent antibody, bispecific antibody, or a biparatopic antibody.

[0119] In some embodiments, the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor-targeting moiety, or any combination thereof is a Fab, F(ab')2, Fv, an single chain Fv (scFv), a diabody, a single domain antibody, or a camelid antibody.

[0120] In some embodiments, the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor-targeting moiety, or any combination thereof is a Fab or an scFv.WSGR Docket No.53676-767.601

[0121] In some embodiments, the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor-targeting moiety, or any combination thereof comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or functional fragment thereof, IgG2 heavy chain constant region or functional fragment thereof, IgG3 heavy chain constant region or functional fragment thereof, IgGA1 heavy chain constant region or functional fragment thereof, IgGA2 heavy chain constant region or functional fragment thereof, IgG4 heavy chain constant region or functional fragment thereof, IgJ heavy chain constant region or functional fragment thereof, IgM heavy chain constant region or functional fragment thereof, IgD heavy chain constant region or functional fragment thereof, and IgE heavy chain constant region or functional fragment thereof.

[0122] In some embodiments, the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to any one heavy chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

[0123] In some embodiments, the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, or the sequence of SEQ ID NO: 511.

[0124] In some embodiments, the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor-targeting moiety, or any combination thereof comprises a kappa light chain constant region or functional fragment thereof, a lambda light chain constant region or functional fragment thereof, or a combination thereof.

[0125] In some embodiments, the kappa light chain constant region comprises a sequence having at least 75% sequence identity to any one light chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

[0126] In some embodiments, the kappa light chain constant region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 39 or the sequence of SEQ ID NO: 3644.

[0127] In some embodiments, the first member of the dimerization module comprises a first Fc region and the second member of the dimerization module comprises a second Fc region.

[0128] In some embodiments, the first Fc region is selected from the group consisting of an IgG1 Fc region or a functional fragment thereof, an IgG2 Fc region or a functional fragment thereof, an IgG3 Fc region or a functional fragment thereof, an IgGA1 Fc region or a functional fragment thereof, an IgGA2 Fc region or a functional fragment thereof, an IgG4 Fc region or a functional fragment thereof, an IgJ FcWSGR Docket No.53676-767.601 region or a functional fragment thereof, an IgM Fc region or a functional fragment thereof, an IgD Fc region or a functional fragment thereof, and an IgE Fc region or a functional fragment thereof.

[0129] In some embodiments, the second Fc region is selected from the group consisting of an IgG1 Fc region or a functional fragment thereof, an IgG2 Fc region or a functional fragment thereof, an IgG3 Fc region or a functional fragment thereof, an IgGA1 Fc region or a functional fragment thereof, an IgGA2 Fc region or a functional fragment thereof, an IgG4 Fc region or a functional fragment thereof, an IgJ Fc region or a functional fragment thereof, an IgM Fc region or a functional fragment thereof, an IgD Fc region or a functional fragment thereof, and an IgE Fc region or a functional fragment thereof.

[0130] In some embodiments, the first Fc region, the second Fc region, or a combination thereof comprises an Fc interface with one or more of: a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, wherein the dimerization of the first Fc region and the second Fc region is enhanced as indicated by a greater ratio of heteromultimer:homomultimer forms relative to a dimerization of Fc regions with a non-engineered interface.

[0131] In some embodiments, (i) the first Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering (b) a Cys at position 349 in a heavy chain constant region according to EU Numbering), (c) a Ser at position 366 in a heavy chain constant region according to EU Numbering), (d) an Ala at position 368 in a heavy chain constant region according to EU Numbering); and (e) a Val at position 407 in a heavy chain constant region according to EU Numbering); and (ii) the second Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering), (b) a Cys at position 354 in a heavy chain constant region according to EU Numbering), and (c) a Trp at position 366 in a heavy chain constant region according to EU Numbering).

[0132] In some embodiments, (i) the first Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering), (b) a Cys at position 354 in a heavy chain constant region according to EU Numbering), and (c) a Trp at position 366 in a heavy chain constant region according to EU Numbering); and (ii) the second Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering (b) a Cys at position 349 in a heavy chain constant region according to EU Numbering), (c) a Ser at position 366 in a heavy chain constant region according to EU Numbering), (d) an Ala at position 368 in a heavy chain constant region according to EU Numbering); and (e) a Val at position 407 in a heavy chain constant region according to EU Numbering).

[0133] In some embodiments, the first Fc region comprises a mutation that decreases Fc receptor binding to the agent relative to an agent without the mutation.WSGR Docket No.53676-767.601

[0134] In some embodiments, the first Fc region comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation, or a combination thereof according to EU Numbering.

[0135] In some embodiments, the second Fc region comprises a mutation that decreases Fc receptor binding to the agent relative to an agent without the mutation.

[0136] In some embodiments, the second Fc region comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation, or a combination thereof according to EU Numbering.

[0137] In some embodiments, the first Fc region comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, and the sequence of SEQ ID NO: 511.

[0138] In some embodiments, the second Fc region comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, and the sequence of SEQ ID NO: 511.

[0139] In some embodiments, the first Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, and the sequence of SEQ ID NO: 511.

[0140] In some embodiments, the second Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO:WSGR Docket No.53676-767.601 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, and the sequence of SEQ ID NO: 511.

[0141] In some embodiments, the first Fc region comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3477, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 515, and the sequence of SEQ ID NO: 516.

[0142] In some embodiments, the second Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3477, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 515, or the sequence of SEQ ID NO: 516.

[0143] In some embodiments, the first Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 3648, SEQ ID NO: 508, SEQ ID NO: 506, SEQ ID NO: 507, SEQ ID NO: 3477, SEQ ID NO: 511, SEQ ID NO: 515, or SEQ ID NO: 516.

[0144] In some embodiments, the second Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 3648, SEQ ID NO: 508, SEQ ID NO: 506, SEQ ID NO: 507, SEQ ID NO: 3477, SEQ ID NO: 511, SEQ ID NO: 515, or SEQ ID NO: 516.

[0145] In some embodiments, (i) the first Fc region comprises the sequence of SEQ ID NO: 506 or the sequence of SEQ ID NO: 507, and the second Fc region comprises the sequence of SEQ ID NO: 3648 or the sequence of SEQ ID NO: 508; or (ii) the first Fc region comprises the sequence of SEQ ID NO: 3447 or the sequence of SEQ ID NO: 511, and the second Fc region comprises the sequence of SEQ ID NO: 515 or the sequence of SEQ ID NO: 516.

[0146] In some embodiments, (i) the first Fc region comprises the sequence of SEQ ID NO: 3648 or the sequence of SEQ ID NO: 508, and the second Fc region comprises the sequence of SEQ ID NO: 506 or the sequence of SEQ ID NO: 507; or (ii) the first Fc region comprises the sequence of SEQ ID NO: 515 or the sequence of SEQ ID NO: 516, and the second Fc region comprises the sequence of SEQ ID NO: 3447 or the sequence of SEQ ID NO: 511.

[0147] In some embodiments, the agent comprises a first polypeptide, a second polypeptide, and at least one cytokine molecule or a functional fragment or variant thereof, wherein the first polypeptide and the second polypeptide are non-contiguous;WSGR Docket No.53676-767.601 wherein: (i) the first polypeptide comprises a first member of the dimerization module linked to: (a) the antigen binding domain comprising a VH and a VL, or (b) a first portion of the antigen binding domain comprising the VH, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising a VL, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module, and wherein the at least one cytokine molecule or a functional fragment or variant thereof is covalently linked to the second polypeptide or the first polypeptide.

[0148] In some embodiments, the agent comprises a first polypeptide, a second polypeptide, and at least one cytokine molecule or a functional fragment or variant thereof, wherein the first polypeptide and the second polypeptide are non-contiguous; wherein: (i) the first polypeptide comprises a first member of a dimerization module linked to a first portion of the antigen binding domain comprising the VH, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising the VL, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module, and wherein the at least one cytokine molecule or a functional fragment or variant thereof is covalently linked to the second polypeptide.

[0149] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof is covalently linked to the N-terminus of the second polypeptide, the C-terminus of the second polypeptide, or a combination thereof.

[0150] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof is within a single contiguous polypeptide chain of the second polypeptide.

[0151] In some embodiments, the first polypeptide comprises: (i) the antigen binding domain comprising the VH and the VL, wherein the antigen binding domain further comprises a heavy chain constant domain 1 (CH1) linked to the VH; or (ii) the first portion of the first antigen binding domain comprising the VH, wherein the first portion of the antigen binding domain further comprises a CH1 linked to the VH.

[0152] In some embodiments, the CH1 is linked to the C-terminus of the VH.

[0153] In some embodiments, the agent comprises: (i) the first polypeptide comprising the antigen binding domain that comprises the VH and the VL, wherein the antigen binding domain further comprises a light chain constant domain (CL) linked to the VL; or (ii) the first polypeptide comprising the first portion of the antigen binding domain and the third polypeptide comprising the second portion of the antigen binding domain, wherein the second portion of the antigen binding domain further comprises a CL linked to the VL.WSGR Docket No.53676-767.601

[0154] In some embodiments, the CL is linked to the C-terminus of the VL.

[0155] In some embodiments, the first member of the dimerization module is linked to the C-terminus of the antigen binding domain comprising the VH and the VL, or the C-terminus of the first portion of the first antigen binding domain comprising the VH.

[0156] In some embodiments, the second polypeptide comprises: (i) an antigen binding domain that binds TRBC, an second antigen binding domain that binds to NKp30, an immune cell engager, or an tumor-targeting moiety comprising a VH and a VL, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a heavy chain constant domain 1 (CH1) linked to the VH; or (ii) the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising the VH, wherein the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a CH1 linked to the VH.

[0157] In some embodiments, the CH1 is linked to the C-terminus of the VH.

[0158] In some embodiments, the agent comprises: (i) the second polypeptide comprising an antigen binding domain that binds TRBC, an second antigen binding domain that binds to NKp30, an immune cell engager, or an tumor-targeting moiety that comprises a VH and a VL, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a light chain constant domain (CL) linked to the VL; or (ii) the second polypeptide comprising the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety, wherein the agent further comprises a fourth polypeptide comprising the second portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety, wherein the fourth polypeptide is non-contiguous with the first polypeptide, the second polypeptide, and third polypeptide, and wherein the second portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a CL linked to the VL.

[0159] In some embodiments, the CL is linked to the C-terminus of the VL.

[0160] In some embodiments, the second member of the dimerization module is linked to the C-terminus of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising the VH and the VL, or the C- terminus of the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising the VH.WSGR Docket No.53676-767.601

[0161] In some embodiments, the agent further comprises: (i) a linker between the first member of the dimerization module and the antigen binding domain comprising a VH and a VL, or the first portion of the antigen binding domain comprising a VH; (ii) a linker between the second member of the dimerization module and an antigen binding domain that binds TRBC, an second antigen binding domain that binds to NKp30, an immune cell engager, or an tumor-targeting moiety comprising a VH and a VL, or the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising a VH; (iii) a linker between the VH and the VL of the antigen binding domain; (iv) a linker between the CH1 and the VH of the antigen binding domain; (v) a linker between the CL and the VL of the antigen binding domain; (vi) a linker between the VH and the VL of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety; (vii) a linker between the CH1 and the VH of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety; (viii) a linker between the CL and the VL of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety; (ix) a linker between the at least one cytokine molecule or a functional fragment or variant thereof and the first polypeptide, a linker between the at least one cytokine molecule or a functional fragment or variant thereof and the second polypeptide, or any combination thereof; or (x) any combination thereof.

[0162] In some embodiments, the linker is selected from the group consisting of a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, and a non-helical linker.

[0163] In some embodiments, the linker is the peptide linker comprises the sequence of SEQ ID NO: 3473, SEQ ID NO: 3308, or SEQ ID NO: 8291.

[0164] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof is linked to the N-terminus of the first polypeptide, the C-terminus of the first polypeptide, the N- terminus of the second polypeptide, the C-terminus of the second polypeptide, the N-terminus of the third polypeptide, the C-terminus of the third polypeptide, or any combination thereof.

[0165] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof is within a single contiguous polypeptide chain of the first polypeptide, the second polypeptide, or the third cytokine polypeptide to which the at least one cytokine molecule or a functional fragment or variant thereof is linked.

[0166] In some embodiments, the agent comprises: (i) the first polypeptide comprising the first portion of the dimerization module linked to the C-terminus of the first portion of the antigen binding domain; (ii) the second polypeptide comprising the second portion of the dimerization module;WSGR Docket No.53676-767.601 (iii) the third polypeptide comprising the second portion of the antigen binding domain; and (iv) the at least one cytokine molecule or a functional fragment or variant thereof covalently linked to the N-terminus of the second polypeptide.

[0167] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3472, the sequence of SEQ ID NO: 2170, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

[0168] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 2170 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3644.

[0169] In some embodiments, the second polypeptide chain comprises, from the N-terminus to the C- terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 3473 operatively linked to the sequence of SEQ ID NO: 2170 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508.

[0170] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3474, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

[0171] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3474 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3644.WSGR Docket No.53676-767.601

[0172] In some embodiments, the second polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3474 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508.

[0173] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3420; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 501.

[0174] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4702; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4703; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 501.

[0175] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3420; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 501.

[0176] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 4702; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3420; and

[0177] (iii) a third polypeptide comprising the sequence of SEQ ID NO: 4703.In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3472, the sequence of SEQ ID NO: 3670, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

[0178] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 3670 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; andWSGR Docket No.53676-767.601 (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3644.

[0179] In some embodiments, the second polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 3473 operatively linked to the sequence of SEQ ID NO: 3670 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508.

[0180] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3671, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

[0181] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3671 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, comprising the sequence of SEQ ID NO: 13, and the sequence of SEQ ID NO: 3644.

[0182] In some embodiments, the second polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3671 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508.

[0183] In some embodiments, the wherein the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3669; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 501.

[0184] In some embodiments, the wherein the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4702; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4708; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 501.WSGR Docket No.53676-767.601

[0185] In some embodiments, the wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3669; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 501.

[0186] In some embodiments, the wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 4702; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 4708; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 501.

[0187] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of 2270, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

[0188] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of 2270 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

[0189] In some embodiments, the second polypeptide comprises from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 2270 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508.

[0190] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3340; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 501.

[0191] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4702; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4706; andWSGR Docket No.53676-767.601 (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 501.

[0192] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3340; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 501.

[0193] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 4702; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 4706; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 501.

[0194] In some embodiments, the agent comprises a first polypeptide and a second polypeptide, wherein the first polypeptide and the second polypeptide are non-contiguous, and wherein: (i) the first polypeptide comprises a first member of the dimerization module linked to: (a) the antigen binding domain comprising a VH of the antigen binding domain and a VL of the antigen binding domain, or (b) a first portion of the antigen binding domain comprising the VH of the antigen binding domain, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising a VL of the antigen binding domain, wherein the third polypeptide is non- contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module linked to: (a) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC) comprising a VH and a VL, or (b) a first portion of the antigen binding domain that binds to TRBC comprising the VH of the antigen binding domain that binds to TRBC, wherein the agent further comprises a fourth polypeptide comprising a second portion of the antigen binding domain that binds to TRBC comprising a VL of the antigen binding domain that binds to TRBC, wherein the fourth polypeptide is non-contiguous with the first polypeptide, the second polypeptide, and the third polypeptide.

[0195] In some embodiments, the agent comprises a first polypeptide and a second polypeptide, wherein the first polypeptide and the second polypeptide are non-contiguous, wherein: (i) the first polypeptide comprises a first member of the dimerization module linked to a first portion of the antigen binding domain comprising the VH of the antigen binding domain, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising a VL of the antigen binding domain, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module linked to an antigen binding domain that binds to TRBC.WSGR Docket No.53676-767.601

[0196] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3447 or SEQ ID NO: 511; (ii) a third polypeptide comprising the sequence of SEQ ID NO: 8308 and the sequence of SEQ ID NO: 3644; and (iii) a second polypeptide chain comprising the sequence of SEQ ID NO: 517 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514; or a second polypeptide chain comprising the sequence of SEQ ID NO: 512 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.

[0197] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3447 or SEQ ID NO: 511; (ii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8308 operatively linked to the sequence of SEQ ID NO: 3644; and (iii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 517 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514; or a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 512 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.

[0198] In some embodiments, the first polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 8291 operatively linked to the sequence of SEQ ID NO: 13.

[0199] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 509 or the sequence of SEQ ID NO: 4704; (ii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 7380; and (iii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 7382, the sequence of SEQ ID NO: 4677, the sequence of SEQ ID NO: 7379, or the sequence of 4675.

[0200] In some embodiments, the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 509 or the sequence of SEQ ID NO: 4704; (ii) a third polypeptide comprising the sequence of SEQ ID NO: 7380; and (iii) a second polypeptide chain comprising the sequence of SEQ ID NO: 7382, the sequence of SEQ ID NO: 4677, the sequence of SEQ ID NO: 7379, or the sequence of SEQ ID NO: 4675.

[0201] In some embodiments, the agent comprises a first polypeptide chain comprising the sequence of SEQ ID NO: 505, the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3447 or SEQ ID NO: 511.WSGR Docket No.53676-767.601

[0202] In some embodiments, the agent comprises a first polypeptide chain comprising, from the N- terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 13 operatively linked to sequence of SEQ ID NO: 3447 or SEQ ID NO: 511.

[0203] In some embodiments, the first polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 8291 operatively linked to the sequence of SEQ ID NO: 13.

[0204] In some embodiments, the agent comprises a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 509.

[0205] In some embodiments, the agent comprises a first polypeptide chain comprising the sequence of SEQ ID NO: 509.

[0206] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 7419 and the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 7420 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514 or a second polypeptide chain comprising the sequence of SEQ ID NO: 7423 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.

[0207] In some embodiments, the agent comprises: (i) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 7419 operatively linked to the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 7420 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514 or a second polypeptide chain comprising the sequence of SEQ ID NO: 7423 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.

[0208] In some embodiments, the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 7439; and (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4680, the sequence of SEQ ID NO: 7438, the sequence of SEQ ID NO: 4681, or the sequence of SEQ ID NO: 7440.

[0209] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 7439; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 4680, the sequence of SEQ ID NO: 7438, the sequence of SEQ ID NO: 4681, or the sequence of SEQ ID NO: 7440.

[0210] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8012 and the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8011 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.WSGR Docket No.53676-767.601

[0211] In some embodiments, the agent comprises: (i) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8012 operatively linked to the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8011 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.

[0212] In some embodiments, the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8002; and (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8001.

[0213] In some embodiments, the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8002; and (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4682.

[0214] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8002; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8001.

[0215] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8002; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 4682.

[0216] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8014 and the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8013 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, a second polypeptide chain comprising the sequence of SEQ ID NO: 8299 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, or a second polypeptide chain comprising the sequence of SEQ ID NO: 8303 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.

[0217] In some embodiments, the agent comprises: (i) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8014 operatively linked to the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8013 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, a second polypeptide chain comprising the sequence of SEQ ID NO: 8299 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, or a second polypeptide chain comprising the sequence of SEQ ID NO: 8303 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514.WSGR Docket No.53676-767.601

[0218] In some embodiments, the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8005; and (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8004, the sequence of SEQ ID NO: 4685, the sequence of SEQ ID NO: 8007, the sequence of SEQ ID NO: 4686, the sequence of SEQ ID NO: 4687, or the sequence of SEQ ID NO: 8009.

[0219] In some embodiments, the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8005; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8004, the sequence of SEQ ID NO: 4685, the sequence of SEQ ID NO: 8007, the sequence of SEQ ID NO: 4686, the sequence of SEQ ID NO: 4687, or the sequence of SEQ ID NO: 8009.

[0220] In some embodiments, the agent further comprises at least one cytokine molecule or a functional fragment or variant thereof, wherein the at least one cytokine molecule or a functional fragment or variant thereof is linked to the first member of the dimerization module, the second member of the dimerization module, the antigen binding domain, the antigen binding domain that binds to TRBC, or any combination thereof.

[0221] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof, IL-7 or a functional fragment or variant thereof, IL-12 or a functional fragment or variant thereof, IL-15 or a functional fragment or variant thereof, IL-18 or a functional fragment or variant thereof, IL-21 or a functional fragment or variant thereof, interferon gamma or a functional fragment or variant thereof, or any combination thereof.

[0222] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

[0223] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

[0224] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof.

[0225] In some embodiments, the IL-2 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

[0226] In some embodiments, the IL-2 or a functional fragment or variant thereof comprises a sequence the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

[0227] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof IL-15 or a functional fragment or variant thereof.WSGR Docket No.53676-767.601

[0228] In some embodiments, the IL-15 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2170, SEQ ID NO: 3670, or SEQ ID NO: 3673.

[0229] In some embodiments, the IL-15 or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2170, the sequence of SEQ ID NO: 3670, or the sequence of SEQ ID NO: 3673.

[0230] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof further comprises an IL15Ralpha dimerizing domain covalently linked the IL-15 or a functional fragment or variant thereof.

[0231] In some embodiments, the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or a functional fragment or variant thereof.

[0232] In some embodiments, the IL15Ralpha dimerizing domain comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3472.

[0233] In some embodiments, the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

[0234] In some embodiments, the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

[0235] In some embodiments, the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

[0236] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3474, SEQ ID NO: 3671, or SEQ ID NO: 3674.

[0237] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3474, the sequence of SEQ ID NO: 3671, or the sequence of SEQ ID NO: 3674.

[0238] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-7 or a functional fragment or variant thereof.

[0239] In some embodiments, the IL-7 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

[0240] In some embodiments, the IL-7 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

[0241] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-12 or a functional fragment or variant or variant thereof.

[0242] In some embodiments, the IL-12 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.WSGR Docket No.53676-767.601

[0243] In some embodiments, the IL-12 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

[0244] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-18 of a functional fragment or variant thereof.

[0245] In some embodiments, the IL-18 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2192.

[0246] In some embodiments, the IL-18 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2192.

[0247] In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-21 or a functional fragment or variant thereof.

[0248] In some embodiments, the IL-21 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

[0249] In some embodiments, the IL-21 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

[0250] In another aspect, provided herein is a recombinant polynucleotide molecule comprising a polynucleotide sequence encoding the agent of the composition as provided herein.

[0251] In some embodiments, the recombinant polynucleotide molecule is an isolated recombinant polynucleotide molecule.

[0252] In another aspect, provided herein is a vector comprising the recombinant polynucleotide molecule as provided herein.

[0253] In another aspect, provided herein is a cell comprising the recombinant polynucleotide molecule as provided herein, or the vector as provided herein.

[0254] In another aspect, provided herein is a method of making the agent of the composition of any one as provided herein, comprising culturing the cell as provided herein under conditions suitable for expression of the agent.

[0255] In another aspect, provided herein is a pharmaceutical composition comprising the composition as provided herein, the recombinant polynucleotide molecule as provided herein, the vector as provided herein, or the cell as provided herein, and a pharmaceutically acceptable carrier, excipient, or diluent.

[0256] In another aspect, provided herein is a method of treating a condition or disease in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the composition as provided herein, the recombinant polynucleotide molecule as provided herein, the vector as provided herein, the cell as provided herein, the pharmaceutical composition as provided herein, or any combination thereof, thereby treating the condition or disease in the subject.

[0257] In some embodiments, the condition or disease is cancer.

[0258] In some embodiments, the cancer is leukemia or lymphoma.

[0259] In some embodiments, the cancer is selected from the group consisting of acquired immune deficiency syndrome (AIDS)-associated lymphoma, Angioimmunoblastic T-cell lymphoma, Adult T-cellWSGR Docket No.53676-767.601 leukemia / lymphoma, Burkitt lymphoma, Central nervous system (CNS) lymphoma, Diffuse large B-cell lymphoma (DLBCL), Lymphoblastic lymphoma, Mantle cell lymphoma (MCL), Peripheral T-cell lymphoma (PTCL), Transformed follicular and transformed mucosa-associated lymphoid tissue (MALT) lymphomas, Cutaneous T-cell lymphoma (mycosis fungoides and Sézary syndrome), Follicular lymphoma, Lymphoplasmacytic lymphoma / Waldenström macroglobulinemia, Marginal zone B-cell lymphoma, Gastric mucosa-associated lymphoid tissue (MALT) lymphoma, Chronic lymphocytic leukemia / small-cell lymphocytic lymphoma (CLL / SLL), Extranodal T- / NK-cell lymphoma, and Anaplastic large-cell lymphoma.

[0260] In some embodiments, peripheral T-cell lymphoma (PTCL) is Hepatosplenic T-cell lymphoma (HSGDTCL), Subcutaneous paniculitis-like T-cell lymphoma, or Enteropathy-associated T-cell lymphoma.

[0261] In some embodiments, the cutaneous T-cell lymphoma is mycosis fungoides and Sézary syndrome.

[0262] In some embodiments, the extranodal T- / NK-cell lymphoma is a nasal type.

[0263] In some embodiments, the anaplastic large-cell lymphoma is primary cutaneous anaplastic large- cell lymphoma or systemic anaplastic large-cell lymphoma.

[0264] In some embodiments, the cancer is a solid tumor.

[0265] In some embodiments, the solid tumor is selected from the group consisting of pancreatic cancer, breast cancer, colon cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, esophageal cancer, endometrial cancer, gastric cancer, head and neck cancer, kidney cancer, and prostate cancer.

[0266] In some embodiments, the pancreatic cancer is pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma.

[0267] In some embodiments, the lung cancer is small cell lung cancer (SCLC) or non-small cell lung cancer (NSCLC).

[0268] In another aspect, provided herein is a method of expanding and / or activating NK cells in a NK cell population comprising contacting the NK cell population with an effective amount of the composition as provided herein, the recombinant polynucleotide molecule as provided herein, the vector as provided herein, the cell as provided herein, the pharmaceutical composition as provided herein, or any combination thereof, thereby, expanding and / or activating the NK cells in the NK cell population.

[0269] In some embodiments, the expanding and / or activating NK cells is in vivo or ex vivo expansion and / or activation. BRIEF DESCRIPTION OF THE DRAWINGS

[0270] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:WSGR Docket No.53676-767.601

[0271] FIGs.1A-1D are schematic representations of exemplary formats and configurations of the agents as provided herein. In some embodiments, the agent comprises the antigen binding domain that bind to NKp30 (“NKp30”), and the antigen binding domain that binds to TRBC1 (“TRBC1”). In some embodiments, the antigen binding domain that bind NKp30 comprises an scFv. In some embodiments, the antigen binding domain that bind NKp30 comprises an Fab. In some embodiments, the antigen binding domain that binds to TRBC1 comprises an scFv. In some embodiments, the antigen binding domain that binds to TRBC1 comprises an Fab. FIG.1A depicts an agent comprising an anti-TRBC1 Fab, an anti-NKp30 scFv, and an Fc dimer. The Fc dimer comprises two Fc chains. The C-terminus of the heavy chain of the anti-TRBC1 Fab is fused to the N-terminus of one Fc chain. The anti-NKp30 scFv is fused to the N-terminus of the other Fc chain. FIG.1B depicts an agent comprising an anti-TRBC1 Fab, an anti-NKp30 Fab, and an Fc dimer. The Fc dimer comprises two Fc chains. The C-terminus of the heavy chain of the anti-TRBC1 Fab is fused to the N-terminus of one Fc chain. The C-terminus of the heavy chain of the anti-NKp30 Fab is fused to the N-terminus of the other Fc chain. FIG.1C depicts an agent comprising an anti-TRBC1 scFv, an anti-NKp30 Fab and an Fc dimer. The Fc dimer comprises two Fc chains. The anti-TRBC1 scFv is fused to the N-terminus of the other Fc chain. The C-terminus of the heavy chain of the anti-NKp30 Fab is fused to the N-terminus of one Fc chain. FIG.1D depicts an agent comprising an anti-NKp30 scFv, an anti-TRBC1 scFv, and an Fc dimer. The Fc dimer comprises two Fc chains. The anti-TRBC1 scFv is fused to the N-terminus of one Fc chain. The anti-NKp30 scFv is fused to the N-terminus of the other Fc chain. In some embodiments, the antigen binding domain that binds to NKp30 is linked to the C-terminus of the first member of the dimerization module or the C-terminus of the second member of the dimerization module. In some embodiments, the agent comprises more than one antigen binding domains that bind to NKp30, and the more than one antigen binding domains that bind to NKp30 are independently linked to the N-terminus of the first member of the dimerization module, the N-terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof. In some embodiments, the antigen binding domain that binds to TRBC1 is linked to the C-terminus of the first member of the dimerization module or the C-terminus of the second member of the dimerization module. In some embodiments, the agent comprises more than one antigen binding domains that bind to TRBC1, and the more than one antigen binding domains that bind to TRBC1 are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof.

[0272] FIGs.2A-2D are schematic representations of exemplary formats and configurations of the agents as provided herein. In some embodiments, the agent comprises the antigen binding domain that bind to NKp30 (“NKp30”), and the antigen binding domain that binds to TRBC2 (“TRBC2”). In some embodiments, the antigen binding domain that bind NKp30 comprises an scFv. In some embodiments, the antigen binding domain that bind NKp30 comprises an Fab. In some embodiments, the antigenWSGR Docket No.53676-767.601 binding domain that binds to TRBC2 comprises an scFv. In some embodiments, the antigen binding domain that binds to TRBC2 comprises an Fab. FIG.2A depicts an agent comprising an anti-TRBC2 Fab, an anti-NKp30 scFv, and an Fc dimer. The Fc dimer comprises two Fc chains. The C-terminus of the heavy chain of the anti-TRBC2 Fab is fused to the N-terminus of one Fc chain. The anti-NKp30 scFv is fused to the N-terminus of the other Fc chain. FIG.2B depicts an agent comprising an anti-TRBC2 Fab, an anti-NKp30 Fab, and an Fc dimer. The Fc dimer comprises two Fc chains. The C-terminus of the heavy chain of the anti-TRBC2 Fab is fused to the N-terminus of one Fc chain. The C-terminus of the heavy chain of the anti-NKp30 Fab is fused to the N-terminus of the other Fc chain. FIG.2C depicts an agent comprising an anti-TRBC2 ScFv, an anti-NKp30 Fab and an Fc dimer. The Fc dimer comprises two Fc chains. The anti-TRBC2 scFv is fused to the N-terminus of the other Fc chain. The C-terminus of the heavy chain of the anti-NKp30 Fab is fused to the N-terminus of one Fc chain. FIG.2D depicts an agent comprising an anti-NKp30 scFv, an anti-TRBC2 scFv, and an Fc dimer. The Fc dimer comprises two Fc chains. The anti-TRBC2 scFv is fused to the N-terminus of one Fc chain. The anti-NKp30 scFv is fused to the N-terminus of the other Fc chain. In some embodiments, the antigen binding domain that binds to NKp30 is linked to the C-terminus of the first member of the dimerization module or the C- terminus of the second member of the dimerization module. In some embodiments, the agent comprises more than one antigen binding domains that bind to NKp30, and the more than one antigen binding domains that bind to NKp30 are independently linked to the N-terminus of the first member of the dimerization module, the N-terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof. In some embodiments, the antigen binding domain that binds to TRBC2 is linked to the C-terminus of the first member of the dimerization module or the C-terminus of the second member of the dimerization module. In some embodiments, the agent comprises more than one antigen binding domain that binds to TRBC2, and the more than one antigen binding domain that binds to TRBC2 are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof.

[0273] FIGs.3A-3B are schematic representations of exemplary formats and configurations of the agents as provided herein. In some embodiments, the agent comprises the antigen binding domain that bind NKp30 (“Nkp30”) and at least one cytokine molecule or a functional fragment or variant thereof (“cytokine”). In some embodiments, the antigen binding domain that bind NKp30 comprises an scFv. In some embodiments, the antigen binding domain that bind NKp30 comprises an Fab. FIG.3A depicts an agent comprising an anti-NKp30 scFv, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The anti-NKp30 scFv is fused to the N-terminus of one Fc chain. The cytokine molecule is fused to the N-terminus of the other Fc chain. FIG.3B depicts an agent comprising an anti-NKp30 Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The C-terminus of the heavy chain of the anti-NKp30 Fab is fused to the N-terminus of one Fc chain. The cytokineWSGR Docket No.53676-767.601 molecule is fused to the N-terminus of the other Fc chain. In some embodiments, the antigen binding domain that binds to NKp30 is linked to the C-terminus of the first member of the dimerization module or the C-terminus of the second member of the dimerization module. In some embodiments, the agent comprises more than one antigen binding domains that bind to NKp30, and the more than one antigen binding domains that bind to NKp30 are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C- terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof. In some embodiments, the at least one cytokine molecule or a functional fragment or variant thereof is linked to the C-terminus of the first member of the dimerization module or the C-terminus of the second member of the dimerization module. In some embodiments, the agent comprises more than one cytokine molecule or a functional fragment or variant thereof, and the more than one cytokine molecule or a functional fragment or variant thereof are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof.

[0274] In some embodiments, the agent comprises the antigen binding domain that binds to NKp30, the antigen binding domain that binds to TRBC1, and the at least one cytokine molecule or a functional fragment or variant thereof. In some embodiments, the antigen binding domain that binds to NKp30, the antigen binding domain that binds to TRBC1, and the at least one cytokine molecule or a functional fragment or variant thereof are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof. In some embodiments, the agent comprises the antigen binding domain that binds to NKp30, the antigen binding domain that binds to TRBC2, and the at least one cytokine molecule or a functional fragment or variant thereof. In some embodiments, the antigen binding domain that binds to NKp30, the antigen binding domain that binds to TRBC2, and the at least one cytokine molecule or a functional fragment or variant thereof are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C-terminus of the first member of the dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof. In some embodiments, the agent comprises the antigen binding domain that binds to NKp30, the antigen binding domain that binds to TRBC1 and / or the antigen binding domain that binds to TRBC2, and the at least one cytokine molecule or a functional fragment or variant thereof. In some embodiments, the antigen binding domain that binds to NKp30, the antigen binding domain that binds to TRBC1 and / or the antigen binding domain that binds to TRBC2, and the at least one cytokine molecule or a functional fragment or variant thereof are independently linked to the N- terminus of the first member of the dimerization module, the N- terminus of the second member of the dimerization module, the C-terminus of the first member of theWSGR Docket No.53676-767.601 dimerization module, the C-terminus of the second member of the dimerization module, or any combination thereof. DETAILED DESCRIPTION Definitions

[0275] In some embodiments, the multifunctional molecule includes an immune cell engager. “An immune cell engager” refers to one or more binding specificities that bind and / or activate an immune cell, e.g., a cell involved in an immune response. In embodiments, the immune cell is chosen from a T cell, an NK cell, a B cell, a dendritic cell, and / or the macrophage cell. The immune cell engager can be an antibody molecule, a receptor molecule (e.g., a full length receptor, receptor fragment, or fusion thereof (e.g., a receptor-Fc fusion)), or a ligand molecule (e.g., a full length ligand, ligand fragment, or fusion thereof (e.g., a ligand-Fc fusion)) that binds to the immune cell antigen (e.g., the T cell, the NK cell antigen, the B cell antigen, the dendritic cell antigen, and / or the macrophage cell antigen). In embodiments, the immune cell engager specifically binds to the target immune cell, e.g., binds preferentially to the target immune cell. For example, when the immune cell engager is an antibody molecule, it binds to an immune cell antigen (e.g., a T cell antigen, an NK cell antigen, a B cell antigen, a dendritic cell antigen, and / or a macrophage cell antigen) with a dissociation constant of less than about 10 nM.

[0276] In some embodiments, the multifunctional molecule includes a cytokine molecule. As used herein, a “cytokine molecule” refers to full length, a fragment or a variant of a cytokine; a cytokine further comprising a receptor domain, e.g., a cytokine receptor dimerizing domain; or an agonist of a cytokine receptor, e.g., an antibody molecule (e.g., an agonistic antibody) to a cytokine receptor, that elicits at least one activity of a naturally-occurring cytokine. In some embodiments the cytokine molecule is chosen from interleukin-2 (IL-2), interleukin-7 (IL-7), interleukin-12 (IL-12), interleukin-15 (IL-15), interleukin-18 (IL-18), interleukin-21 (IL-21), or interferon gamma, or a fragment or variant thereof, or a combination of any of the aforesaid cytokines. The cytokine molecule can be a monomer or a dimer. In embodiments, the cytokine molecule can further include a cytokine receptor dimerizing domain. In other embodiments, the cytokine molecule is an agonist of a cytokine receptor, e.g., an antibody molecule (e.g., an agonistic antibody) to a cytokine receptor chosen from an IL-15Ra or IL- 21R.

[0277] As used herein, the term “molecule” as used in, e.g., antibody molecule, cytokine molecule, receptor molecule, includes full-length, naturally-occurring molecules, as well as variants, e.g., functional variants (e.g., truncations, fragments, mutated (e.g., substantially similar sequences) or derivatized form thereof), so long as at least one function and / or activity of the unmodified (e.g., naturally-occurring) molecule remains.

[0278] In some embodiments, the multifunctional molecule includes a stromal modifying moiety. A “stromal modifying moiety,” as used herein refers to an agent, e.g., a protein (e.g., an enzyme), that is capable of altering, e.g., degrading a component of, the stroma. In embodiments, the component of the stroma is chosen from, e.g., an ECM component, e.g., a glycosaminoglycan, e.g., hyaluronan (alsoWSGR Docket No.53676-767.601 known as hyaluronic acid or HA), chondroitin sulfate, chondroitin, dermatan sulfate, heparin sulfate, heparin, entactin, tenascin, aggrecan and keratin sulfate; or an extracellular protein, e.g., collagen, laminin, elastin, fibrinogen, fibronectin, and vitronectin.

[0279] Certain terms are defined below.

[0280] As used herein, the articles “a” and “an” refer to one or more than one, e.g., to at least one, of the grammatical object of the article. The use of the words “a” or “an” when used in conjunction with the term “comprising” herein may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.”

[0281] As used herein, “about” and “approximately” generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 20 percent (%), typically, within 10%, and more typically, within 5% of a given range of values.

[0282] “Antibody molecule” as used herein refers to a protein, e.g., an immunoglobulin chain or fragment thereof, comprising at least one immunoglobulin variable domain sequence. An antibody molecule encompasses antibodies (e.g., full-length antibodies) and antibody fragments. In an embodiment, an antibody molecule comprises an antigen binding or functional fragment of a full length antibody, or a full length immunoglobulin chain. For example, a full-length antibody is an immunoglobulin (Ig) molecule (e.g., an IgG antibody) that is naturally occurring or formed by normal immunoglobulin gene fragment recombinatorial processes). In embodiments, an antibody molecule refers to an immunologically active, antigen-binding portion of an immunoglobulin molecule, such as an antibody fragment. An antibody fragment, e.g., functional fragment, is a portion of an antibody, e.g., Fab, Fab′, F(ab′)2, F(ab)2, variable fragment (Fv), domain antibody (dAb), or single chain variable fragment (scFv). A functional antibody fragment binds to the same antigen as that recognized by the intact (e.g., full-length) antibody. The terms “antibody fragment” or “functional fragment” also include isolated fragments consisting of the variable regions, such as the “Fv” fragments consisting of the variable regions of the heavy and light chains or recombinant single chain polypeptide molecules in which light and heavy variable regions are connected by a peptide linker (“scFv proteins”). In some embodiments, an antibody fragment does not include portions of antibodies without antigen binding activity, such as Fc fragments or single amino acid residues. Exemplary antibody molecules include full length antibodies and antibody fragments, e.g., dAb (domain antibody), single chain, Fab, Fab’, and F(ab’)2 fragments, and single chain variable fragments (scFvs).

[0283] As used herein, an “immunoglobulin variable domain sequence” refers to an amino acid sequence which can form the structure of an immunoglobulin variable domain. For example, the sequence may include all or part of the amino acid sequence of a naturally occurring variable domain. For example, the sequence may or may not include one, two, or more N- or C-terminal amino acids, or may include other alterations that are compatible with formation of the protein structure.

[0284] In embodiments, an antibody molecule is monospecific, e.g., it comprises binding specificity for a single epitope. In some embodiments, an antibody molecule is multispecific, e.g., it comprises a plurality of immunoglobulin variable domain sequences, where a first immunoglobulin variable domain sequenceWSGR Docket No.53676-767.601 has binding specificity for a first epitope and a second immunoglobulin variable domain sequence has binding specificity for a second epitope. In some embodiments, an antibody molecule is a bispecific antibody molecule. “Bispecific antibody molecule” as used herein refers to an antibody molecule that has specificity for more than one (e.g., two, three, four, or more) epitope and / or antigen.

[0285] “Antigen” (Ag) as used herein refers to a molecule that can provoke an immune response, e.g., involving activation of certain immune cells and / or antibody generation. Any macromolecule, including almost all proteins or peptides, can be an antigen. Antigens can also be derived from genomic recombinant or DNA. For example, any DNA comprising a nucleotide sequence or a partial nucleotide sequence that encodes a protein capable of eliciting an immune response encodes an “antigen.” In embodiments, an antigen does not need to be encoded solely by a full length nucleotide sequence of a gene, nor does an antigen need to be encoded by a gene at all. In embodiments, an antigen can be synthesized or can be derived from a biological sample, e.g., a tissue sample, a tumor sample, a cell, or a fluid with other biological components. As used, herein a “tumor antigen” or interchangeably, a “cancer antigen” includes any molecule present on, or associated with, a cancer, e.g., a cancer cell or a tumor microenvironment that can provoke an immune response. As used, herein an “immune cell antigen” includes any molecule present on, or associated with, an immune cell that can provoke an immune response.

[0286] The “antigen-binding site,” or “binding portion” of an antibody molecule refers to the part of an antibody molecule, e.g., an immunoglobulin (Ig) molecule, that participates in antigen binding. In embodiments, the antigen binding site is formed by amino acid residues of the variable (V) regions of the heavy (H) and light (L) chains. Three highly divergent stretches within the variable regions of the heavy and light chains, referred to as hypervariable regions, are disposed between more conserved flanking stretches called “framework regions,” (FRs). FRs are amino acid sequences that are naturally found between, and adjacent to, hypervariable regions in immunoglobulins. In embodiments, in an antibody molecule, the three hypervariable regions of a light chain and the three hypervariable regions of a heavy chain are disposed relative to each other in three dimensional space to form an antigen-binding surface, which is complementary to the three-dimensional surface of a bound antigen. The three hypervariable regions of each of the heavy and light chains are referred to as “complementarity-determining regions,” or “CDRs.” The framework region and CDRs have been defined and described, e.g., in Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.91-3242, and Chothia, C. et al. (1987) J. Mol. Biol.196:901-917, or by ImMunoGeneTics (IMGT) numbering system. Each variable chain (e.g., variable heavy chain and variable light chain) is typically made up of three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the amino acid order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4.

[0287] As used herein, the term “NKp30”, also known as Natural cytotoxicity triggering receptor 3, CD337 (cluster of differentiation 337), NCR3, 1C7, LY117, MALS, natural cytotoxicity triggering receptor 3, NCR3, nkp30, or NKP30, as used interchangeably herein, refers to the type I transmembrane receptor belonging to the immunoglobulin (Ig) superfamily that is expressed on resting and active naturalWSGR Docket No.53676-767.601 killer (NK) cells. In some embodiments, NKp30 can trigger cytotoxic response against tumor cells. The term “NKp30” includes isoforms, mammalian e.g., human NKp30, species homologs of human and analogs comprising at least one common epitope with NKp30. Human NKp30 comprises isoforms including, but not limited to : NKp30A, NKp30B, or NKp30C. Also, the term “NKp30” includes any of the recombinant or naturally-occurring forms of NKp30 or variants or homologs thereof that have or maintain NKp30 activity (e.g., at least 40% 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity). In some aspects, the variants or homologs have at least 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g., a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring NKp30. In some embodiments, NKp30 is substantially identical to the protein identified by the UniProt reference number O14931 or a variant or homolog having substantial identity thereto.

[0288] As used herein, the terms “T cell receptor beta variable chain,” “ T cell receptor beta chain variable region” “TCRβV,” “TCRβ V,” “TCR βV,” “TCRβv,” “TCR βv,” “TCRβ v,” “T cell receptor variable beta chain,” “TCRβV,” “TCR Vβ,” “TCRV β,” “TCRβV,” “TCRv β,” “TCR vβ,” “TCRBV,” “TCRVB,” “TRBV,” or “TCR beta V” are used interchangeably herein and refer to an extracellular region of the T cell receptor beta chain which comprises the antigen recognition domain of the T cell receptor. The term TCRβV includes isoforms, mammalian, e.g., human TCRβV, species homologs of human and analogs comprising at least one common epitope with TCRβV. Human TCRβV comprises a gene family comprising subfamilies including, but not limited to: a TCRβ V6 subfamily, a TCRβ V10 subfamily, a TCRβ V12 subfamily, a TCRβ V5 subfamily, a TCRβ V7 subfamily, a TCRβ V11 subfamily, a TCRβ V14 subfamily, a TCRβ V16 subfamily, a TCRβ V18 subfamily, a TCRβ V9 subfamily, a TCRβ V13 subfamily, a TCRβ V4 subfamily, a TCRβ V3 subfamily, a TCRβ V2 subfamily, a TCRβ V15 subfamily, a TCRβ V30 subfamily, a TCRβ V19 subfamily, a TCRβ V27 subfamily, a TCRβ V28 subfamily, a TCRβ V24 subfamily, a TCRβ V20 subfamily, TCRβ V25 subfamily, or a TCRβ V29 subfamily. In some embodiments, the TCRβ V6 subfamily comprises: TCRβ V6-4*01, TCRβ V6- 4*02, TCRβ V6-9*01, TCRβ V6-8*01, TCRβ V6-5*01, TCRβ V6-6*02, TCRβ V6-6*01, TCRβ V6- 2*01, TCRβ V6-3*01 or TCRβ V6-1*01. In some embodiments, TCRβV comprises TCRβ V6-5*01. TCRβ V6-5*01 is also known as TRBV65; TCRβV 6S5; TCRβV 13S1, or TCRβV 13.1. The amino acid sequence of TCRβ V6-5*01, e.g., human TCRβ V6-5*01, is known in that art, e.g., as provided by IMGT ID L36092.

[0289] “Cancer” as used herein can encompass all types of oncogenic processes and / or cancerous growths. In embodiments, cancer includes primary tumors as well as metastatic tissues or malignantly transformed cells, tissues, or organs. In embodiments, cancer encompasses all histopathologies and stages, e.g., stages of invasiveness / severity, of a cancer. In embodiments, cancer includes relapsed and / or resistant cancer. The terms “cancer” and “tumor” can be used interchangeably. For example, both terms encompass solid and liquid tumors. As used herein, the term “cancer” or “tumor” includes premalignant, as well as malignant cancers and tumors.WSGR Docket No.53676-767.601

[0290] As used herein, an “immune cell” refers to any of various cells that function in the immune system, e.g., to protect against agents of infection and foreign matter. In embodiments, this term includes leukocytes, e.g., neutrophils, eosinophils, basophils, lymphocytes, and monocytes. Innate leukocytes include phagocytes (e.g., macrophages, neutrophils, and dendritic cells), mast cells, eosinophils, basophils, and natural killer cells. Innate leukocytes identify and eliminate pathogens, either by attacking larger pathogens through contact or by engulfing and then killing microorganisms, and are mediators in the activation of an adaptive immune response. The cells of the adaptive immune system are special types of leukocytes, called lymphocytes. B cells and T cells are important types of lymphocytes and are derived from hematopoietic stem cells in the bone marrow. B cells are involved in the humoral immune response, whereas T cells are involved in cell-mediated immune response. The term “immune cell” includes immune effector cells.

[0291] “Immune effector cell,” as that term is used herein, refers to a cell that is involved in an immune response, e.g., in the promotion of an immune effector response. Examples of immune effector cells include, but are not limited to, T cells, e.g., alpha / beta T cells and gamma / delta T cells, B cells, natural killer (NK) cells, natural killer T (NK T) cells, and mast cells.

[0292] The term “effector function” or “effector response” refers to a specialized function of a cell. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines.

[0293] The compositions and methods of the present invention encompass polypeptides and nucleic acids having the sequences specified, or sequences substantially identical or similar thereto, e.g., sequences at least 80%, 85%, 90%, 95% identical or higher to the sequence specified. In the context of an amino acid sequence, the term “substantially identical” is used herein to refer to a first amino acid that contains a sufficient or minimum number of amino acid residues that are i) identical to, or ii) conservative substitutions of aligned amino acid residues in a second amino acid sequence such that the first and second amino acid sequences can have a common structural domain and / or common functional activity. For example, amino acid sequences that contain a common structural domain having at least about 80%, 85%, 90%.91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., a sequence provided herein.

[0294] In the context of nucleotide sequence, the term “substantially identical” is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity, or encode a common structural polypeptide domain or a common functional polypeptide activity. For example, nucleotide sequences having at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., a sequence provided herein.

[0295] The term “variant” refers to a polypeptide that has a substantially identical amino acid sequence to a reference amino acid sequence, or is encoded by a substantially identical nucleotide sequence. In some embodiments, the variant is a functional variant.WSGR Docket No.53676-767.601

[0296] The term “functional variant” refers to a polypeptide that has a substantially identical amino acid sequence to a reference amino acid sequence, or is encoded by a substantially identical nucleotide sequence, and is capable of having one or more activities of the reference amino acid sequence.

[0297] The term “scFv” refers to a fusion protein comprising at least one antibody fragment comprising a variable region of a light chain and at least one antibody fragment comprising a variable region of a heavy chain, wherein the light and heavy chain variable regions are contiguously linked via a short flexible polypeptide linker, and capable of being expressed as a single chain polypeptide, and wherein the scFv retains the specificity of the intact antibody from which it is derived. Unless specified, as used herein an scFv may have the VL and VH variable regions in either order, e.g., with respect to the N- terminal and C-terminal ends of the polypeptide, the scFv may comprise VL-linker-VH or may comprise VH-linker-VL.

[0298] The terms “complementarity determining region” or “CDR,” are used interchangeably herein and refer to the sequences of amino acids within antibody variable regions which confer antigen specificity and binding affinity. For example, in general, there are three CDRs in each heavy chain variable region (e.g., HCDR1, HCDR2, and HCDR3) and three CDRs in each light chain variable region (LCDR1, LCDR2, and LCDR3). The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of well-known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md. (“Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273,927-948 (“Chothia” numbering scheme), ImMunoGeneTics (IMGT) numbering system, or any combination thereof. Under the Kabat numbering scheme, in some embodiments, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3). Under the Chothia numbering scheme, in some embodiments, the CDR amino acids in the VHare numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95- 102 (HCDR3); and the CDR amino acid residues in the VLare numbered 26-32 (LCDR1), 50-52 (LCDR2), and 91-96 (LCDR3). In a combined Kabat and Chothia numbering scheme, in some embodiments, the CDRs correspond to the amino acid residues that are part of a Kabat CDR, a Chothia CDR, or both. For instance, in some embodiments, the CDRs correspond to amino acid residues 26-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3) in a VH, e.g., a mammalian VH, e.g., a human VH; and amino acid residues 24-34 (LCDR1), 50-56 (LCDR2), and 89-97 (LCDR3) in a VL, e.g., a mammalian VL, e.g., a human VL.

[0299] “Humanized” forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab’, F(ab’)2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin. For the most part, humanized antibodies and antibody fragments thereof are human immunoglobulins (recipient antibody or antibody fragment) in which residues from a complementary-determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody)WSGR Docket No.53676-767.601 such as mouse, rat or rabbit having the desired specificity, affinity, and capacity. In some instances, Fv framework region (FR) residues of the human immunoglobulin are replaced by corresponding non- human residues. Furthermore, a humanized antibody / antibody fragment can comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. These modifications can further refine and optimize antibody or antibody fragment performance. In general, the humanized antibody or antibody fragment thereof will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or a significant portion of the FR regions are those of a human immunoglobulin sequence. The humanized antibody or antibody fragment can also comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see Jones et al., Nature, 321: 522-525, 1986; Reichmann et al., Nature, 332: 323-329, 1988; Presta, Curr. Op. Struct. Biol., 2: 593-596, 1992.

[0300] “Fully human” refers to an immunoglobulin, such as an antibody or antibody fragment, where the whole molecule is of human origin or consists of an amino acid sequence identical to a human form of the antibody or immunoglobulin.

[0301] The term “specifically binds,” refers to an antibody, or a ligand, which recognizes and binds with a cognate binding partner (e.g., a stimulatory and / or costimulatory molecule present on a T cell) protein present in a sample, but which antibody or ligand does not substantially recognize or bind other molecules in the sample.

[0302] As used herein, an “immune cell” refers to any of various cells that function in the immune system, e.g., to protect against agents of infection and foreign matter. In embodiments, this term includes leukocytes, e.g., neutrophils, eosinophils, basophils, lymphocytes, and monocytes. Innate leukocytes include phagocytes (e.g., macrophages, neutrophils, and dendritic cells), mast cells, eosinophils, basophils, and natural killer cells. Innate leukocytes identify and eliminate pathogens, either by attacking larger pathogens through contact or by engulfing and then killing microorganisms, and are mediators in the activation of an adaptive immune response. The cells of the adaptive immune system are special types of leukocytes, called lymphocytes. B cells and T cells are important types of lymphocytes and are derived from hematopoietic stem cells in the bone marrow. B cells are involved in the humoral immune response, whereas T cells are involved in cell-mediated immune response. The term “immune cell” includes immune effector cells.

[0303] As used herein the term “immune effector cell,” refers to a cell that is involved in an immune response, e.g., in the promotion of an immune effector response. Examples of immune effector cells include, but are not limited to, T cells (e.g., alpha / beta T cells, gamma / delta T cells CD4+ T cells, CD8+ T cells), B cells, natural killer (NK) cells, natural killer T (NK T) cells, monocytes, macrophages, neutrophils, basophils, dendritic cells and mast cells.

[0304] The terms “effector function” or “effector response” refer to a specialized function of a cell. Effector function of a T cell, for example, may be cytolytic activity (e.g., CD8+ T cells) or helper activity (e.g., CD4+ T cells) including the secretion of cytokines.WSGR Docket No.53676-767.601

[0305] The term “antigen presenting cell” or “APC” refers to an immune system cell such as an accessory cell (e.g., a B-cell, a dendritic cell, and the like) that displays a foreign antigen complexed with major histocompatibility complexes (MHC’s) on its surface. T-cells may recognize these complexes using their T-cell receptors (TCRs). APCs process antigens and present them to T-cells.

[0306] The term, a “substantially purified cell” or “substantially purified cell population” refers to a cell or cell population that is essentially free of other cell types. A substantially purified cell also refers to a cell which has been separated from other cell types with which it is normally associated in its naturally occurring state. In some instances, a population of substantially purified cells refers to a homogenous population of cells. In other instances, this term refers simply to cell that have been separated from the cells with which they are naturally associated in their natural state. In some aspects, the cells are cultured in vitro. In other aspects, the cells are not cultured in vitro.

[0307] “Derived from” as that term is used herein, indicates a relationship between a first and a second molecule. It generally refers to structural similarity between the first molecule and a second molecule and does not connote or include a process or source limitation on a first molecule that is derived from a second molecule. For example, in the case of an intracellular signaling domain that is derived from a CD3zeta molecule, the intracellular signaling domain retains sufficient CD3zeta structure such that is has the required function, namely, the ability to generate a signal under the appropriate conditions. It does not connote or include a limitation to a particular process of producing the intracellular signaling domain, e.g., it does not mean that, to provide the intracellular signaling domain, one must start with a CD3zeta sequence and delete unwanted sequence, or impose mutations, to arrive at the intracellular signaling domain.

[0308] The term “encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (e.g., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene, cDNA, or RNA, encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

[0309] Calculations of homology or sequence identity between sequences (the terms are used interchangeably herein) are performed as follows.

[0310] To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length ofWSGR Docket No.53676-767.601 the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid “identity” is equivalent to amino acid or nucleic acid “homology”).

[0311] The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0312] The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In a preferred embodiment, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453 ) algorithm which has been incorporated into the GAP program in the GCG software package (available at gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In yet another preferred embodiment, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and the one that should be used unless otherwise specified) are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

[0313] The percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4:11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.

[0314] The nucleic acid and protein sequences described herein can be used as a “query sequence” to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol.215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to a nucleic acid molecule of the invention. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res.25:3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See ncbi.nlm.nih.gov.

[0315] It is understood that the molecules of the present invention may have additional conservative or non-essential amino acid substitutions, which do not have a substantial effect on their functions.

[0316] The term “amino acid” is intended to embrace all molecules, whether natural or synthetic, which include both an amino functionality and an acid functionality and capable of being included in a polymerWSGR Docket No.53676-767.601 of naturally occurring amino acids. Exemplary amino acids include naturally occurring amino acids; analogs, derivatives and congeners thereof; amino acid analogs having variant side chains; and all stereoisomers of any of any of the foregoing. As used herein the term “amino acid” includes both the D- or L- optical isomers and peptidomimetics.

[0317] A “conservative amino acid substitution” is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Such conservative modifications include amino acid substitutions, additions and deletions. Modifications can be introduced into an antibody or antibody fragment by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within a CAR can be replaced with other amino acid residues from the same side chain family and the altered CAR can be tested using the functional assays described herein

[0318] The terms “polypeptide”, “peptide” and “protein” (if single chain) are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids. The terms also encompass an amino acid polymer that has been modified; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation, such as conjugation with a labeling component. The polypeptide can be isolated from natural sources, can be a produced by recombinant techniques from a eukaryotic or prokaryotic host, or can be a product of synthetic procedures.

[0319] The terms “nucleic acid,” “nucleic acid sequence,” “nucleotide sequence,” or “polynucleotide sequence,” and “polynucleotide” are used interchangeably. They refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof. The polynucleotide may be either single-stranded or double-stranded, and if single-stranded may be the coding strand or non- coding (antisense) strand. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and nucleotide analogs. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component. The nucleic acid may be a recombinant polynucleotide, or a polynucleotide ofWSGR Docket No.53676-767.601 genomic, cDNA, semisynthetic, or synthetic origin which either does not occur in nature or is linked to another polynucleotide in a non-natural arrangement.

[0320] The term “isolated,” as used herein, refers to material that is removed from its original or native environment (e.g., the natural environment if it is naturally occurring). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or polypeptide, separated by human intervention from some or all of the co-existing materials in the natural system, is isolated. Such polynucleotides could be part of a vector and / or such polynucleotides or polypeptides could be part of a composition, and still be isolated in that such vector or composition is not part of the environment in which it is found in nature.

[0321] The term “endogenous” refers to any material from or produced inside an organism, cell, tissue or system.

[0322] The term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue or system.

[0323] The term “expression” refers to the transcription and / or translation of a particular nucleotide sequence driven by a promoter.

[0324] The term “transfer vector” refers to a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “transfer vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to further include non- plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, a polylysine compound, liposome, and the like. Examples of viral transfer vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

[0325] The term “expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, including cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.

[0326] The term “vector” as used herein refers to any vehicle that can be used to deliver and / or express a nucleic acid molecule. It can be a transfer vector or an expression vector as described herein.

[0327] The term “lentivirus” refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient methods of a gene delivery vector.WSGR Docket No.53676-767.601

[0328] The term “lentiviral vector” refers to a vector derived from at least a portion of a lentivirus genome, including especially a self-inactivating lentiviral vector as provided in Milone et al., Mol. Ther. 17(8): 1453-1464 (2009). Other examples of lentivirus vectors that may be used in the clinic, include but are not limited to, e.g., the LENTIVECTOR® gene delivery technology from Oxford BioMedica, the LENTIMAX™ vector system from Lentigen and the like. Nonclinical types of lentiviral vectors are also available and would be known to one skilled in the art.

[0329] The term “operably linked” or “transcriptional control” refers to functional linkage between a regulatory sequence and a heterologous nucleic acid sequence resulting in expression of the latter. For example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Operably linked DNA sequences can be contiguous with each other and, e.g., where necessary to join two protein coding regions, are in the same reading frame.

[0330] The term “parenteral” administration of an immunogenic composition includes, e.g., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrasternal injection, intratumoral, or infusion techniques.

[0331] The term “promoter” refers to a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the specific transcription of a polynucleotide sequence.

[0332] The term “promoter / regulatory sequence” refers to a nucleic acid sequence which is required for expression of a gene product operably linked to the promoter / regulatory sequence. In some instances, this sequence may be the core promoter sequence and in other instances, this sequence may also include an enhancer sequence and other regulatory elements which are required for expression of the gene product. The promoter / regulatory sequence may, for example, be one which expresses the gene product in a tissue specific manner.

[0333] The term “constitutive promoter” refers to a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell.

[0334] The term “inducible promoter” refers to a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell substantially only when an inducer which corresponds to the promoter is present in the cell.

[0335] The term “tissue-specific promoter” refers to a nucleotide sequence which, when operably linked with a polynucleotide encodes or specified by a gene, causes the gene product to be produced in a cell substantially only if the cell is a cell of the tissue type corresponding to the promoter.

[0336] As used herein, “transient” refers to expression of a non-integrated transgene for a period of hours, days or weeks, wherein the period of time of expression is less than the period of time for expression of the gene if integrated into the genome or contained within a stable plasmid replicon in the host cell.WSGR Docket No.53676-767.601

[0337] The term “transfected” or “transformed” or “transduced” refers to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A “transfected” or “transformed” or “transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny.

[0338] The term “chimeric antigen receptor” or alternatively a “CAR” are used interchangeably herein and refer to a recombinant polypeptide construct comprising at least an extracellular antigen binding domain, a transmembrane domain and a cytoplasmic signaling domain (also referred to herein as “an intracellular signaling domain”) comprising a functional signaling domain derived from a stimulatory molecule as defined below. In some embodiments, the domains in the CAR polypeptide construct are in the same polypeptide chain, e.g., comprise a chimeric fusion protein. In some embodiments, the domains in the CAR polypeptide construct are not contiguous with each other, e.g., are in different polypeptide chains. In one aspect, the stimulatory molecule of the CAR is the zeta chain associated with the T cell receptor complex. In one aspect, the cytoplasmic signaling domain comprises a primary signaling domain (e.g., a primary signaling domain of CD3-zeta). In one aspect, the cytoplasmic signaling domain further comprises one or more functional signaling domains derived from at least one costimulatory molecule as defined below. In one aspect, the costimulatory molecule is chosen from 4-1BB (i.e., CD137), CD27, ICOS, and / or CD28. In one aspect, the CAR comprises a chimeric fusion protein comprising an extracellular antigen recognition domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain derived from a stimulatory molecule. In one aspect, the CAR comprises a chimeric fusion protein comprising an extracellular antigen recognition domain, a transmembrane domain and an intracellular signaling domain comprising a functional signaling domain derived from a co-stimulatory molecule and a functional signaling domain derived from a stimulatory molecule. In one aspect, the CAR comprises a chimeric fusion protein comprising an extracellular antigen recognition domain, a transmembrane domain and an intracellular signaling domain comprising two functional signaling domains derived from one or more co-stimulatory molecule(s) and a functional signaling domain derived from a stimulatory molecule. In one aspect, the CAR comprises a chimeric fusion protein comprising an extracellular antigen recognition domain, a transmembrane domain and an intracellular signaling domain comprising at least two functional signaling domains derived from one or more co-stimulatory molecule(s) and a functional signaling domain derived from a stimulatory molecule. In one aspect the CAR comprises an optional leader sequence at the amino-terminus (N-ter) of the CAR fusion protein. In one aspect, the CAR further comprises a leader sequence at the N-terminus of the extracellular antigen recognition domain, wherein the leader sequence is optionally cleaved from the antigen recognition domain (e.g., a scFv) during cellular processing and localization of the CAR to the cellular membrane.

[0339] The term “signaling domain” as used herein refers to the functional portion of a protein which acts by transmitting information within the cell to regulate cellular activity via defined signaling pathways by generating second messengers or functioning as effectors by responding to such messengers.WSGR Docket No.53676-767.601

[0340] An “intracellular signaling domain,” as the term is used herein, refers to an intracellular portion of a molecule. The intracellular signaling domain can generate a signal that promotes an immune effector function of the CAR containing cell, e.g., a CART cell or CAR-expressing NK cell. Examples of immune effector function, e.g., in a CART cell or CAR-expressing NK cell, include cytolytic activity and helper activity, including the secretion of cytokines. In embodiments, the intracellular signal domain transduces the effector function signal and directs the cell to perform a specialized function. While the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal. The term intracellular signaling domain is thus meant to include any truncated portion of the intracellular signaling domain sufficient to transduce the effector function signal. In some embodiment, the intracellular signaling domain comprises a primary intracellular signaling domain. Exemplary primary intracellular signaling domains include those derived from the molecules responsible for primary stimulation, or antigen dependent simulation. In an embodiment, the intracellular signaling domain can comprise a costimulatory intracellular domain. Exemplary costimulatory intracellular signaling domains include those derived from molecules responsible for costimulatory signals, or antigen independent stimulation. For example, in the case of a CAR-expressing immune effector cell, e.g., CART cell or CAR-expressing NK cell, a primary intracellular signaling domain can comprise a cytoplasmic sequence of a T cell receptor, and a costimulatory intracellular signaling domain can comprise cytoplasmic sequence from co- receptor or costimulatory molecule. A primary intracellular signaling domain can comprise a signaling motif which is known as an immunoreceptor tyrosine-based activation motif or ITAM. Examples of ITAM containing primary cytoplasmic signaling sequences include, but are not limited to, those derived from CD3 zeta, FcR gamma, FcR beta, CD3 gamma, CD3 delta, CD3 epsilon, CDS, CD22, CD79a, CD79b, CD278 (“ICOS”), FcεRI, CD66d, DAP10, and DAP12.

[0341] The term “zeta” or alternatively “zeta chain”, “CD3-zeta” or “TCR-zeta” is defined as the protein provided as GenBan Acc. No. BAG36664.1, or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like, and a “zeta stimulatory domain” or alternatively a “CD3-zeta stimulatory domain” or a “TCR-zeta stimulatory domain” is defined as the amino acid residues from the cytoplasmic domain of the zeta chain that are sufficient to functionally transmit an initial signal necessary for T cell activation. In one aspect the cytoplasmic domain of zeta comprises residues 52 through 164 of GenBank Acc. No. BAG36664.1 or the equivalent residues from a non-human species, e.g., mouse, rodent, monkey, ape and the like, that are functional orthologs thereof.

[0342] The term “costimulatory molecule” refers to the cognate binding partner on a T cell that specifically binds with a costimulatory ligand, thereby mediating a costimulatory response by the T cell, such as, but not limited to, proliferation. Costimulatory molecules are cell surface molecules other than antigen receptors or their ligands that are required for an efficient immune response. Costimulatory molecules include, but are not limited to an a MHC class I molecule, TNF receptor proteins, Immunoglobulin-like proteins, cytokine receptors, integrins, signaling lymphocytic activation moleculesWSGR Docket No.53676-767.601 (SLAM proteins), activating NK cell receptors, BTLA, a Toll ligand receptor, OX40, CD2, CD7, CD27, CD28, CD30, CD40, CDS, ICAM-1, LFA-1 (CD11a / CD18), 4-1BB (CD137), B7-H3, CDS, ICAM-1, ICOS (CD278), GITR, BAFFR, LIGHT, HVEM (LIGHTR), KIRDS2, SLAMF7, NKp80 (KLRF1), NKp44, NKp30, NKp46, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD11a, LFA-1, ITGAM, CD11b, ITGAX, CD11c, ITGB1, CD29, ITGB2, CD18, LFA-1, ITGB7, NKG2D, NKG2C, TNFR2, TRANCE / RANKL, DNAM1 (CD226), SLAMF4 (CD244, 2B4), CD84, CD96 (Tactile), CEACAM1, CRTAM, Ly9 (CD229), CD160 (BY55), PSGL1, CD100 (SEMA4D), CD69, SLAMF6 (NTB-A, Ly108), SLAM (SLAMF1, CD150, IPO-3), BLAME (SLAMF8), SELPLG (CD162), LTBR, LAT, GADS, SLP-76, PAG / Cbp, CD19a, and a ligand that specifically binds with CD83.

[0343] A “costimulatory intracellular signaling domain” refers to the intracellular portion of a costimulatory molecule. The intracellular signaling domain can comprise the entire intracellular portion, or the entire native intracellular signaling domain, of the molecule from which it is derived, or a functional fragment thereof.

[0344] The term “signal transduction pathway” as used herein refers to the biochemical relationship between a variety of signal transduction molecules that play a role in the transmission of a signal from one portion of a cell to another portion of a cell.

[0345] The term “cell surface receptor” as used herein includes molecules and complexes of molecules capable of receiving a signal and transmitting signal across the membrane of a cell.

[0346] The term “anti-tumor effect” or “anti-cancer effect,” used interchangeably herein refer to a biological effect which can be manifested by various means, including but not limited to, e.g., a decrease in tumor volume, a decrease in the number of tumor cells, a decrease in the number of metastases, an increase in life expectancy, decrease in tumor cell proliferation, decrease in tumor cell survival, or amelioration of various physiological symptoms associated with the cancerous condition. An “anti-tumor effect” can also be manifested by the ability of the peptides, polynucleotides, cells and antibodies in prevention of the occurrence of tumor in the first place.

[0347] The terms “Cancer” or “tumor” as used interchangeably herein and encompass all types of oncogenic processes and / or cancerous growths. In embodiments, cancer includes primary tumors as well as metastatic tissues or malignantly transformed cells, tissues, or organs. In embodiments, cancer encompasses all histopathologies and stages, e.g., stages of invasiveness / severity, of a cancer. In embodiments, cancer includes relapsed and / or resistant cancer. For example, both terms encompass solid and liquid tumors. As used herein, the term cancer includes premalignant, as well as malignant cancers and tumors.

[0348] The term “autologous” refers to any material derived from the same individual to whom it is later to be re-introduced into the individual.

[0349] The term “allogeneic” refers to any material derived from a different animal of the same species as the individual to whom the material is introduced. Two or more individuals are said to be allogeneic toWSGR Docket No.53676-767.601 one another when the genes at one or more loci are not identical. In some aspects, allogeneic material from individuals of the same species may be sufficiently unlike genetically to interact antigenically.

[0350] The term “xenogeneic” refers to a graft derived from an animal of a different species.

[0351] The term “apheresis” as used herein refers to the art-recognized extracorporeal process by which the blood of a donor or patient is removed from the donor or patient and passed through an apparatus that separates out selected particular constituent(s) and returns the remainder to the circulation of the donor or patient, e.g., by re-transfusion. Thus, in the context of “an apheresis sample” refers to a sample obtained using apheresis.

[0352] The term “combination” refers to either a fixed combination in one dosage unit form, or a combined administration where a compound and a combination partner (e.g. another drug as explained below, also referred to as “therapeutic agent” or “co-agent”) may be administered independently at the same time or separately within time intervals, especially where these time intervals allow that the combination partners show a cooperative, e.g. synergistic effect. The single components may be packaged in a kit or separately. One or both of the components (e.g., powders or liquids) may be reconstituted or diluted to a desired dose prior to administration. The terms “co-administration” or “combined administration” or the like as utilized herein are meant to encompass administration of the selected combination partner to a single subject in need thereof (e.g. a patient), and are intended to include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time. The term “pharmaceutical combination” as used herein means a product that results from the mixing or combining of more than one active ingredient and includes both fixed and non-fixed combinations of the active ingredients. The term “fixed combination” means that the active ingredients, e.g. a compound and a combination partner, are both administered to a patient simultaneously in the form of a single entity or dosage. The term “non-fixed combination” means that the active ingredients, e.g. a compound and a combination partner, are both administered to a patient as separate entities either simultaneously, concurrently or sequentially with no specific time limits, wherein such administration provides therapeutically effective levels of the two compounds in the body of the patient. The latter also applies to cocktail therapy, e.g. the administration of three or more active ingredients.

[0353] The term “effective amount” or “therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result.

[0354] As used herein, the terms “treat,” “treatment,” and “treating” refer to the reduction or amelioration of the progression, severity and / or duration of a proliferative disorder, or the amelioration of one or more symptoms (preferably, one or more discernible symptoms) of a proliferative disorder resulting from the administration of one or more therapies (e.g., one or more therapeutic agents such as a CAR). In specific embodiments, the terms “treat,” “treatment,” and “treating” refer to the amelioration of at least one measurable physical parameter of a proliferative disorder, such as growth of a tumor, not necessarily discernible by the patient. In other embodiments the terms “treat”, “treatment” and “treating”-WSGR Docket No.53676-767.601 refer to the inhibition of the progression of a proliferative disorder, either physically by, e.g., stabilization of a discernible symptom, physiologically by, e.g., stabilization of a physical parameter, or both. In other embodiments the terms “treat,” “treatment,” and “treating” refer to the reduction or stabilization of tumor size or cancerous cell count.

[0355] The term “therapeutic” as used herein means a treatment. A therapeutic effect is obtained by reduction, suppression, remission, or eradication of a disease state.

[0356] The term “prophylaxis” as used herein means the prevention of or protective treatment for a disease or disease state.

[0357] The term “subject” is intended to include living organisms in which an immune response can be elicited (e.g., mammals, human).

[0358] Various aspects of the invention are described in further detail below. Additional definitions are set out throughout the specification. Antibody Molecules

[0359] In one embodiment, the antibody molecule binds to a cancer antigen, e.g., a tumor antigen or a stromal antigen. In some embodiments, the cancer antigen is, e.g., a mammalian, e.g., a human, cancer antigen. In other embodiments, the antibody molecule binds to an immune cell antigen, e.g., a mammalian, e.g., a human, immune cell antigen. For example, the antibody molecule binds specifically to an epitope, e.g., linear or conformational epitope, on the cancer antigen or the immune cell antigen.

[0360] In an embodiment, an antibody molecule is a monospecific antibody molecule and binds a single epitope. E.g., a monospecific antibody molecule having a plurality of immunoglobulin variable domain sequences, each of which binds the same epitope.

[0361] In an embodiment an antibody molecule is a multispecific or multifunctional antibody molecule, e.g., it comprises a plurality of immunoglobulin variable domains sequences, wherein a first immunoglobulin variable domain sequence of the plurality has binding specificity for a first epitope and a second immunoglobulin variable domain sequence of the plurality has binding specificity for a second epitope. In an embodiment the first and second epitopes are on the same antigen, e.g., the same protein (or subunit of a multimeric protein). In an embodiment the first and second epitopes overlap. In an embodiment the first and second epitopes do not overlap. In an embodiment the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein). In an embodiment a multispecific antibody molecule comprises a third, fourth or fifth immunoglobulin variable domain. In an embodiment, a multispecific antibody molecule is a bispecific antibody molecule, a trispecific antibody molecule, or a tetraspecific antibody molecule.

[0362] In an embodiment a multispecific antibody molecule is a bispecific antibody molecule. A bispecific antibody has specificity for no more than two antigens. A bispecific antibody molecule is characterized by a first immunoglobulin variable domain sequence which has binding specificity for a first epitope and a second immunoglobulin variable domain sequence that has binding specificity for a second epitope. In an embodiment the first and second epitopes are on the same antigen, e.g., the sameWSGR Docket No.53676-767.601 protein (or subunit of a multimeric protein). In an embodiment the first and second epitopes overlap. In an embodiment the first and second epitopes do not overlap. In an embodiment the first and second epitopes are on different antigens, e.g., the different proteins (or different subunits of a multimeric protein). In an embodiment a bispecific antibody molecule comprises a heavy chain variable domain sequence and a light chain variable domain sequence which have binding specificity for a first epitope and a heavy chain variable domain sequence and a light chain variable domain sequence which have binding specificity for a second epitope. In an embodiment a bispecific antibody molecule comprises a half antibody having binding specificity for a first epitope and a half antibody having binding specificity for a second epitope. In an embodiment a bispecific antibody molecule comprises a half antibody, or fragment thereof, having binding specificity for a first epitope and a half antibody, or fragment thereof, having binding specificity for a second epitope. In an embodiment a bispecific antibody molecule comprises a scFv or a Fab, or fragment thereof, have binding specificity for a first epitope and a scFv or a Fab, or fragment thereof, have binding specificity for a second epitope.

[0363] In an embodiment, an antibody molecule comprises a diabody, and a single-chain molecule, as well as an antigen-binding fragment of an antibody (e.g., Fab, F(ab’)2, and Fv). For example, an antibody molecule can include a heavy (H) chain variable domain sequence (abbreviated herein as VH), and a light (L) chain variable domain sequence (abbreviated herein as VL). In an embodiment an antibody molecule comprises or consists of a heavy chain and a light chain (referred to herein as a half antibody. In another example, an antibody molecule includes two heavy (H) chain variable domain sequences and two light (L) chain variable domain sequence, thereby forming two antigen binding sites, such as Fab, Fab’, F(ab’)2, Fc, Fd, Fd’, Fv, single chain antibodies (scFv for example), single variable domain antibodies, diabodies (Dab) (bivalent and bispecific), and chimeric (e.g., humanized) antibodies, which may be produced by the modification of whole antibodies or those synthesized de novo using recombinant DNA technologies. These functional antibody fragments retain the ability to selectively bind with their respective antigen or receptor. Antibodies and antibody fragments can be from any class of antibodies including, but not limited to, IgG, IgA, IgM, IgD, and IgE, and from any subclass (e.g., IgG1, IgG2, IgG3, and IgG4) of antibodies. A preparation of antibody molecules can be monoclonal or polyclonal. An antibody molecule can also be a human, humanized, CDR-grafted, or in vitro generated antibody. The antibody can have a heavy chain constant region chosen from, e.g., IgG1, IgG2, IgG3, or IgG4. The antibody can also have a light chain chosen from, e.g., kappa or lambda. The term “immunoglobulin” (Ig) is used interchangeably with the term “antibody” herein.

[0364] Examples of antigen-binding fragments of an antibody molecule include: (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab’)2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a diabody (dAb) fragment, which consists of a VH domain; (vi) a camelid or camelized variable domain; (vii) a single chain Fv (scFv), see e.g., Bird et al. (1988) Science 242:423-426; and Huston et al. (1988) Proc. Natl. Acad. Sci. USA 85:5879-5883); (viii) a singleWSGR Docket No.53676-767.601 domain antibody. These antibody fragments are obtained using conventional techniques known to those with skill in the art, and the fragments are screened for utility in the same manner as are intact antibodies.

[0365] Antibody molecules include intact molecules as well as functional fragments thereof. Constant regions of the antibody molecules can be altered, e.g., mutated, to modify the properties of the antibody (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function).

[0366] Antibody molecules can also be single domain antibodies. Single domain antibodies can include antibodies whose complementary determining regions are part of a single domain polypeptide. Examples include, but are not limited to, heavy chain antibodies, antibodies naturally devoid of light chains, single domain antibodies derived from conventional 4-chain antibodies, engineered antibodies and single domain scaffolds other than those derived from antibodies. Single domain antibodies may be any of the art, or any future single domain antibodies. Single domain antibodies may be derived from any species including, but not limited to mouse, human, camel, llama, fish, shark, goat, rabbit, and bovine. According to another aspect of the invention, a single domain antibody is a naturally occurring single domain antibody known as heavy chain antibody devoid of light chains. Such single domain antibodies are disclosed in WO 9404678, for example. For clarity reasons, this variable domain derived from a heavy chain antibody naturally devoid of light chain is known herein as a VHH or nanobody to distinguish it from the conventional VH of four chain immunoglobulins. Such a VHH molecule can be derived from antibodies raised in Camelidae species, for example in camel, llama, dromedary, alpaca and guanaco. Other species besides Camelidae may produce heavy chain antibodies naturally devoid of light chain; such VHHs are within the scope of the invention.

[0367] The VH and VL regions can be subdivided into regions of hypervariability, termed “complementarity determining regions” (CDR), interspersed with regions that are more conserved, termed “framework regions” (FR or FW).

[0368] The extent of the framework region and CDRs has been precisely defined by a number of methods (see, Kabat, E. A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.91-3242; Chothia, C. et al. (1987) J. Mol. Biol.196:901-917; and the AbM definition used by Oxford Molecular’s AbM antibody modeling software. See, generally, e.g., Protein Sequence and Structure Analysis of Antibody Variable Domains. In: Antibody Engineering Lab Manual (Ed.: Duebel, S. and Kontermann, R., Springer-Verlag, Heidelberg).

[0369] The terms “complementarity determining region,” and “CDR,” as used herein refer to the sequences of amino acids within antibody variable regions which confer antigen specificity and binding affinity. In general, there are three CDRs in each heavy chain variable region (HCDR1, HCDR2, HCDR3) and three CDRs in each light chain variable region (LCDR1, LCDR2, LCDR3).

[0370] The precise amino acid sequence boundaries of a given CDR can be determined using any of a number of known schemes, including those described by Kabat et al. (1991), “Sequences of Proteins of Immunological Interest,” 5thEd. Public Health Service, National Institutes of Health, Bethesda, MDWSGR Docket No.53676-767.601 (“Kabat” numbering scheme), Al-Lazikani et al., (1997) JMB 273,927-948 (“Chothia” numbering scheme), or by ImMunoGeneTics (IMGT) numbering system. As used herein, the CDRs defined according the “Chothia” number scheme are also sometimes referred to as “hypervariable loops.”

[0371] For example, under Kabat, the CDR amino acid residues in the heavy chain variable domain (VH) are numbered 31-35 (HCDR1), 50-65 (HCDR2), and 95-102 (HCDR3); and the CDR amino acid residues in the light chain variable domain (VL) are numbered 24-34 (LCDR1), 50-56 (LCDR2), and 89- 97 (LCDR3). Under Chothia, the CDR amino acids in the VH are numbered 26-32 (HCDR1), 52-56 (HCDR2), and 95-102 (HCDR3); and the amino acid residues in VL are numbered 26-32 (LCDR1), 50- 52 (LCDR2), and 91-96 (LCDR3).

[0372] Each VH and VL typically includes three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0373] The antibody molecule can be a polyclonal or a monoclonal antibody.

[0374] The terms “monoclonal antibody” or “monoclonal antibody composition” as used herein refer to a preparation of antibody molecules of single molecular composition. A monoclonal antibody composition displays a single binding specificity and affinity for a particular epitope. A monoclonal antibody can be made by hybridoma technology or by methods that do not use hybridoma technology (e.g., recombinant methods).

[0375] The antibody can be recombinantly produced, e.g., produced by phage display or by combinatorial methods.

[0376] Phage display and combinatorial methods for generating antibodies are known in the art (as described in, e.g., Ladner et al. U.S. Patent No. 5,223,409; Kang et al. International Publication No. WO 92 / 18619; Dower et al. International Publication No. WO 91 / 17271; Winter et al. International Publication WO 92 / 20791; Markland et al. International Publication No. WO 92 / 15679; Breitling et al. International Publication WO 93 / 01288; McCafferty et al. International Publication No. WO 92 / 01047; Garrard et al. International Publication No. WO 92 / 09690; Ladner et al. International Publication No. WO 90 / 02809; Fuchs et al. (1991) Bio / Technology 9:1370-1372; Hay et al. (1992) Hum Antibod Hybridomas 3:81-85; Huse et al. (1989) Science 246:1275-1281; Griffths et al. (1993) EMBO J 12:725- 734; Hawkins et al. (1992) J Mol Biol 226:889-896; Clackson et al. (1991) Nature 352:624-628; Gram et al. (1992) PNAS 89:3576-3580; Garrad et al. (1991) Bio / Technology 9:1373-1377; Hoogenboom et al. (1991) Nuc Acid Res 19:4133-4137; and Barbas et al. (1991) PNAS 88:7978-7982, the contents of all of which are incorporated by reference herein).

[0377] In one embodiment, the antibody is a fully human antibody (e.g., an antibody made in a mouse which has been genetically engineered to produce an antibody from a human immunoglobulin sequence), or a non-human antibody, e.g., a rodent (mouse or rat), goat, primate (e.g., monkey), camel antibody. Preferably, the non-human antibody is a rodent (mouse or rat antibody). Methods of producing rodent antibodies are known in the art.

[0378] Human monoclonal antibodies can be generated using transgenic mice carrying the human immunoglobulin genes rather than the mouse system. Splenocytes from these transgenic miceWSGR Docket No.53676-767.601 immunized with the antigen of interest are used to produce hybridomas that secrete human mAbs with specific affinities for epitopes from a human protein (see, e.g., Wood et al. International Application WO 91 / 00906, Kucherlapati et al. PCT publication WO 91 / 10741; Lonberg et al. International Application WO 92 / 03918; Kay et al. International Application 92 / 03917; Lonberg, N. et al.1994 Nature 368:856- 859; Green, L.L. et al.1994 Nature Genet.7:13-21; Morrison, S.L. et al.1994 Proc. Natl. Acad. Sci. USA 81:6851-6855; Bruggeman et al.1993 Year Immunol 7:33-40; Tuaillon et al.1993 PNAS 90:3720- 3724; Bruggeman et al.1991 Eur J Immunol 21:1323-1326).

[0379] An antibody molecule can be one in which the variable region, or a portion thereof, e.g., the CDRs, are generated in a non-human organism, e.g., a rat or mouse. Chimeric, CDR-grafted, and humanized antibodies are within the invention. Antibody molecules generated in a non-human organism, e.g., a rat or mouse, and then modified, e.g., in the variable framework or constant region, to decrease antigenicity in a human are within the invention.

[0380] An “effectively human” protein is a protein that does substantially not evoke a neutralizing antibody response, e.g., the human anti-murine antibody (HAMA) response. HAMA can be problematic in a number of circumstances, e.g., if the antibody molecule is administered repeatedly, e.g., in treatment of a chronic or recurrent disease condition. A HAMA response can make repeated antibody administration potentially ineffective because of an increased antibody clearance from the serum (see, e.g., Saleh et al., Cancer Immunol. Immunother., 32:180-190 (1990)) and also because of potential allergic reactions (see, e.g., LoBuglio et al., Hybridoma, 5:5117-5123 (1986)).

[0381] Chimeric antibodies can be produced by recombinant DNA techniques known in the art (see Robinson et al., International Patent Publication PCT / US86 / 02269; Akira, et al., European Patent Application 184,187; Taniguchi, M., European Patent Application 171,496; Morrison et al., European Patent Application 173,494; Neuberger et al., International Application WO 86 / 01533; Cabilly et al. U.S. Patent No.4,816,567; Cabilly et al., European Patent Application 125,023; Better et al. (1988 Science 240:1041-1043); Liu et al. (1987) PNAS 84:3439-3443; Liu et al., 1987, J. Immunol.139:3521-3526; Sun et al. (1987) PNAS 84:214-218; Nishimura et al., 1987, Canc. Res.47:999-1005; Wood et al. (1985) Nature 314:446-449; and Shaw et al., 1988, J. Natl Cancer Inst.80:1553-1559).

[0382] A humanized or CDR-grafted antibody will have at least one or two but generally all three recipient CDRs (of heavy and or light immuoglobulin chains) replaced with a donor CDR. The antibody may be replaced with at least a portion of a non-human CDR or only some of the CDRs may be replaced with non-human CDRs. It is only necessary to replace the number of CDRs required for binding to the antigen. Preferably, the donor will be a rodent antibody, e.g., a rat or mouse antibody, and the recipient will be a human framework or a human consensus framework. Typically, the immunoglobulin providing the CDRs is called the “donor” and the immunoglobulin providing the framework is called the “acceptor.” In one embodiment, the donor immunoglobulin is a non-human (e.g., rodent). The acceptor framework is a naturally occurring (e.g., a human) framework or a consensus framework, or a sequence about 85% or higher, preferably 90%, 95%, 99% or higher identical thereto.WSGR Docket No.53676-767.601

[0383] As used herein, the term “consensus sequence” refers to the sequence formed from the most frequently occurring amino acids (or nucleotides) in a family of related sequences (See e.g., Winnaker, From Genes to Clones (Verlagsgesellschaft, Weinheim, Germany 1987). In a family of proteins, each position in the consensus sequence is occupied by the amino acid occurring most frequently at that position in the family. If two amino acids occur equally frequently, either can be included in the consensus sequence. A “consensus framework” refers to the framework region in the consensus immunoglobulin sequence.

[0384] An antibody molecule can be humanized by methods known in the art (see e.g., Morrison, S. L., 1985, Science 229:1202-1207, by Oi et al., 1986, BioTechniques 4:214, and by Queen et al. US 5,585,089, US 5,693,761 and US 5,693,762, the contents of all of which are hereby incorporated by reference).

[0385] Humanized or CDR-grafted antibody molecules can be produced by CDR-grafting or CDR substitution, wherein one, two, or all CDRs of an immunoglobulin chain can be replaced. See, e.g., U.S. Patent 5,225,539; Jones et al.1986 Nature 321:552-525; Verhoeyan et al.1988 Science 239:1534; Beidler et al.1988 J. Immunol.141:4053-4060; Winter US 5,225,539, the contents of all of which are hereby expressly incorporated by reference. Winter describes a CDR-grafting method which may be used to prepare the humanized antibodies of the present invention (UK Patent Application GB 2188638A, filed on March 26, 1987; Winter US 5,225,539), the contents of which is expressly incorporated by reference.

[0386] Also within the scope of the invention are humanized antibody molecules in which specific amino acids have been substituted, deleted or added. Criteria for selecting amino acids from the donor are described in US 5,585,089, e.g., columns 12-16 of US 5,585,089, e.g., columns 12-16 of US 5,585,089, the contents of which are hereby incorporated by reference. Other techniques for humanizing antibodies are described in Padlan et al. EP 519596 A1, published on December 23, 1992.

[0387] The antibody molecule can be a single chain antibody. A single-chain antibody (scFv) may be engineered (see, for example, Colcher, D. et al. (1999) Ann N Y Acad Sci 880:263-80; and Reiter, Y. (1996) Clin Cancer Res 2:245-52). The single chain antibody can be dimerized or multimerized to generate multivalent antibodies having specificities for different epitopes of the same target protein.

[0388] In yet other embodiments, the antibody molecule has a heavy chain constant region chosen from, e.g., the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE; particularly, chosen from, e.g., the (e.g., human) heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4. In another embodiment, the antibody molecule has a light chain constant region chosen from, e.g., the (e.g., human) light chain constant regions of kappa or lambda. The constant region can be altered, e.g., mutated, to modify the properties of the antibody (e.g., to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, and / or complement function). In one embodiment the antibody has: effector function; and can fix complement. In other embodiments the antibody does not; recruit effector cells; or fix complement. In another embodiment, the antibody has reduced or no ability to bind an Fc receptor. For example, it is a isotype orWSGR Docket No.53676-767.601 subtype, fragment or other mutant, which does not support binding to an Fc receptor, e.g., it has a mutagenized or deleted Fc receptor binding region.

[0389] Methods for altering an antibody constant region are known in the art. Antibodies with altered function, e.g. altered affinity for an effector ligand, such as FcR on a cell, or the C1 component of complement can be produced by replacing at least one amino acid residue in the constant portion of the antibody with a different residue (see e.g., EP 388,151 A1, U.S. Pat. No.5,624,821 and U.S. Pat. No. 5,648,260, the contents of all of which are hereby incorporated by reference). Similar type of alterations could be described which if applied to the murine, or other species immunoglobulin would reduce or eliminate these functions.

[0390] An antibody molecule can be derivatized or linked to another functional molecule (e.g., another peptide or protein). As used herein, a “derivatized” antibody molecule is one that has been modified. Methods of derivatization include but are not limited to the addition of a fluorescent moiety, a radionucleotide, a toxin, an enzyme or an affinity ligand such as biotin. Accordingly, the antibody molecules of the invention are intended to include derivatized and otherwise modified forms of the antibodies described herein, including immunoadhesion molecules. For example, an antibody molecule can be functionally linked (by chemical coupling, genetic fusion, noncovalent association or otherwise) to one or more other molecular entities, such as another antibody (e.g., a bispecific antibody or a diabody), a detectable agent, a cytotoxic agent, a pharmaceutical agent, and / or a protein or peptide that can mediate association of the antibody or antibody portion with another molecule (such as a streptavidin core region or a polyhistidine tag).

[0391] One type of derivatized antibody molecule is produced by crosslinking two or more antibodies (of the same type or of different types, e.g., to create bispecific antibodies). Suitable crosslinkers include those that are heterobifunctional, having two distinctly reactive groups separated by an appropriate spacer (e.g., m-maleimidobenzoyl-N-hydroxysuccinimide ester) or homobifunctional (e.g., disuccinimidyl suberate). Such linkers are available from Pierce Chemical Company, Rockford, Ill. Multispecific or multifunctional antibody molecules

[0392] As used herein, a “multifunctional” or a “multispecific” molecule refers to molecule, e.g., a polypeptide, that has two or more functionalities, e.g., two or more binding specificities. In some embodiments, the functionalities can include one or more immune cell engagers, one or more tumor binding molecules, one or more cytokine molecules, one or more stromal modifiers, and other moieties described herein. In some embodiments, the multispecific molecule is a multispecific antibody molecule, e.g., a bispecific antibody molecule.

[0393] Exemplary structures of multispecific and multifunctional molecules defined herein are described throughout. Exemplary structures are further described in: Weidle U et al. (2013) The Intriguing Options of Multispecific Antibody Formats for Treatment of Cancer. Cancer Genomics & Proteomics 10: 1-18 (2013); and Spiess C et al. (2015) Alternative molecular formats and therapeutic applications forWSGR Docket No.53676-767.601 bispecific antibodies. Molecular Immunology 67: 95-106; the full contents of each of which is incorporated by reference herein).

[0394] In embodiments, multispecific antibody molecules can comprise more than one antigen-binding site, where different sites are specific for different antigens. In embodiments, multispecific antibody molecules can bind more than one (e.g., two or more) epitopes on the same antigen. In embodiments, multispecific antibody molecules comprise an antigen-binding site specific for a target cell (e.g., cancer cell) and a different antigen-binding site specific for an immune effector cell. In one embodiment, the multispecific antibody molecule is a bispecific antibody molecule. Bispecific antibody molecules can be classified into five different structural groups: (i) bispecific immunoglobulin G (BsIgG); (ii) IgG appended with an additional antigen-binding moiety; (iii) bispecific antibody fragments; (iv) bispecific fusion proteins; and (v) bispecific antibody conjugates.

[0395] BsIgG is a format that is monovalent for each antigen. Exemplary BsIgG formats include but are not limited to crossMab, DAF (two-in-one), DAF (four-in-one), DutaMab, DT-IgG, knobs-in-holes common LC, knobs-in-holes assembly, charge pair, Fab-arm exchange, SEEDbody, triomab, LUZ-Y, Fcab, ^ ^-body, orthogonal Fab. See Spiess et al. Mol. Immunol.67(2015):95-106. Exemplary BsIgGs include catumaxomab (Fresenius Biotech, Trion Pharma, Neopharm), which contains an anti-CD3 arm and an anti-EpCAM arm; and ertumaxomab (Neovii Biotech, Fresenius Biotech), which targets CD3 and HER2. In some embodiments, BsIgG comprises heavy chains that are engineered for heterodimerization. For example, heavy chains can be engineered for heterodimerization using a “knobs-into-holes” strategy, a SEED platform, a common heavy chain (e.g., in ^ ^-bodies), and use of heterodimeric Fc regions. See Spiess et al. Mol. Immunol.67(2015):95-106. Strategies that have been used to avoid heavy chain pairing of homodimers in BsIgG include knobs-in-holes, duobody, azymetric, charge pair, HA-TF, SEEDbody, and differential protein A affinity. See Id. BsIgG can be produced by separate expression of the component antibodies in different host cells and subsequent purification / assembly into a BsIgG. BsIgG can also be produced by expression of the component antibodies in a single host cell. BsIgG can be purified using affinity chromatography, e.g., using protein A and sequential pH elution.

[0396] IgG appended with an additional antigen-binding moiety is another format of bispecific antibody molecules. For example, monospecific IgG can be engineered to have bispecificity by appending an additional antigen-binding unit onto the monospecific IgG, e.g., at the N- or C- terminus of either the heavy or light chain. Exemplary additional antigen-binding units include single domain antibodies (e.g., variable heavy chain or variable light chain), engineered protein scaffolds, and paired antibody variable domains (e.g., single chain variable fragments or variable fragments). See Id. Examples of appended IgG formats include dual variable domain IgG (DVD-Ig), IgG(H)-scFv, scFv-(H)IgG, IgG(L)-scFv, scFv-(L)IgG, IgG(L,H)-Fv, IgG(H)-V, V(H)-IgG, IgG(L)-V, V(L)-IgG, KIH IgG-scFab, 2scFv-IgG, IgG-2scFv, scFv4-Ig, zybody, and DVI-IgG (four-in-one). See Spiess et al. Mol. Immunol.67(2015):95- 106. An example of an IgG-scFv is MM-141 (Merrimack Pharmaceuticals), which binds IGF-1R and HER3. Examples of DVD-Ig include ABT-981 (AbbVie), which binds IL-1α and IL-1β; and ABT-122 (AbbVie), which binds TNF and IL-17A.WSGR Docket No.53676-767.601

[0397] Bispecific antibody fragments (BsAb) are a format of bispecific antibody molecules that lack some or all of the antibody constant domains. For example, some BsAb lack an Fc region. In embodiments, bispecific antibody fragments include heavy and light chain regions that are connected by a peptide linker that permits efficient expression of the BsAb in a single host cell. Exemplary bispecific antibody fragments include but are not limited to nanobody, nanobody-HAS, BiTE, Diabody, DART, TandAb, scDiabody, scDiabody-CH3, Diabody-CH3, triple body, miniantibody, minibody, TriBi minibody, scFv-CH3 KIH, Fab-scFv, scFv-CH-CL-scFv, F(ab’)2, F(ab’)2-scFv2, scFv-KIH, Fab-scFv- Fc, tetravalent HCAb, scDiabody-Fc, Diabody-Fc, tandem scFv-Fc, and intrabody. See Id. For example, the BiTE format comprises tandem scFvs, where the component scFvs bind to CD3 on T cells and a surface antigen on cancer cells

[0398] Bispecific fusion proteins include antibody fragments linked to other proteins, e.g., to add additional specificity and / or functionality. An example of a bispecific fusion protein is an immTAC, which comprises an anti-CD3 scFv linked to an affinity-matured T-cell receptor that recognizes HLA- presented peptides. In embodiments, the dock-and-lock (DNL) method can be used to generate bispecific antibody molecules with higher valency. Also, fusions to albumin binding proteins or human serum albumin can be extend the serum half-life of antibody fragments. See Id.

[0399] In embodiments, chemical conjugation, e.g., chemical conjugation of antibodies and / or antibody fragments, can be used to create BsAb molecules. See Id. An exemplary bispecific antibody conjugate includes the CovX-body format, in which a low molecular weight drug is conjugated site-specifically to a single reactive lysine in each Fab arm or an antibody or fragment thereof. In embodiments, the conjugation improves the serum half-life of the low molecular weight drug. An exemplary CovX-body is CVX-241 (NCT01004822), which comprises an antibody conjugated to two short peptides inhibiting either VEGF or Ang2. See Id.

[0400] The antibody molecules can be produced by recombinant expression, e.g., of at least one or more component, in a host system. Exemplary host systems include eukaryotic cells (e.g., mammalian cells, e.g., CHO cells, or insect cells, e.g., SF9 or S2 cells) and prokaryotic cells (e.g., E. coli). Bispecific antibody molecules can be produced by separate expression of the components in different host cells and subsequent purification / assembly. Alternatively, the antibody molecules can be produced by expression of the components in a single host cell. Purification of bispecific antibody molecules can be performed by various methods such as affinity chromatography, e.g., using protein A and sequential pH elution. In other embodiments, affinity tags can be used for purification, e.g., histidine-containing tag, myc tag, or streptavidin tag. CDR-grafted scaffolds

[0401] In embodiments, the antibody molecule is a CDR-grafted scaffold domain. In embodiments, the scaffold domain is based on a fibronectin domain, e.g., fibronectin type III domain. The overall fold of the fibronectin type III (Fn3) domain is closely related to that of the smallest functional antibody fragment, the variable domain of the antibody heavy chain. There are three loops at the end of Fn3; theWSGR Docket No.53676-767.601 positions of BC, DE and FG loops approximately correspond to those of CDR1, 2 and 3 of the VH domain of an antibody. Fn3 does not have disulfide bonds; and therefore Fn3 is stable under reducing conditions, unlike antibodies and their fragments (see, e.g., WO 98 / 56915; WO 01 / 64942; WO 00 / 34784). An Fn3 domain can be modified (e.g., using CDRs or hypervariable loops described herein) or varied, e.g., to select domains that bind to an antigen / marker / cell described herein.

[0402] In embodiments, a scaffold domain, e.g., a folded domain, is based on an antibody, e.g., a “minibody” scaffold created by deleting three beta strands from a heavy chain variable domain of a monoclonal antibody (see, e.g., Tramontano et al., 1994, J Mol. Recognit.7:9; and Martin et al., 1994, EMBO J.13:5303-5309). The “minibody” can be used to present two hypervariable loops. In embodiments, the scaffold domain is a V-like domain (see, e.g., Coia et al. WO 99 / 45110) or a domain derived from tendamistatin, which is a 74 residue, six-strand beta sheet sandwich held together by two disulfide bonds (see, e.g., McConnell and Hoess, 1995, J Mol. Biol.250:460). For example, the loops of tendamistatin can be modified (e.g., using CDRs or hypervariable loops) or varied, e.g., to select domains that bind to a marker / antigen / cell described herein. Another exemplary scaffold domain is a beta- sandwich structure derived from the extracellular domain of CTLA-4 (see, e.g., WO 00 / 60070).

[0403] Other exemplary scaffold domains include but are not limited to T-cell receptors; MHC proteins; extracellular domains (e.g., fibronectin Type III repeats, EGF repeats); protease inhibitors (e.g., Kunitz domains, ecotin, BPTI, and so forth); TPR repeats; trifoil structures; zinc finger domains; DNA-binding proteins; particularly monomeric DNA binding proteins; RNA binding proteins; enzymes, e.g., proteases (particularly inactivated proteases), RNase; chaperones, e.g., thioredoxin, and heat shock proteins; and intracellular signaling domains (such as SH2 and SH3 domains). See, e.g., US 20040009530 and US 7,501,121, incorporated herein by reference.

[0404] In embodiments, a scaffold domain is evaluated and chosen, e.g., by one or more of the following criteria: (1) amino acid sequence, (2) sequences of several homologous domains, (3) 3-dimensional structure, and / or (4) stability data over a range of pH, temperature, salinity, organic solvent, oxidant concentration. In embodiments, the scaffold domain is a small, stable protein domain, e.g., a protein of less than 100, 70, 50, 40 or 30 amino acids. The domain may include one or more disulfide bonds or may chelate a metal, e.g., zinc. Antibody-Based Fusions

[0405] A variety of formats can be generated which contain additional binding entities attached to the N or C terminus of antibodies. These fusions with single chain or disulfide stabilized Fvs or Fabs result in the generation of tetravalent molecules with bivalent binding specificity for each antigen. Combinations of scFvs and scFabs with IgGs enable the production of molecules which can recognize three or more different antigens. Antibody-Fab Fusion

[0406] Antibody-Fab fusions are bispecific antibodies comprising a traditional antibody to a first target and a Fab to a second target fused to the C terminus of the antibody heavy chain. Commonly the antibodyWSGR Docket No.53676-767.601 and the Fab will have a common light chain. Antibody fusions can be produced by (1) engineering the DNA sequence of the target fusion, and (2) transfecting the target DNA into a suitable host cell to express the fusion protein. It seems like the antibody-scFv fusion may be linked by a (Gly)-Ser linker between the C-terminus of the CH3 domain and the N-terminus of the scFv, as described by Coloma, J. et al. (1997) Nature Biotech 15:159. Antibody-scFv Fusion

[0407] Antibody-scFv Fusions are bispecific antibodies comprising a traditional antibody and a scFv of unique specificity fused to the C terminus of the antibody heavy chain. The scFv can be fused to the C terminus through the Heavy Chain of the scFv either directly or through a linker peptide. Antibody fusions can be produced by (1) engineering the DNA sequence of the target fusion, and (2) transfecting the target DNA into a suitable host cell to express the fusion protein. It seems like the antibody-scFv fusion may be linked by a (Gly)-Ser linker between the C-terminus of the CH3 domain and the N- terminus of the scFv, as described by Coloma, J. et al. (1997) Nature Biotech 15:159. Variable Domain Immunoglobulin DVD

[0408] A related format is the dual variable domain immunoglobulin (DVD), which are composed of VH and VL domains of a second specificity place upon the N termini of the V domains by shorter linker sequences.

[0409] Other exemplary multispecific antibody formats include, e.g., those described in the following US20160114057A1, US20130243775A1, US20140051833, US20130022601, US20150017187A1, US20120201746A1, US20150133638A1, US20130266568A1, US20160145340A1, WO2015127158A1, US20150203591A1, US20140322221A1, US20130303396A1, US20110293613, US20130017200A1, US20160102135A1, WO2015197598A2, WO2015197582A1, US9359437, US20150018529, WO2016115274A1, WO2016087416A1, US20080069820A1, US9145588B, US7919257, and US20150232560A1. Exemplary multispecific molecules utilizing a full antibody-Fab / scFab format include those described in the following, US9382323B2, US20140072581A1, US20140308285A1, US20130165638A1, US20130267686A1, US20140377269A1, US7741446B2, and WO1995009917A1. Exemplary multispecific molecules utilizing a domain exchange format include those described in the following, US20150315296A1, WO2016087650A1, US20160075785A1, WO2016016299A1, US20160130347A1, US20150166670, US8703132B2, US20100316645, US8227577B2, US20130078249. Fc-containing entities (mini-antibodies)

[0410] Fc-containing entities, also known as mini-antibodies, can be generated by fusing scFv to the C- termini of constant heavy region domain 3 (CH3-scFv) and / or to the hinge region (scFv-hinge-Fc) of an antibody with a different specificity. Trivalent entities can also be made which have disulfide stabilized variable domains (without peptide linker) fused to the C-terminus of CH3 domains of IgGs.WSGR Docket No.53676-767.601 Fc-containing multispecific molecules

[0411] In some embodiments, the multispecific molecules disclosed herein includes an immunoglobulin constant region (e.g., an Fc region). Exemplary Fc regions can be chosen from the heavy chain constant regions of IgG1, IgG2, IgG3 or IgG4; more particularly, the heavy chain constant region of human IgG1, IgG2, IgG3, or IgG4.

[0412] In some embodiments, the immunoglobulin chain constant region (e.g., the Fc region) is altered, e.g., mutated, to increase or decrease one or more of: Fc receptor binding, antibody glycosylation, the number of cysteine residues, effector cell function, or complement function.

[0413] In other embodiments, an interface of a first and second immunoglobulin chain constant regions (e.g., a first and a second Fc region) is altered, e.g., mutated, to increase or decrease dimerization, e.g., relative to a non-engineered interface, e.g., a naturally occurring interface. For example, dimerization of the immunoglobulin chain constant region (e.g., the Fc region) can be enhanced by providing an Fc interface of a first and a second Fc region with one or more of: a paired protuberance-cavity (“knob-in-a hole”), an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms, e.g., relative to a non-engineered interface.

[0414] In some embodiments, the multispecific molecules include a paired amino acid substitution at a position chosen from one or more of 347, 349, 350, 351, 366, 368, 370, 392, 394, 395, 397, 398, 399, 405, 407, or 409, e.g., of the Fc region of human IgG1 For example, the immunoglobulin chain constant region (e.g., Fc region) can include a paired an amino acid substitution chosen from: T366S, L368A, or Y407V (e.g., corresponding to a cavity or hole), and T366W (e.g., corresponding to a protuberance or knob).

[0415] In other embodiments, the multifunctional molecule includes a half-life extender, e.g., a human serum albumin or an antibody molecule to human serum albumin. Heterodimerized Antibody Molecules & Methods of Making

[0416] Various methods of producing multispecific antibodies have been disclosed to address the problem of incorrect heavy chain pairing. Exemplary methods are described below. Exemplary multispecific antibody formats and methods of making said multispecific antibodies are also disclosed in e.g., Speiss et al. Molecular Immunology 67 (2015) 95–106; and Klein et al mAbs 4:6, 653–663; November / December 2012; the entire contents of each of which are incorporated by reference herein.

[0417] Heterodimerized bispecific antibodies are based on the natural IgG structure, wherein the two binding arms recognize different antigens. IgG derived formats that enable defined monovalent (and simultaneous) antigen binding are generated by forced heavy chain heterodimerization, combined with technologies that minimize light chain mispairing (e.g., common light chain). Forced heavy chain heterodimerization can be obtained using, e.g., knob-in-hole OR strand exchange engineered domains (SEED).WSGR Docket No.53676-767.601 Knob-in-Hole

[0418] Knob-in-Hole as described in US 5,731,116, US 7,476,724 and Ridgway, J. et al. (1996) Prot. Engineering 9(7): 617-621, broadly involves: (1) mutating the CH3 domain of one or both antibodies to promote heterodimerization; and (2) combining the mutated antibodies under conditions that promote heterodimerization. “Knobs” or “protuberances” are typically created by replacing a small amino acid in a parental antibody with a larger amino acid (e.g., T366Y or T366W); “Holes” or “cavities” are created by replacing a larger residue in a parental antibody with a smaller amino acid (e.g., Y407T, T366S, L368A and / or Y407V).

[0419] For bispecific antibodies including an Fc domain, introduction of specific mutations into the constant region of the heavy chains to promote the correct heterodimerization of the Fc portion can be utilized. Several such techniques are reviewed in Klein et al. (mAbs (2012) 4:6, 1-11), the contents of which are incorporated herein by reference in their entirety. These techniques include the “knobs-into- holes” (KiH) approach which involves the introduction of a bulky residue into one of the CH3 domains of one of the antibody heavy chains. This bulky residue fits into a complementary “hole” in the other CH3 domain of the paired heavy chain so as to promote correct pairing of heavy chains (see e.g., US7642228).

[0420] Exemplary KiH mutations include S354C, T366W in the “knob” heavy chain and Y349C, T366S, L368A, Y407V in the “hole” heavy chain. Other exemplary KiH mutations are provided in Table 1, with additional optional stabilizing Fc cysteine mutations.

[0421] Other Fc mutations are provided by Igawa and Tsunoda who identified 3 negatively charged residues in the CH3 domain of one chain that pair with three positively charged residues in the CH3 domain of the other chain. These specific charged residue pairs are: E356-K439, E357-K370, D399- K409 and vice versa. By introducing at least two of the following three mutations in chain A: E356K, E357K and D399K, as well as K370E, K409D, K439E in chain B, alone or in combination with newly identified disulfide bridges, they were able to favor very efficient heterodimerization while suppressing homodimerization at the same time (Martens T et al. A novel one-armed antic- Met antibody inhibits glioblastoma growth in vivo. Clin Cancer Res 2006; 12:6144-52; PMID:17062691). Xencor defined 41 variant pairs based on combining structural calculations and sequence information that were subsequently screened for maximal heterodimerization, defining the combination of S364H, F405A (HA) on chain A and Y349T, T394F on chain B (TF) (Moore GL et al. A novel bispecific antibody format enables simultaneous bivalent and monovalent co-engagement of distinct target antigens. MAbs 2011; 3:546-57; PMID: 22123055).

[0422] Other exemplary Fc mutations to promote heterodimerization of multispecific antibodies include those described in the following references, the contents of each of which is incorporated by reference herein, WO2016071377A1, US20140079689A1, US20160194389A1, US20160257763, WO2016071376A2, WO2015107026A1, WO2015107025A1, WO2015107015A1, US20150353636A1, US20140199294A1, US7750128B2, US20160229915A1, US20150344570A1, US8003774A1, US20150337049A1, US20150175707A1, US20140242075A1, US20130195849A1, US20120149876A1,WSGR Docket No.53676-767.601 US20140200331A1, US9309311B2, US8586713, US20140037621A1, US20130178605A1, US20140363426A1, US20140051835A1 and US20110054151A1.

[0423] Stabilizing cysteine mutations have also been used in combination with KiH and other Fc heterodimerization promoting variants, see e.g., US7183076. Other exemplary cysteine modifications include, e.g., those disclosed in US20140348839A1, US7855275B2, and US9000130B2. Strand Exchange Engineered Domains (SEED)

[0424] Heterodimeric Fc platform that support the design of bispecific and asymmetric fusion proteins by devising strand-exchange engineered domain (SEED) C(H)3 heterodimers are known. These derivatives of human IgG and IgA C(H)3 domains create complementary human SEED C(H)3 heterodimers that are composed of alternating segments of human IgA and IgG C(H)3 sequences. The resulting pair of SEED C(H)3 domains preferentially associates to form heterodimers when expressed in mammalian cells. SEEDbody (Sb) fusion proteins consist of [IgG1 hinge]-C(H)2-[SEED C(H)3], that may be genetically linked to one or more fusion partners (see e.g., Davis JH et al. SEEDbodies: fusion proteins based on strand exchange engineered domain (SEED) CH3 heterodimers in an Fc analogue platform for asymmetric binders or immunofusions and bispecific antibodies. Protein Eng Des Sel 2010; 23:195-202; PMID:20299542 and US8871912. The contents of each of which are incorporated by reference herein). Duobody

[0425] “Duobody” technology to produce bispecific antibodies with correct heavy chain pairing are known. The DuoBody technology involves three basic steps to generate stable bispecific human IgG1antibodies in a post-production exchange reaction. In a first step, two IgG1s, each containing single matched mutations in the third constant (CH3) domain, are produced separately using standard mammalian recombinant cell lines. Subsequently, these IgG1 antibodies are purified according to standard processes for recovery and purification. After production and purification (post-production), the two antibodies are recombined under tailored laboratory conditions resulting in a bispecific antibody product with a very high yield (typically >95%) (see e.g., Labrijn et al, PNAS 2013;110(13):5145-5150 and Labrijn et al. Nature Protocols 2014;9(10):2450-63, the contents of each of which are incorporated by reference herein). Electrostatic Interactions

[0426] Methods of making multispecific antibodies using CH3 amino acid changes with charged amino acids such that homodimer formation is electrostatically unfavorable are disclosed. EP1870459 and WO 2009089004 describe other strategies for favoring heterodimer formation upon co-expression of different antibody domains in a host cell. In these methods, one or more residues that make up the heavy chain constant domain 3 (CH3), CH3-CH3 interfaces in both CH3 domains are replaced with a charged amino acid such that homodimer formation is electrostatically unfavorable and heterodimerization is electrostatically favorable. Additional methods of making multispecific molecules using electrostatic interactions are described in the following references, the contents of each of which is incorporated byWSGR Docket No.53676-767.601 reference herein, include US20100015133, US8592562B2, US9200060B2, US20140154254A1, and US9358286A1. Common Light Chain

[0427] Light chain mispairing needs to be avoided to generate homogenous preparations of bispecific IgGs. One way to achieve this is through the use of the common light chain principle, i.e. combining two binders that share one light chain but still have separate specificities. An exemplary method of enhancing the formation of a desired bispecific antibody from a mixture of monomers is by providing a common variable light chain to interact with each of the heteromeric variable heavy chain regions of the bispecific antibody. Compositions and methods of producing bispecific antibodies with a common light chain as disclosed in, e.g., US7183076B2, US20110177073A1, EP2847231A1, WO2016079081A1, and EP3055329A1, the contents of each of which is incorporated by reference herein. Common Heavy Chain

[0428] An exemplary method of enhancing the formation of a desired bispecific antibody from a mixture of monomers is by providing a common variable heavy chain to interact with each of the heteromeric variable light chain regions of the bispecific antibody. Compositions and methods of producing bispecific antibodies with a common heavy chain are disclosed in, e.g., US20120184716, US20130317200, and US20160264685A1, the contents of each of which is incorporated by reference herein. CrossMab

[0429] Another option to reduce light chain mispairing is the CrossMab technology which avoids non- specific L chain mispairing by exchanging CH1 and CL domains in the Fab of one half of the bispecific antibody. Such crossover variants retain binding specificity and affinity, but make the two arms so different that L chain mispairing is prevented. The CrossMab technology (as reviewed in Klein et al. Supra) involves domain swapping between heavy and light chains so as to promote the formation of the correct pairings. Briefly, to construct a bispecific IgG-like CrossMab antibody that could bind to two antigens by using two distinct light chain–heavy chain pairs, a two-step modification process is applied. First, a dimerization interface is engineered into the C-terminus of each heavy chain using a heterodimerization approach, e.g., Knob-into-hole (KiH) technology, to ensure that only a heterodimer of two distinct heavy chains from one antibody (e.g., Antibody A) and a second antibody (e.g., Antibody B) is efficiently formed. Next, the constant heavy 1 (CH1) and constant light (CL) domains of one antibody are exchanged (Antibody A), keeping the variable heavy (VH) and variable light (VL) domains consistent. The exchange of the CH1 and CL domains ensured that the modified antibody (Antibody A) light chain would only efficiently dimerize with the modified antibody (antibody A) heavy chain, while the unmodified antibody (Antibody B) light chain would only efficiently dimerize with the unmodified antibody (Antibody B) heavy chain; and thus only the desired bispecific CrossMab would be efficiently formed (see e.g., Cain, C. SciBX 4(28); doi:10.1038 / scibx.2011.783, the contents of which are incorporated by reference herein).WSGR Docket No.53676-767.601 Amino Acid Modifications

[0430] Alternative compositions and methods of producing multispecific antibodies with correct light chain pairing include various amino acid modifications. For example, Zymeworks describes heterodimers with one or more amino acid modifications in the CH1 and / or CL domains, one or more amino acid modifications in the VH and / or VL domains, or a combination thereof, which are part of the interface between the light chain and heavy chain and create preferential pairing between each heavy chain and a desired light chain such that when the two heavy chains and two light chains of the heterodimer pair are co-expressed in a cell, the heavy chain of the first heterodimer preferentially pairs with one of the light chains rather than the other (see e.g., WO2015181805). Other exemplary methods are described in WO2016026943 (Argen-X), US20150211001, US20140072581A1, US20160039947A1, and US20150368352. Lambda / Kappa Formats

[0431] Multispecific molecules (e.g., multispecific antibody molecules) that include the lambda light chain polypeptide and a kappa light chain polypeptide, can be used to allow for heterodimerization. Methods for generating bispecific antibody molecules comprising the lambda light chain polypeptide and a kappa light chain polypeptide are disclosed in PCT / US17 / 53053 filed on September 22, 2017, incorporated herein by reference in its entirety.

[0432] In embodiments, the multispecific molecules includes a multispecific antibody molecule, e.g., an antibody molecule comprising two binding specificities, e.g., a bispecific antibody molecule. The multispecific antibody molecule includes: a lambda light chain polypeptide 1 (LLCP1) specific for a first epitope; a heavy chain polypeptide 1 (HCP1) specific for the first epitope; a kappa light chain polypeptide 2 (KLCP2) specific for a second epitope; and a heavy chain polypeptide 2 (HCP2) specific for the second epitope.

[0433] “Lambda light chain polypeptide 1 (LLCP1)”, as that term is used herein, refers to a polypeptide comprising sufficient light chain (LC) sequence, such that when combined with a cognate heavy chain variable region, can mediate specific binding to its epitope and complex with an HCP1. In an embodiment it comprises all or a fragment of a CH1 region. In an embodiment, an LLCP1 comprises LC-CDR1, LC-CDR2, LC-CDR3, FR1, FR2, FR3, FR4, and CH1, or sufficient sequence therefrom to mediate specific binding of its epitope and complex with an HCP1. LLCP1, together with its HCP1, provide specificity for a first epitope (while KLCP2, together with its HCP2, provide specificity for a second epitope). As described elsewhere herein, LLCP1 has a higher affinity for HCP1 than for HCP2.

[0434] “Kappa light chain polypeptide 2 (KLCP2)”, as that term is used herein, refers to a polypeptide comprising sufficient light chain (LC) sequence, such that when combined with a cognate heavy chain variable region, can mediate specific binding to its epitope and complex with an HCP2. In an embodiment, it comprises all or a fragment of a CH1 region. In an embodiment, a KLCP2 comprises LC-CDR1, LC-CDR2, LC-CDR3, FR1, FR2, FR3, FR4, and CH1, or sufficient sequence therefrom toWSGR Docket No.53676-767.601 mediate specific binding of its epitope and complex with an HCP2. KLCP2, together with its HCP2, provide specificity for a second epitope (while LLCP1, together with its HCP1, provide specificity for a first epitope).

[0435] “Heavy chain polypeptide 1 (HCP1)”, as that term is used herein, refers to a polypeptide comprising sufficient heavy chain (HC) sequence, e.g., HC variable region sequence, such that when combined with a cognate LLCP1, can mediate specific binding to its epitope and complex with an HCP1. In an embodiment, it comprises all or a fragment of a CH1region. In an embodiment, it comprises all or a fragment of a CH2 and / or CH3 region. In an embodiment an HCP1 comprises HC-CDR1, HC-CDR2, HC-CDR3, FR1, FR2, FR3, FR4, CH1, CH2, and CH3, or sufficient sequence therefrom to: (i) mediate specific binding of its epitope and complex with an LLCP1, (ii) to complex preferentially, as described herein to LLCP1 as opposed to KLCP2; and (iii) to complex preferentially, as described herein, to an HCP2, as opposed to another molecule of HCP1. HCP1, together with its LLCP1, provide specificity for a first epitope (while KLCP2, together with its HCP2, provide specificity for a second epitope).

[0436] “Heavy chain polypeptide 2 (HCP2)”, as that term is used herein, refers to a polypeptide comprising sufficient heavy chain (HC) sequence, e.g., HC variable region sequence, such that when combined with a cognate LLCP1, can mediate specific binding to its epitope and complex with an HCP1. In an embodiment, it comprises all or a fragment of a CH1region. In an embodiment, it comprises all or a fragment of a CH2 and / or CH3 region. In an embodiment an HCP1 comprises HC-CDR1, HC-CDR2, HC-CDR3, FR1, FR2, FR3, FR4, CH1, CH2, and CH3, or sufficient sequence therefrom to: (i) mediate specific binding of its epitope and complex with an KLCP2, (ii) to complex preferentially, as described herein to KLCP2 as opposed to LLCP1; and (iii) to complex preferentially, as described herein, to an HCP1, as opposed to another molecule of HCP2. HCP2, together with its KLCP2, provide specificity for a second epitope (while LLCP1, together with its HCP1, provide specificity for a first epitope).

[0437] In some embodiments of the multispecific antibody molecule disclosed herein: LLCP1 has a higher affinity for HCP1 than for HCP2; and / or KLCP2 has a higher affinity for HCP2 than for HCP1.

[0438] In embodiments, the affinity of LLCP1 for HCP1 is sufficiently greater than its affinity for HCP2, such that under preselected conditions, e.g., in aqueous buffer, e.g., at pH 7, in saline, e.g., at pH 7, or under physiological conditions, at least 75, 80, 90, 95, 98, 99, 99.5, or 99.9 % of the multispecific antibody molecule molecules have a LLCP1complexed, or interfaced with, a HCP1.

[0439] In some embodiments of the multispecific antibody molecule disclosed herein: the HCP1 has a greater affinity for HCP2, than for a second molecule of HCP1; and / or the HCP2 has a greater affinity for HCP1, than for a second molecule of HCP2.

[0440] In embodiments, the affinity of HCP1 for HCP2 is sufficiently greater than its affinity for a second molecule of HCP1, such that under preselected conditions, e.g., in aqueous buffer, e.g., at pH 7, in saline, e.g., at pH 7, or under physiological conditions, at least 75%, 80, 90, 95, 98, 9999.5 or 99.9 % of the multispecific antibody molecule molecules have a HCP1complexed, or interfaced with, a HCP2.WSGR Docket No.53676-767.601

[0441] In another aspect, disclosed herein is a method for making, or producing, a multispecific antibody molecule. The method includes: (i) providing a first heavy chain polypeptide (e.g., a heavy chain polypeptide comprising one, two, three or all of a first heavy chain variable region (first VH), a first CH1, a first heavy chain constant region (e.g., a first CH2, a first CH3, or both)); (ii) providing a second heavy chain polypeptide (e.g., a heavy chain polypeptide comprising one, two, three or all of a second heavy chain variable region (second VH), a second CH1, a second heavy chain constant region (e.g., a second CH2, a second CH3, or both)); (iii) providing a lambda chain polypeptide (e.g., a lambda light variable region (VLλ), a lambda light constant chain (VLλ), or both) that preferentially associates with the first heavy chain polypeptide (e.g., the first VH); and (iv) providing a kappa chain polypeptide (e.g., a kappa light variable region (VLκ), a kappa light constant chain (VLκ), or both) that preferentially associates with the second heavy chain polypeptide (e.g., the second VH), under conditions where (i)-(iv) associate.

[0442] In embodiments, the first and second heavy chain polypeptides form an Fc interface that enhances heterodimerization.

[0443] In embodiments, (i)-(iv) (e.g., nucleic acid encoding (i)-(iv)) are introduced in a single cell, e.g., a single mammalian cell, e.g., a CHO cell. In embodiments, (i)-(iv) are expressed in the cell.

[0444] In embodiments, (i)-(iv) (e.g., nucleic acid encoding (i)-(iv)) are introduced in different cells, e.g., different mammalian cells, e.g., two or more CHO cell. In embodiments, (i)-(iv) are expressed in the cells.

[0445] In one embodiment, the method further comprises purifying a cell-expressed antibody molecule, e.g., using a lambda- and / or- kappa-specific purification, e.g., affinity chromatography.

[0446] In embodiments, the method further comprises evaluating the cell-expressed multispecific antibody molecule. For example, the purified cell-expressed multispecific antibody molecule can be analyzed by techniques known in the art, include mass spectrometry. In one embodiment, the purified cell-expressed antibody molecule is cleaved, e.g., digested with papain to yield the Fab moieties and evaluated using mass spectrometry.

[0447] In embodiments, the method produces correctly paired kappa / lambda multispecific, e.g., bispecific, antibody molecules in a high yield, e.g., at least 75%, 80, 90, 95, 98, 9999.5 or 99.9 %.

[0448] In other embodiments, the multispecific, e.g., a bispecific, antibody molecule that includes: (i) a first heavy chain polypeptide (HCP1) (e.g., a heavy chain polypeptide comprising one, two, three or all of a first heavy chain variable region (first VH), a first CH1, a first heavy chain constant region (e.g., a first CH2, a first CH3, or both)), e.g., wherein the HCP1 binds to a first epitope; (ii) a second heavy chain polypeptide (HCP2) (e.g., a heavy chain polypeptide comprising one, two, three or all of a second heavy chain variable region (second VH), a second CH1, a second heavy chain constant region (e.g., a second CH2, a second CH3, or both)), e.g., wherein the HCP2 binds to a second epitope;WSGR Docket No.53676-767.601 (iii) a lambda light chain polypeptide (LLCP1) (e.g., a lambda light variable region (VLλ), a lambda light constant chain (VLλ), or both) that preferentially associates with the first heavy chain polypeptide (e.g., the first VH), e.g., wherein the LLCP1 binds to a first epitope; and (iv) a kappa light chain polypeptide (KLCP2) (e.g., a kappa light variable region (VLκ), a kappa light constant chain (VLκ), or both) that preferentially associates with the second heavy chain polypeptide (e.g., the second VH), e.g., wherein the KLCP2 binds to a second epitope.

[0449] In embodiments, the first and second heavy chain polypeptides form an Fc interface that enhances heterodimerization. In embodiments, the multispecific antibody molecule has a first binding specificity that includes a hybrid VLλ-CLλ heterodimerized to a first heavy chain variable region connected to the Fc constant, CH2-CH3 domain (having a knob modification) and a second binding specificity that includes a hybrid VLκ-CLκ heterodimerized to a second heavy chain variable region connected to the Fc constant, CH2-CH3 domain (having a hole modification). TRBC1 and TRBC2 Antigen Binding Domains

[0450] The present disclosure provides, inter alia, antibody molecules, e.g., multispecific (e.g., bi-, tri-, tetra- specific) or multifunctional molecules, that include, e.g., are engineered to contain, one or more antigen binding domains that bind to a tumor antigen on a lymphoma cell (e.g., T cell). In some embodiments, the tumor antigen comprises a T cell receptor comprising TRBC1 or TRBC2. In some embodiments, the antigen binding domain preferentially binds to a T cell receptor comprising TRBC1 (e.g., relative to a T cell receptor comprising TRBC2). In some embodiments, the antigen binding domain preferentially binds to a T cell receptor comprising TRBC2 (e.g., relative to a T cell receptor comprising TRBC1). In some embodiments, the multifunctional molecules include, e.g., are engineered to contain, one or more antigen binding domains that selectively target lymphocytes expressing TRBC1 or TRBC2. In some embodiments, the antigen binding domain selectively targets lymphocytes expressing a T cell receptor comprising TRBC1 or a T cell receptor comprising TRBC2.

[0451] T cell receptors (TCRs) are receptors found on the surface of lymphocytes, specifically on T lymphocytes (T cells). TCRs are responsible for recognizing antigen fragments presented by major histocompatibility complex (MHC) molecules on other immune cells (e.g., B cells) by signaling through associated CD3 and activating the T cell. The vast majority of TCRs in humans are heterodimers comprising an alpha chain and a beta chain. Both alpha and beta chains of TCR comprise variable and constant regions. The variable regions of the alpha and beta chain are encoded by distinct DNA elements (V, D, and J elements for beta chain; V and J elements for the alpha chain). Recombination between these elements produces in large part the variation in antigen binding specificity of TCRs. The TCR beta chain constant region is selected from two different domains, beta constant domain 1 and beta constant domain 2. Without wishing to be bound by theory, it is thought that the majority of TCRs comprising a beta chain comprise a beta chain comprising beta constant domain 1 or beta constant domain 2, but not both constant domain 1 and constant domain 2.WSGR Docket No.53676-767.601

[0452] In some embodiments, the multifunctional or multispecific molecules or antibody molecules of the present application comprise an antigen binding domain that binds to a tumor antigen on a lymphoma cell (e.g., a T cell), e.g., a T cell receptor comprising TRBC1, TRBC1, a T cell receptor comprising TRBC2, or TRBC2. In some embodiments, the multifunctional or multispecific molecules or antibody molecules of the present application comprise an antigen binding domain that selectively targets lymphocytes expressing a T cell receptor comprising TRBC1, TRBC1, a T cell receptor comprising TRBC2, or TRBC2. While it is most typical for a lymphocyte or lymphoma cell presenting a T cell receptor comprising TRBC1 or TRBC2 to be a T cell, cancer causes many disruptions in non-disease expression patterns. Thus, in some embodiments, the lymphoma cell or lymphocyte may not be a T cell. In some embodiments, the lymphoma cell or lymphocyte is a B cell. In some embodiments, the lymphoma cell or lymphocyte is a natural killer cell.

[0453] In some embodiments, the antigen binding domain (e.g., first antigen binding domain) comprises any CDR amino acid sequence, framework region (FWR) amino acid sequence, or variable region amino acid sequence of an anti-TRBC1 antibody known in the art. In some embodiments, CDR amino acid sequence, framework region (FWR) amino acid sequence, or variable region amino acid sequence are selected from JOVI.1. TRBC1 Antigen Binding Domains

[0454] In some embodiments, the antigen binding domain that binds to TRBC1 comprises one or more CDRs (e.g., VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and / or VLCDR3) disclosed in Table 2A or Table 2B, Table 3A or Table 3B, or Table 4, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC1 comprises one or more framework regions (e.g., VHFWR1, VHFWR2, VHFWR3, VHFWR4, VLFWR1, VLFWR2, VLFWR3, and / or VLFWR4) disclosed in Table 2A or Table 2B,Table 3A or Table 3B, or Table 4, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC1 comprises a VH and / or a VL disclosed in Table 7, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC1 comprises an amino acid sequence disclosed in Table 8, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto.

[0455] In some embodiments, the antigen binding domain that binds to TRBC1 comprises one or more CDRs (e.g., VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and / or VLCDR3) disclosed in Table 5A and / or 5B, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC1 comprises one or more framework regions (e.g., VHFWR1, VHFWR2, VHFWR3, VHFWR4, VLFWR1, VLFWR2, VLFWR3, and / or VLFWR4) disclosed in Table 5A and / or 5B, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC1 comprises a VH and / or a VL disclosed in Table 7, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto.WSGR Docket No.53676-767.601

[0456] In some embodiments, the antigen binding domain that binds to TRBC1 comprises a VH comprising a heavy chain complementarity determining region 1 (VHCDR1), a VHCDR2, and a VHCDR3, and a VL comprising a light chain complementarity determining region 1 (VLCDR1), a VLCDR2, and a VLCDR3.

[0457] In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 8643, 7355, and 202, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 8643, 201, and 202, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7354, 201, and 202, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7354, 7355, and 202, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0458] In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 223, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7367, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 223, 7368, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 223, 224, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7367, 7368, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0459] In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 8643, 7355, 202, 223, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 8643, 201, 202, 223, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of: SEQ ID NOs: 8643, 7355, 202, 7367, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 7355, 202, 223, 7368, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 7355, 202, 223, 224, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 7355, 202, 7367, 7368, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 202, 7367, 224, and 225, respectively (or a sequenceWSGR Docket No.53676-767.601 having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 202, 223, 7368, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 202, 223, 224, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 202, 7367, 7368, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 201, 202, 223, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 201, 202, 7367, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 201, 202, 223, 7368, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 201, 202, 223, 224, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 201, 202, 7367, 7368, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 7355, 202, 223, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 7355, 202, 7367, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 7355, 202, 223, 7368, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7354, 7355, 202, 223, 224, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); or SEQ ID NOs: 7354, 7355, 202, 7367, 7368, and 7369, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0460] In some embodiments, the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 7351, 253, 250-252, 254, 7343, 7344, 7350, and 7352 (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto) and / or the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 258, 255-257, 259, 260, and 7357-7360 (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7351 and 258, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 253 and 258, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0461] In some embodiments, the antigen binding domain (e.g., first antigen binding domain) that binds to a tumor antigen on a lymphoma cell (e.g., a T cell), e.g., a T cell receptor comprising TRBC1, TRBC1, a T cell receptor comprising TRBC2, or TRBC2 comprises any CDR amino acid sequence, framework region (FWR) amino acid sequence, or variable region amino acid sequence Table 1, Table 2A or Table 2B,Table 3A or Table 3B, Table 4, Table 7, and Table 8. In some embodiments, the antigen binding domain (e.g., first antigen binding domain) that binds to a tumor antigen on a lymphoma cell (e.g., a T cell), e.g., a T cell receptor comprising TRBC1, TRBC1, a T cell receptor comprising TRBC2, or TRBC2 comprises heavy and / or light chain amino acid sequences of Table 8. In some embodiments, the antigen binding domain (e.g., first antigen binding domain) that selectively targets lymphocytes expressing a T cell receptor comprising TRBC1, TRBC1, a T cell receptor comprising TRBC2, or TRBC2 comprises any CDR amino acid sequence, framework region (FWR) amino acid sequence, or variable region aminoWSGR Docket No.53676-767.601 acid sequence disclosed in Table 1, Table 2A or Table 2B,Table 3A or Table 3B, Table 4, Table 7, and Table 8. In some embodiments, the antigen binding domain (e.g., first antigen binding domain) that selectively targets lymphocytes expressing a T cell receptor comprising TRBC1, TRBC1, a T cell receptor comprising TRBC2, or TRBC2 comprises heavy and / or light chain amino acid sequences of Table 8. An antigen binding domain that binds to a tumor antigen comprising TRBC1 or selectively targets lymphocytes expressing TRBC1 may be said to target TRBC1 (i.e., a TRBC1-targeting antigen binding domain). An antigen binding domain that binds to a tumor antigen comprising TRBC2 or selectively targets lymphocytes expressing TRBC2 may be said to target TRBC2 (i.e., a TRBC2- targeting antigen binding domain).

[0462] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a heavy chain complementarity determining region 1 (VHCDR1) amino acid sequence of SEQ ID NO: 8643 (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), a VHCDR2 amino acid sequence of SEQ ID NO: 201 (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), and / or a VHCDR3 amino acid sequence of SEQ ID NO: 202 (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions). In some embodiments, the TRBC1 antigen binding domain comprises a VH comprising a VHCDR1 amino acid sequence of SEQ ID NO: 8643, a VHCDR2 amino acid sequence of SEQ ID NO: 201, and / or a VHCDR3 amino acid sequence of SEQ ID NO: 202.

[0463] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain complementarity determining region 1 (VLCDR1) amino acid sequence of SEQ ID NO: 223 (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), a VLCDR2 amino acid sequence of SEQ ID NO: 224 (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions), and / or a VLCDR3 amino acid sequence of SEQ ID NO: 225 (or a sequence with no more than 1, 2, 3, or 4 mutations, e.g., substitutions, additions, or deletions). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLCDR1 amino acid sequence of SEQ ID NO: 223, a VLCDR2 amino acid sequence of SEQ ID NO: 224, and a VLCDR3 amino acid sequence of SEQ ID NO: 225.

[0464] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a heavy chain framework region 1 (VHFWR1) amino acid sequence of SEQ ID NO: 203, a VHFWR2 amino acid sequence of SEQ ID NO: 204, a VHFWR3 amino acid sequence of SEQ ID NO: 205, and / or a VHFWR4 amino acid sequence of SEQ ID NO: 206.

[0465] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain framework region 1 (VLFWR1) amino acid sequence of SEQ ID NO: 226, a VLFWR2 amino acid sequence of SEQ ID NO: 227, a VLFWR3 amino acid sequence of SEQ ID NO: 228, and / or a VLFWR4 amino acid sequence of SEQ ID NO: 229.

[0466] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a VHFWR1 amino acid sequence of SEQ ID NO: 203 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR2 amino acidWSGR Docket No.53676-767.601 sequence of SEQ ID NO: 204 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR3 amino acid sequence of SEQ ID NO: 205 (or a sequence with no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 mutations, e.g., substitutions, additions, or deletions), and / or a VHFWR4 amino acid sequence of SEQ ID NO: 206.

[0467] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLFWR1 amino acid sequence of SEQ ID NO: 226 (or a sequence with no more than 1, 2, or 3 mutations, e.g., substitutions, additions, or deletions), a VLFWR2 amino acid sequence of SEQ ID NO: 227 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), a VLFWR3 amino acid sequence of SEQ ID NO: 228 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), and / or a VLFWR4 amino acid sequence of SEQ ID NO: 229.

[0468] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a heavy chain framework region 1 (VHFWR1) amino acid sequence of SEQ ID NO: 207, a VHFWR2 amino acid sequence of SEQ ID NO: 208, a VHFWR3 amino acid sequence of SEQ ID NO: 209, and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0469] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a VHFWR1 amino acid sequence of SEQ ID NO: 207 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR2 amino acid sequence of SEQ ID NO: 208 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR3 amino acid sequence of SEQ ID NO: 209 (or a sequence with no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 mutations, e.g., substitutions, additions, or deletions), and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0470] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a heavy chain framework region 1 (VHFWR1) amino acid sequence of SEQ ID NO: 207, a VHFWR2 amino acid sequence of SEQ ID NO: 208, a VHFWR3 amino acid sequence of SEQ ID NO: 213, and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0471] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a VHFWR1 amino acid sequence of SEQ ID NO: 207 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR2 amino acid sequence of SEQ ID NO: 208 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR3 amino acid sequence of SEQ ID NO: 213 (or a sequence with no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 mutations, e.g., substitutions, additions, or deletions), and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0472] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a heavy chain framework region 1 (VHFWR1) amino acid sequence of SEQ ID NO: 7345, a VHFWR2 amino acid sequence of SEQ ID NO: 208, a VHFWR3 amino acid sequence of SEQ ID NO: 217, and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0473] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a VHFWR1 amino acid sequence of SEQ ID NO: 7345 (or a sequence with no more than 1,WSGR Docket No.53676-767.601 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR2 amino acid sequence of SEQ ID NO: 208 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR3 amino acid sequence of SEQ ID NO: 217 (or a sequence with no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 mutations, e.g., substitutions, additions, or deletions), and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0474] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a heavy chain framework region 1 (VHFWR1) amino acid sequence of SEQ ID NO: 207, a VHFWR2 amino acid sequence of SEQ ID NO: 212, a VHFWR3 amino acid sequence of SEQ ID NO: 221, and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0475] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising a VHFWR1 amino acid sequence of SEQ ID NO: 207 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR2 amino acid sequence of SEQ ID NO: 212 (or a sequence with no more than 1, 2, 3, 4, 5, or 6 mutations, e.g., substitutions, additions, or deletions, therefrom), a VHFWR3 amino acid sequence of SEQ ID NO: 221 (or a sequence with no more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 mutations, e.g., substitutions, additions, or deletions), and / or a VHFWR4 amino acid sequence of SEQ ID NO: 210.

[0476] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain framework region 1 (VLFWR1) amino acid sequence of SEQ ID NO: 230, a VLFWR2 amino acid sequence of SEQ ID NO: 231, a VLFWR3 amino acid sequence of SEQ ID NO: 232, and / or a VLFWR4 amino acid sequence of SEQ ID NO: 307.

[0477] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLFWR1 amino acid sequence of SEQ ID NO: 230 (or a sequence with no more than 1, 2, or 3 mutations, e.g., substitutions, additions, or deletions), a VLFWR2 amino acid sequence of SEQ ID NO: 231 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), a VLFWR3 amino acid sequence of SEQ ID NO: 232 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), and / or a VLFWR4 amino acid sequence of SEQ ID NO: 307.

[0478] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain framework region 1 (VLFWR1) amino acid sequence of SEQ ID NO: 234, a VLFWR2 amino acid sequence of SEQ ID NO: 235, a VLFWR3 amino acid sequence of SEQ ID NO: 236, and / or a VLFWR4 amino acid sequence of SEQ ID NO: 237.

[0479] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLFWR1 amino acid sequence of SEQ ID NO: 234 (or a sequence with no more than 1, 2, or 3 mutations, e.g., substitutions, additions, or deletions), a VLFWR2 amino acid sequence of SEQ ID NO: 235 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), a VLFWR3 amino acid sequence of SEQ ID NO: 236 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), and / or a VLFWR4 amino acid sequence of SEQ ID NO: 237.

[0480] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain framework region 1 (VLFWR1) amino acid sequence of SEQ ID NO: 238, aWSGR Docket No.53676-767.601 VLFWR2 amino acid sequence of SEQ ID NO: 239, a VLFWR3 amino acid sequence of SEQ ID NO: 240, and / or a VLFWR4 amino acid sequence of SEQ ID NO: 241.

[0481] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLFWR1 amino acid sequence of SEQ ID NO: 238 (or a sequence with no more than 1, 2, or 3 mutations, e.g., substitutions, additions, or deletions), a VLFWR2 amino acid sequence of SEQ ID NO: 239 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), a VLFWR3 amino acid sequence of SEQ ID NO: 240 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), and / or a VLFWR4 amino acid sequence of SEQ ID NO: 241.

[0482] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain framework region 1 (VLFWR1) amino acid sequence of SEQ ID NO: 242, a VLFWR2 amino acid sequence of SEQ ID NO: 243, a VLFWR3 amino acid sequence of SEQ ID NO: 240, and / or a VLFWR4 amino acid sequence of SEQ ID NO: 245.

[0483] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLFWR1 amino acid sequence of SEQ ID NO: 242 (or a sequence with no more than 1, 2, or 3 mutations, e.g., substitutions, additions, or deletions), a VLFWR2 amino acid sequence of SEQ ID NO: 243 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), a VLFWR3 amino acid sequence of SEQ ID NO: 240 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), and / or a VLFWR4 amino acid sequence of SEQ ID NO: 245.

[0484] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a light chain framework region 1 (VLFWR1) amino acid sequence of SEQ ID NO: 246, a VLFWR2 amino acid sequence of SEQ ID NO: 247, a VLFWR3 amino acid sequence of SEQ ID NO: 240, and / or a VLFWR4 amino acid sequence of SEQ ID NO: 249.

[0485] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising a VLFWR1 amino acid sequence of SEQ ID NO: 246 (or a sequence with no more than 1, 2, or 3 mutations, e.g., substitutions, additions, or deletions), a VLFWR2 amino acid sequence of SEQ ID NO: 247 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), a VLFWR3 amino acid sequence of SEQ ID NO: 240 (or a sequence with no more than 1 mutation, e.g., substitution, addition, or deletion), and / or a VLFWR4 amino acid sequence of SEQ ID NO: 249.

[0486] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 250 (or an amino acid sequence having at least about 77%, 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 250). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 255 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 255). In some embodiments, antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 250. In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 255.WSGR Docket No.53676-767.601

[0487] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 250, and a VL comprising the amino acid sequence of SEQ ID NO: 255.

[0488] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 251 (or an amino acid sequence having at least about 77%, 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 251). In some embodiments, antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 251.

[0489] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 252 (or an amino acid sequence having at least about 77%, 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 252). In some embodiments, antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 252.

[0490] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 253 (or an amino acid sequence having at least about 77%, 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 253). In some embodiments, antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 253.

[0491] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 254 (or an amino acid sequence having at least about 77%, 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 254). In some embodiments, antigen binding domain that targets TRBC1 comprises a VH comprising the amino acid sequence of SEQ ID NO: 254.

[0492] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 256 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 256). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 256.

[0493] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 257 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 257). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 257.

[0494] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 258 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 258). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 258.WSGR Docket No.53676-767.601

[0495] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 259 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 259). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 259.

[0496] In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 260 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 260). In some embodiments, the antigen binding domain that targets TRBC1 comprises a VL comprising the amino acid sequence of SEQ ID NO: 260.

[0497] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6154 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6154). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6154.

[0498] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4671 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 4671). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4671.

[0499] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6155 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6155). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6155.

[0500] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4672 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 4672). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4672.

[0501] In some embodiments, the antigen binding domain that targets TRBC1 comprises a light chain comprising the amino acid sequence of SEQ ID NO: 6156 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6156). In some embodiments, the antigen binding domain that targets TRBC1 comprises a light chain comprising the amino acid sequence of SEQ ID NO: 6156.

[0502] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6167 (or an amino acid sequence having at leastWSGR Docket No.53676-767.601 about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6167). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6167.

[0503] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4673 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 4673). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4673.

[0504] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6168 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6168). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6168.

[0505] n some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4674 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 4674). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 4674.

[0506] In some embodiments, the antigen binding domain that targets TRBC1 comprises a light chain comprising the amino acid sequence of SEQ ID NO: 6169 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6169). In some embodiments, the antigen binding domain that targets TRBC1 comprises a light chain comprising the amino acid sequence of SEQ ID NO: 6169.

[0507] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6154 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6154) and a light chain comprising the amino acid sequence of SEQ ID NO: 6156 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6156). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6154 and a light chain comprising the amino acid sequence of SEQ ID NO: 6156.

[0508] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6155 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6155) and a light chain comprising the amino acid sequence of SEQ ID NO: 6156 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6156). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6155 and a light chain comprising the amino acid sequence of SEQ ID NO: 6156.WSGR Docket No.53676-767.601

[0509] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6167 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6167) and a light chain comprising the amino acid sequence of SEQ ID NO: 6169 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6169). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6167 and a light chain comprising the amino acid sequence of SEQ ID NO: 6169.

[0510] In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6168 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6168) and a light chain comprising the amino acid sequence of SEQ ID NO: 6169 (or an amino acid sequence having at least about 93%, 95%, or 99% sequence identity to SEQ ID NO: 6169). In some embodiments, the antigen binding domain that targets TRBC1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 6168 and a light chain comprising the amino acid sequence of SEQ ID NO: 6169. TRBC2 Antigen Binding Domains

[0511] In some embodiments, the antigen binding domain that binds to TRBC2 comprises one or more CDRs (e.g., VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and / or VLCDR3) disclosed in Table 9A, Table 9B, or Table 10, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC2 comprises one or more framework regions (e.g., VHFWR1, VHFWR2, VHFWR3, VHFWR4, VLFWR1, VLFWR2, VLFWR3, and / or VLFWR4) disclosed in Table 9A, Table 9B, or Table 10, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC2 comprises a VH and / or a VL disclosed in Table 11, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto. In some embodiments, the antigen binding domain that binds to TRBC2 comprises an amino acid sequence disclosed in Table 12, or a sequence having at least 85%, 90%, 95%, or 99% identity thereto.

[0512] In some embodiments, the antigen binding domain that binds to TRBC2 comprises a VH comprising a heavy chain complementarity determining region 1 (VHCDR1), a VHCDR2, and a VHCDR3, and a VL comprising a light chain complementarity determining region 1 (VLCDR1), a VLCDR2, and a VLCDR3.

[0513] In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7441, 201, and 7442, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7422, 201, and 7403, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7401, 201, and 7403, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1,WSGR Docket No.53676-767.601 VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7394, 201, and 307, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 8643, 201, and 7398, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 8643, 201, and 7400, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7405, 201, and 7403, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7407, 201, and 7403, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7427, 201, and 7403, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, and VHCDR3 comprise the amino acid sequences of SEQ ID NOs: 7430, 201, and 7403, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0514] In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7443, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0515] In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7441, 201, 7442, 7443, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7422, 201, 7403, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of SEQ ID NOs: 7401, 201, 7403, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VHCDR1, VHCDR2, VHCDR3, VLCDR1, VLCDR2, and VLCDR3 comprise the amino acid sequences of: SEQ ID NOs: 7394, 201, 307, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 7398, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 7400, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7405, 201, 7403, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs:WSGR Docket No.53676-767.601 7407, 201, 7403, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7427, 201, 7403, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7430, 201, 7403, 7410, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7422, 201, 7403, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7401, 201, 7403, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7394, 201, 307, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 7398, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 8643, 201, 7400, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7405, 201, 7403, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7407, 201, 7403, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); SEQ ID NOs: 7427, 201, 7403, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto); or SEQ ID NOs: 7430, 201, 7403, 7409, 224, and 225, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0516] In some embodiments, the VH comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 7420, 7423, 7411, 7412, 7413, 7414, 7415, 7416, 7417, 7425, 7428, and 7431 (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto) and / or the VL comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 7419 and 7418 (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7420 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7423 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7411 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7412 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7413 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7414 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7415 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7416 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7417 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In someWSGR Docket No.53676-767.601 embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7425 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7428 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7431 and 7419, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7420 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7423 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7411 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7412 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7413 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7414 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7415 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7416 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7417 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7425 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7428 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto). In some embodiments, the VH and VL comprise the amino acid sequences of SEQ ID NOs: 7431 and 7418, respectively (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0517] In some embodiments, the antigen binding domain that binds to TRBC2 comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 7433, 7434, 7435, 7436, and 7437 (or a sequence having at least 85%, 90%, 95%, or 99% identity thereto).

[0518] In some embodiments, a bispecific antibody is contemplated herein to comprise a first antigen binding domain that binds to TRBC2. In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain and a light chain of a TRBC2 binding antibody (e.g., TRCBC2 binder-1, or TRCBC2 binder-2 or TRCBC2 binder 3, or TRCBC2 binder 4). In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain variable domain selected from SEQ ID NOs: 8011,WSGR Docket No.53676-767.601 8013, 8020, or 8303, or a sequence that is at least 90% identical to SEQ ID NOs: 8011, 8013, 8020, or 8303; and a light chain variable domain selected from SEQ ID NOs: 8012, 8014, 8014, or 8014 or a sequence that is at least 90% identical to SEQ ID NOs: 1, 3, 31, or 33, including any combinations thereof.

[0519] In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain and a light chain of TRBC2 binding antibody e.g., TRCBC2 binder-1. In some embodiments, an exemplary TRBC2 binding domain comprises a sequence of amino acids in the heavy chain e.g., a heavy chain variable domain (VH) that is at least 90% identical to SEQ ID NO: 8011. In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain variable domain that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8011. In some embodiments, the TRBC2 binding domain comprises a heavy chain comprising an amino acid sequence of SEQ ID NO: 8011.

[0520] In some embodiments, an exemplary TRBC2 binding domain comprises a sequence of amino acids in the light chain e.g., a light chain variable domain (VL) that is at least 95% identical to SEQ ID NO: 8012. In some embodiments, the light chain variable domain is at least at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8012. In some embodiments, the TRBC2 binding domain comprises a light chain comprising an amino acid sequence of SEQ ID NO: 8012. In some embodiments, the bispecific antibody comprises a first antigen binding domain that binds to TRBC2 and comprises a heavy chain VH domain (SEQ ID NO: 8011) and a light chain VL domain (SEQ ID NO: 8012) of a TRBC2 binder, or a TRBC2 binder having one or more amino acid sequences described in Table 13 or Table 14 , or a TRBC2 binder having any one of the domains described in Table 13 or Table 14 , or one or more domains with one or more amino acid modifications relative to the amino acid sequences of domains described in Table 13 or Table 14.

[0521] In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain and a light chain of TRBC2 binding antibody e.g., TRCBC2 binder-2. In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain that is at least 95% identical to SEQ ID NO:8013. In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain that is at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8013. In some embodiments, an exemplary TRBC2 binding domain comprises a light chain that is at least 95% identical to SEQ ID NO: 8014. In some embodiments, an exemplary TRBC2 binding domain comprises a light chain that is at least at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8014. In some embodiments, the bispecific antibody comprises a first antigen binding domain that binds to TRBC2 that comprises a heavy chain VH (SEQ ID NO: 8013) and a light chain VL (SEQ ID NO: 8014) of a TRBC2 binder, or a TRBC2 binder having any one of the domains described in Table 13, or one or more domains with one or more amino acid modifications relative to the amino acid sequences of domains described in Table 13.

[0522] In some embodiments, an exemplary TRBC2 binding domain comprises a sequence of amino acids in the heavy chain e.g., a heavy chain variable domain (VH) that is at least 90% identical to SEQWSGR Docket No.53676-767.601 ID NO: 8020. In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain variable domain that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8020.

[0523] In some embodiments, an exemplary TRBC2 binding domain comprises a sequence of amino acids in the light chain e.g., a light chain variable domain (VL) that is at least 90% identical to SEQ ID NO: 8014. In some embodiments, an exemplary TRBC2 binding domain comprises a light chain variable domain that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8014. In some embodiments, the bispecific antibody comprises a first antigen binding domain that binds to TRBC2 and comprises a heavy chain VH (SEQ ID NO: 8020) and a light chain VL (SEQ ID NO: 8014) of a TRBC2 binder, or a TRBC2 binder having any one of the domains described in Table 13, or one or more domains with one or more amino acid modifications relative to the amino acid sequences of domains described in Table 13 or Table 14.

[0524] In some embodiments, an exemplary TRBC2 binding domain comprises a sequence of amino acids in the heavy chain e.g., a heavy chain variable domain (VH) that is at least 90% identical to SEQ ID NO: 8303. In some embodiments, an exemplary TRBC2 binding domain comprises a heavy chain variable domain that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to SEQ ID NO: 8303.

[0525] In some embodiments, an exemplary TRBC2 binding domain comprises a sequence of amino acids in the light chain e.g., a light chain variable domain (VL) that is at least 90% identical to SEQ ID NO: 8014. In some embodiments, the bispecific antibody comprises a first antigen binding domain that binds to TRBC2 and comprises a heavy chain VH (SEQ ID NO: 8013) and a light chain VL (SEQ ID NO: 8024) of a TRBC2 binder, or a TRBC2 binder having any one of the domains described in Table 13, or one or more domains with one or more amino acid modifications relative to the amino acid sequences of domains described in Table 13.

[0526] For example, a TRBC2 -binding Heavy Chain domain may comprise the sequence of SEQ ID NO:8011 QVQLVQSGAEVKKPGSSVKVSCKASPRGFYGYHMHWVRQAPGQGLEWMGFINPYTNDIQYNE RFRGRVTITSDESTTTAYMELSSLRSEDTAVYYCAMGNGKWGDGAYRFFDLWGQGTLVTVSS (SEQ ID NO: 8011 ), and a TRBC2 -binding Light Chain domain may comprise the sequence of SEQ ID NO:8012 DVVMTQSPLSLPVTLGQPASISCRSSENLVHSNGRTYLQWYQQRPGQSPRLLIYRVSNRFPGVPD RFSGSGSGTDFTLKISRVEAEDVGVYFCSQSSLEPYTFGGGTKVEIK (SEQ ID NO: 8012).

[0527] For example, a TRBC2 -binding Heavy Chain domain may comprise the sequence of SEQ ID NO:8013 QVQLVQSGAEVKKPGSSVKVSCKASPRGFYGYHMHWVRQAPGQGLEWMGFINPYNNHIQYNE RFRGRVTITSDESTTTAYMELSSLRSEDTAVYYCALGNGKWGDGAYRFFDFWGQGTLVTVSS (SEQ ID NO: 8013), and a TRBC2 -binding Light Chain domain may comprise the sequence of SEQ ID NO: 8014WSGR Docket No.53676-767.601 DVVMTQSPLSLPVTLGQPASISCRSSKNLVHSNGRTYLQWYQQRPGQSPRLLIYRVSNRFPGVPD RFSGSGSGTDFTLKISRVEAEDVGVYFCSQSTREPYTFGGGTKVEIK (SEQ ID NO: 8014)

[0528] A TRBC2 -binding Heavy Chain domain may comprise the sequence of SEQ ID NO:8020 QVQLVQSGAEVKKPGSSVKVSCKASPRGFYGYHMHWVRQAPGQGLEWMGFINPYNNHIQYNE RFRGRVTITSDESTTTAYMELSSLRSEDTAVYYCALGEGKWGDGAYRFFDFWGQGTLVTVSS (SEQ ID NO: 8020), and a TRBC2 -binding Light Chain domain may comprise the sequence of SEQ ID NO: 8014: DVVMTQSPLSLPVTLGQPASISCRSSKNLVHSNGRTYLQWYQQRPGQSPRLLIYRVSNRFPGVPD RFSGSGSGTDFTLKISRVEAEDVGVYFCSQSTREPYTFGGGTKVEIK (SEQ ID NO: 8014).

[0529] A TRBC2 -binding Heavy Chain domain may comprise the sequence of SEQ ID NO:8303 QVQLVQSGAEVKKPGSSVKVSCKASPRGFYGYHMHWVRQAPGQGLEWMGFINPYNNHIQYNE RFRGRVTITSDESTTTAYMELSSLRSEDTAVYYCALGAGKWGDGAYRFFDFWGQGTLVTVSS (SEQ ID NO: 8303), and a TRBC2 -binding Light Chain domain may comprise the sequence of SEQ ID NO: 8014: DVVMTQSPLSLPVTLGQPASISCRSSKNLVHSNGRTYLQWYQQRPGQSPRLLIYRVSNRFPGVPD RFSGSGSGTDFTLKISRVEAEDVGVYFCSQSTREPYTFGGGTKVEIK (SEQ ID NO: 8014).

[0530] In some embodiments, the TRBC2 antigen binding domain may comprise a mutation in the heavy chain variable domain, the mutation may be selected from a T28K mutation, a Y32F mutation and an A100N mutation in the VH domain compared to a VH domain of an antibody as described in SEQ ID NO: 8024 In some embodiments, the TRBC2 antigen binding domain comprises two or more mutations in the heavy chain variable domain, the mutation may be selected from a T28K mutation, a Y32F mutation and an A100N mutation in the VH domain compared to VH domain of the antibody as described in SEQ ID NO: 8024. In some embodiments, the VH domain may comprise a mutation that may be at position V2, or Y27, or G31 or R98, or Y102, or N103 or A107 with respect to SEQ ID NO: 8024. In some embodiments, the VH domain of the TRBC2 antigen binding domain may comprise a V2K or a V2R mutation compared to VH domain of the antibody as described in SEQ ID NO: 8024. In some embodiments, the VH domain of the TRBC2 antigen binding domain may comprise a Y27F, Y27M, Y27N or Y27W mutation compared to VH domain of the antibody as described in SEQ ID NO: 8024. In some embodiments, the VH domain of the TRBC2 antigen binding domain may comprise a G31K, a G31R, or a G31S mutation compared to VH domain of the antibody as described in SEQ ID NO: 8024. In some embodiments, the VH domain of the TRBC2 antigen binding domain may comprise a R98K mutation compared to VH domain of the antibody as described in SEQ ID NO: 8024. In some embodiments, the VH domain of the TRBC2 antigen binding domain may comprise a Y102F or Y102L mutation compared to VH domain of the antibody as described in SEQ ID NO: 8024. In some embodiments, the VH domain of the TRBC2 antigen binding domain may comprise a N193A, N193E, N103F, N103H, N103L, N103M, N103Q, N103S, N103W, or N103Y mutation compared to VH domain of the antibody as described in SEQ ID NO: 8024.WSGR Docket No.53676-767.601

[0531] In some embodiments, the TRBC2 antigen binding domain may comprise a mutation in the light chain variable domain, the mutation may be at position N35, or at R55 with respect to the VL domain of an antibody as described in SEQ ID NO: 8025. In some embodiments, the VL domain of the TRBC2 antigen binding domain may comprise a N35M, N35F, N35Y, N35K or N35R mutation at position 35 with respect to the VL domain of an antibody as described in SEQ ID NO: 8025.

[0532] SEQ ID NO: 8024 – Reference antibody VH domain: QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYVMHWVRQAPGQGLEWMGFINPYNDDIQSNE RFRGRVTMTRDT SISTAYMELSRLRSDDTAVYYCARGAGYNFDGAYRFFDFWGQGTMVTVSS (SEQ ID NO: 8024).

[0533] SEQ ID NO: 8025 – Reference antibody VL domain: DIVMTQSPLSLPVTPGEPASISCRSSQRLVHSNGNTYLHWYLQKPGQSPRLLIYRVSNRFPGVPDR FSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPYTFGQGTKLEIK (SEQ ID NO: 8025).

[0534] In some embodiments, the bispecific antibody comprises a second antigen binding domain that binds to NKp30. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain and a light chain of NKp30 binder 1. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain having an amino acid sequence of SEQ ID NO: 8006. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain having an amino acid sequence that is at least 90% identical to a sequence of SEQ ID NO: 8006. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain having an amino acid sequence that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a sequence of SEQ ID NO: 8006.

[0535] In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain and a light chain of NKp30 binder 1. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain having an amino acid sequence of SEQ ID NO: 4684. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain having an amino acid sequence that is at least 90% identical to a sequence of SEQ ID NO: 4684. In some embodiments, an exemplary NKp30 binding domain comprises a heavy chain having an amino acid sequence that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a sequence of SEQ ID NO: 4684.

[0536] In some embodiments, an exemplary NKp30 binding domain comprises a light chain of SEQ ID NO: 8003. In some embodiments, an exemplary NKp30 binding domain comprises a light chain of SEQ ID NO: 4683. In some embodiments, an exemplary NKp30 binding domain comprises a light chain having an amino acid sequence that is at least 90% identical to a sequence of SEQ ID NO: 8003. In some embodiments, an exemplary NKp30 binding domain comprises a light chain having an amino acid sequence that is at least 90% identical to a sequence of SEQ ID NO: 4683. In some embodiments, an exemplary NKp30 binding domain comprises a light chain having an amino acid sequence that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a sequence of SEQ ID NO: 8003. In some embodiments, an exemplary NKp30WSGR Docket No.53676-767.601 binding domain comprises a light chain having an amino acid sequence that is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a sequence of SEQ ID NO: 4683. In some embodiments, the bispecific antibody comprising a second antigen binding domain that binds to NKp30 comprises a heavy chain and a light chain of NKp30 binder 1, having amino acid sequences described in below.

[0537] An exemplary NKp30-binding, Heavy Chain domain may comprise the sequence of SEQ ID NO: 8030: EIQLLESGGGLVQPGGSLRLSCAVSGFSITTTGYHWNWVRQAPGKGLEWVGYIYSSGSTSYNPS LKSRFTISRDTSKNTFYLQMNSLRAEDTAVYYCARGDWHYFDYWGQGTMVTVSS (SEQ ID NO: 8030).

[0538] An exemplary NKp30-binding, Light Chain domain may comprise the sequence of SEQ ID NO: 13: DSVTTQSPLSLPVTLGQPASISCSGEKLSDKYVHWYQQRPGQSPRMLIYENDRRPSGVPDRFSGS NSGNDATLKISRVEAEDVGVYFCQFWDSTNSAVFGGGTKVEIK (SEQ ID NO: 13)

[0539] In some embodiments, an exemplary bispecific antibody comprises (i) a first antigen binding domain comprising a TRBC2 binding domain having a heavy chain sequence of SEQ ID NO: 8011, or a sequence that is at least 90% identical to SEQ ID NO: 8011; and a light chain sequence of SEQ ID NO: 8012, or a sequence that is at least 95% identical to SEQ ID NO: 8012; and (ii) a second antigen binding domain comprising a NKp30 binding domain having a heavy chain and a light chain sequence of SEQ ID NOs: 8030 and 13 respectively.

[0540] In some embodiments, an exemplary bispecific antibody comprises (i) a first antigen binding domain comprising a TRBC2 binding domain having a heavy chain sequence of SEQ ID NO: 8013, or a sequence that is at least 95% identical to SEQ ID NO: 8013; and a light chain sequence of SEQ ID NO: 8014, or a sequence that is at least 95% identical to SEQ ID NO: 8014; and (ii) a second antigen binding domain comprising a NKp30 binding domain having a heavy chain and a light chain sequence of SEQ ID NO: 8030 and 13 respectively.

[0541] In some embodiments, an exemplary bispecific antibody comprises (i) a first antigen binding domain comprising a TRBC2 binding domain having a heavy chain sequence of SEQ ID NO: 8011, or a sequence that is at least 90% identical to SEQ ID NO: 8011 and a light chain sequence of SEQ ID NO: 8012, or a sequence that is at least 95% identical to SEQ ID NO: 8012; and (ii) a second antigen binding domain comprising a NKp30 binding domain having a heavy chain and a light chain sequence at least 90% identical to SEQ ID NO: 8030 and at least 90% identical to SEQ ID NO: 13 respectively.

[0542] In some embodiments, an exemplary bispecific antibody comprises (i) a first antigen binding domain comprising a TRBC2 binding domain having a heavy chain sequence of SEQ ID NO: 8013, or a sequence that is at least 95% identical to SEQ ID NO: 8013 and a light chain sequence of SEQ ID NO: 8014, or a sequence that is at least 95% identical to SEQ ID NO: 8014; and (ii) a second antigen binding domain comprising a NKp30 binding domain having a heavy chain and a light chain sequence at least 90% identical to SEQ ID NO: 8030 and at least 90% identical to SEQ ID NO: 13 respectively.WSGR Docket No.53676-767.601

[0543] In some embodiments, the bi-specific antibody comprising : (i) a first antigen binding domain comprising a TRBC2 binding domain that comprises a single chain variable fragment (scFv); wherein the scFv comprises a heavy chain and a light chain that are linked by a short linker, and (ii) a second antigen binding domain comprising a NKp30 binding domain that comprises a single chain variable fragment (scFv); wherein scFv comprises a heavy chain and a light chain that are linked by a short linker.

[0544] In some embodiments, the heavy chain of a TRBC2 binding domain comprises a HC CDR1, a CDR2 and a CDR3. In some embodiments, the light chain of a TRBC2 binding domain comprises a LC CDR1, a CDR2 and a CDR3. In one embodiment, the heavy chain of a TRBC2 binding domain comprises a HC CDR1 having an amino acid sequence comprising PRGFYGYHMH (SEQ ID NO: 8272) {or PRGFYGY (SEQ ID NO: 8041 according to Chothia numbering system)}, or PRGFYGYAMH (SEQ ID NO: 8272). In one embodiment, the heavy chain of a TRBC2 binding domain comprises a HC CDR1 having an amino acid sequence RSGFHGYAMH (SEQ ID NO: 8207) or PRGFHGYHMH (SEQ ID NO: 8211). In some embodiments, the heavy chain TRBC2 binding domain comprises a HC CDR1 comprising an amino acid sequence SRSGFHGYAMH (SEQ ID NO: 8215). In some embodiments, the heavy chain TRBC2 binding domain comprises a HC CDR1 comprising an amino acid sequence PRGFYGYHMH (SEQ ID NO: 8272). In some embodiments, the heavy chain TRBC2 binding domain comprises a HC CDR1 comprising an amino acid sequence RSSQNLVHSNGRTYLH (SEQ ID NO: 8226). In one embodiment, the heavy chain of a TRBC2 binding domain comprises a HC CDR2 having an amino acid sequence MGFINPYTNDIQYNERFRG (SEQ ID NO: 8042). In one embodiment, the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GNGKWGDGAYRFFDL (SEQ ID NO: 8043). In one embodiment, the heavy chain of a TRBC2 binding domain comprises a HC CDR2 comprising an amino acid sequence FINPYNNHIQYNERFRG (SEQ ID NO: 8044) or FINPYNDDIQYNQKFQG (SEQ ID NO: 8208) or YINPYNRDIKYNQKFQG (SEQ ID NO: 8212). In some embodiments, the HC CDR2 has an amino acid sequence FINPYNHAIKYNQKFQG (SEQ ID NO: 8213). In some embodiments, the HC CDR2 has an amino acid sequence YINPYTGDIKYNERFRG (SEQ ID NO: 8217). In some embodiments, the HC CDR2 has an amino acid sequence FINPYNDDIQYNERFRG (SEQ ID NO.8221). In some embodiments, the HC CDR2 has an amino acid sequence TINPYNAEIKYNQKFQG (SEQ ID NO: 8222). In some embodiments, the HC CDR2 has an amino acid sequence TINPYNRDIQYNQKFQG (SEQ ID NO: 8225). In some embodiments, the HC CDR2 has an amino acid sequence FINPYNRDIKYNERFRG (SEQ ID NO: 8228). In some embodiments, the HC CDR2 has an amino acid sequence AINPYTNDIKYNERFRG (SEQ ID NO: 8229). In some embodiments, the HC CDR2 has an amino acid sequence AINPYTNHIQYNERFRG (SEQ ID NO: 8230). In some embodiments, the HC CDR2 has an amino acid sequence AINPYTRAIKYNERFRG (SEQ ID NO: 8231). In some embodiments, the HC CDR2 has an amino acid sequence TINPYNGDIQYNERFRG (SEQ ID NO: 8232). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNTDIKYNERFRG (SEQ ID NO: 8233). In some embodiments, the HC CDR2 has an amino acid sequence YINPYNGAIKYNQKFQG (SEQ ID NO: 8234). In some embodiments, the HC CDR2 has an amino acid sequenceWSGR Docket No.53676-767.601 AINPYNDDIQSNERFRG (SEQ ID NO: 8235). In some embodiments, the HC CDR2 has an amino acid sequence FINPYNRAIQYNQKFQG (SEQ ID NO: 8236). In some embodiments, the HC CDR2 has an amino acid sequence FINPYTNEIQYNERFRG (SEQ ID NO: 8237). In some embodiments, the HC CDR2 has an amino acid sequence YINPYNHDIQYNQKFQG (SEQ ID NO: 8237). In some embodiments, the HC CDR2 has an amino acid sequence SINPYTHDIQYNERFRG (SEQ ID NO: 8238). In some embodiments, the HC CDR2 has an amino acid sequence YINPYKNAIQYNQKFQG (SEQ ID NO: 8239). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNTDIQYNERFRG (SEQ ID NO: 8240). In some embodiments, the HC CDR2 has an amino acid sequence SINPYNGDIQYNERFRG (SEQ ID NO: 8241). In some embodiments, the HC CDR2 has an amino acid sequence TINPYNHDAQYNERFRG (SEQ ID NO: 8242). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNDDIKYNERFRG (SEQ ID NO: 8243). In some embodiments, the HC CDR2 has an amino acid sequence YINPYTHEIKYNERFRG (SEQ ID NO: 8245). In some embodiments, the HC CDR2 has an amino acid sequence FINPYKDDIKYNERFRG (SEQ ID NO: 8246). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNDDIKYNQKFQG (SEQ ID NO: 8247). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNRDIKYNERFRG (SEQ ID NO: 8248). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNGDIKYNERFRG (SEQ ID NO: 8249). In some embodiments, the HC CDR2 has an amino acid sequence YINPYTRDIKYNERFRG (SEQ ID NO: 8250). In some embodiments, the HC CDR2 has an amino acid sequence TINPYNTDIKYNERFRG (SEQ ID NO: 8251). In some embodiments, the HC CDR2 has an amino acid sequence TINPYNNDIQYNERFRG (SEQ ID NO: 8252). In some embodiments, the HC CDR2 has an amino acid sequence YINPYNGNIQYNERFRG (SEQ ID NO: 8253). In some embodiments, the HC CDR2 has an amino acid sequence AINPYTNEIQYNERFRG (SEQ ID NO: 8254). In some embodiments, the HC CDR2 has an amino acid sequence SINPYNHDIKYNERFRG (SEQ ID NO: 8255). In some embodiments, the HC CDR2 has an amino acid sequence FINPYKNEIKYNERFRG (SEQ ID NO: 8256). In some embodiments, the HC CDR2 has an amino acid sequence YINPYNNEIQYNERFRGR (SEQ ID NO: 8257) or a sequence SINPYNRHIQYNERFRG (SEQ ID NO: 8258) or a sequence SINPYTREIQYNERFRG (SEQ ID NO: 8259). In some embodiments, the HC CDR2 has an amino acid sequence FINPYTNDIQYNERFRG (SEQ ID NO: 8260). In some embodiments, the HC CDR2 has an amino acid sequence AINPYTNEIKYNERFRG (SEQ ID NO: 8261). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNDDIQYNERFRG (SEQ ID NO: 8263). In some embodiments, the HC CDR2 has an amino acid sequence YINPYNNDIKYNQKFQG (SEQ ID NO: 8264) or an amino acid sequence TINPYTREIQYNQKFQG (SEQ ID NO: 8266) or YINPYNNEIQYNQKFQG (SEQ ID NO: 8267). In some embodiments, the HC CDR2 has an amino acid sequence AINPYNHEIQYNQKFQG (SEQ ID NO: 8268). In some embodiments, the HC CDR2 has an amino acid sequence TINPYKHHIKYNERFRG (SEQ ID NO: 8269). In some embodiments, the HC CDR2 has an amino acid sequence FINPYTRAIKYNERFRG (SEQ ID NO: 8270). In someWSGR Docket No.53676-767.601 embodiments, the HC CDR2 has an amino acid sequence SINPYTRHIQYNERFRG (SEQ ID NO: 8273).

[0545] In one embodiment, the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GNGKWGDGAYRFFDF (SEQ ID NO: 7403). In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GEGKWGDGAYRFFDF (SEQ ID NO: 8046) In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GAGKWGDGAYRFFDF (SEQ ID NO: 8047). In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GNGKWGDGAYRFFDF (SEQ ID NO.7403). In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GNGKWGDGAYRFFDL (SEQ ID NO.8227). In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence LGNGKWGDGAYRFFDL (SEQ ID NO: 8224). In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence MGNGKWGDGAYRFFDL (SEQ ID NO: 8220). In some embodiments the heavy chain of a TRBC2 binding domain comprises a HC CDR3 having an amino acid sequence GNGKWGDGAYRFFDF (SEQ ID NO: 7403).

[0546] In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR1 having an amino acid sequence RSSENLVHSNGRTYLQ (SEQ ID NO: 8048) or RSSQNLVHSNGRTYLQ (SEQ ID NO: 7410). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR1 having an amino acid sequence RSSQNLVHSNARTYLQ (SEQ ID NO: 8276). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR2 having an amino acid sequence RVSNRFP (SEQ ID NO: 224).

[0547] In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSSLEPYT (SEQ ID NO: 8050) or SQSSYVPFT (SEQ ID NO: 8214). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSTYEPFT (SEQ ID NO: 8223). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR1 having an amino acid sequence RSSKNLVHSNGRTYLQ (SEQ ID NO: 8051). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR1 having an amino acid sequence RSSKNLVHSNARTYLQ (SEQ ID NO: 8271). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSTREPYT (SEQ ID NO: 8052) or SQSTHVPYT (SEQ ID NO: 225). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSTHLPYT (SEQ ID NO: 8262). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSTQEPYT (SEQ ID NO: 8265). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSSLLPYTF (SEQ ID NO: 8274). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSTLEPFT (SEQ ID NO: 8277). In one embodiment, the light chain of a TRBC2 binding domain comprises a LC CDR3 having an amino acid sequence SQSSHIPYT (SEQ IDWSG...

Claims

WSGR Docket No.53676-767.601 CLAIMS WHAT IS CLAIMED IS; 1. A composition comprising an agent that comprises an antigen binding domain that binds to NKp30, wherein the antigen binding domain comprises a heavy chain variable region (VH) comprising: (i) a heavy chain complementarity determining region 3 (VHCDR3) comprising the sequence of GDWHYFDY (SEQ ID NO: 504); and (ii) a heavy chain complementarity determining region 1 (VHCDR1) comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), a heavy chain complementarity determining region 2 (VHCDR2) comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), or a combination thereof.

2. The composition of claim 1, wherein the antigen binding domain comprises a VH comprising the VHCDR1 comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), the VHCDR2 comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), and the VHCDR3 comprising the sequence of GDWHYFDY (SEQ ID NO: 504).

3. The composition of claim 1 or 2, wherein the antigen binding domain comprises a light chain variable region (VL) comprising a light chain complementarity determining region 1 (VLCDR1) comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a light chain complementarity determining region 2 (VLCDR2) comprising the sequence of ENDRRPS (SEQ ID NO: 28), a light chain complementarity determining region 3 (VLCDR3) comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86), or any combination thereof.

4. The composition of any one of claims 1-3, wherein the antigen binding domain comprises a VL comprising a VLCDR1 comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a VLCDR2 comprising the sequence of ENDRRPS (SEQ ID NO: 28), and a VLCDR3 comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86).

5. The composition of any one of claims 1-4, wherein the antigen binding domain comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

505.

6. The composition of any one of claims 1-5, wherein the antigen binding domain comprises a VH comprising the sequence of SEQ ID NO:

505.

7. The composition of any one of claims 1-6, wherein the antigen binding domain comprises a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

13.

8. The composition of any one of claims 1-7, wherein the antigen binding domain comprises a VL comprising the sequence of SEQ ID NO:

13.

9. The composition of any one of claims 1-8, wherein the antigen binding domain comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 505 and a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

13.

10. The composition of any one of claims 1-9, wherein the antigen binding domain comprises a VH comprising the sequence of SEQ ID NO: 505 and a VL comprising the sequence of SEQ ID NO: 13.WSGR Docket No.53676-767.601 11. The composition of any one of claims 1-10, wherein the antigen binding domain is a full antibody or antigen-binding fragment thereof, a bivalent antibody, bispecific antibody, or a biparatopic antibody.

12. The composition of any one of claims 1-11, wherein the antigen binding domain is a Fab, F(ab')2, Fv, an single chain Fv (scFv), a diabody, a single domain antibody, or a camelid antibody.

13. The composition of any one of claims 1-12, wherein the antigen binding domain is a Fab or an scFv.

14. The composition of any one of claims 1-13, wherein the antigen binding domain comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or functional fragment thereof, IgG2 heavy chain constant region or functional fragment thereof, IgG3 heavy chain constant region or functional fragment thereof, IgGA1 heavy chain constant region or functional fragment thereof, IgGA2 heavy chain constant region or functional fragment thereof, IgG4 heavy chain constant region or functional fragment thereof, IgJ heavy chain constant region or functional fragment thereof, IgM heavy chain constant region or functional fragment thereof, IgD heavy chain constant region or functional fragment thereof, and IgE heavy chain constant region or functional fragment thereof.

15. The composition of claim 14, wherein the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to any one heavy chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

16. The composition of claim 14 or 15, wherein the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, or the sequence of SEQ ID NO:

511.

17. The composition of any one of claims 1-16, wherein the antigen binding domain comprises a kappa light chain constant region or functional fragment thereof, a lambda light chain constant region or functional fragment thereof, or a combination thereof.

18. The composition of claim 17, wherein the kappa light chain constant region comprises a sequence having at least 75% sequence identity to any one light chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

19. The composition of claim 17 or 18, wherein the kappa light chain constant region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 39 or the sequence of SEQ ID NO: 3644.

20. The composition of any one of claims 1-19, wherein the agent is a polypeptide molecule.

21. The composition of any one of claims 1-20, wherein the agent is a multifunctional molecule.WSGR Docket No.53676-767.601 22. The composition of any one of claims 1-21, wherein the agent is a multispecific molecule.

23. The composition of any one of claims 21 or 22, wherein the agent comprises at least one cytokine molecule or a functional fragment or variant thereof or a functional fragment or variant thereof, an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), a second antigen binding domain that binds to NKp30, an immune cell engager, a tumor-targeting moiety, or any combination thereof.

24. The composition of any one of claims 1-23, wherein the agent comprises at least two non-contiguous polypeptide chains comprising a first polypeptide chain and a second polypeptide chain; wherein the first polypeptide chain comprises, from N-terminus to C-terminus, the following configuration: A-[a first member of a dimerization module]-C, And the second polypeptide chain comprises, from N-terminus to C-terminus, the following configuration: B-[a second member of a dimerization module]-D; wherein the first member of the dimerization module comprises a first immunoglobulin constant domain; and the second member of the dimerization module comprises a second immunoglobulin constant domain; wherein at least one of A, B, C, and D is the antigen binding domain; and wherein the first polypeptide chain and the second polypeptide chain dimerize via interaction between the first member of the dimerization module and the second member of the dimerization module.

25. The composition of claim 24, wherein: (i) A is the antigen binding domain, and B, C, and D are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor-targeting moiety; (ii) B is the antigen binding domain, and A, C, and D are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor- targeting moiety; (iii) C is the antigen binding domain, and A, B, and D are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor- targeting moiety;WSGR Docket No.53676-767.601 (iv) D is the antigen binding domain, and A, B, and C are independently (a) absent, (b) at least one cytokine molecule or a functional fragment or variant thereof, (c) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC), (d) a second antigen binding domain that binds to NKp30, (e) an immune cell engager, or (f) a tumor- targeting moiety.

26. The composition of any one of claims 21-25, wherein the agent comprises at least one cytokine molecule or a functional fragment or variant thereof.

27. The composition of any one of claims 23-26, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof, IL- 7 or a functional fragment or variant thereof, IL-12 or a functional fragment or variant thereof, IL-15 or a functional fragment or variant thereof, IL-18 or a functional fragment or variant thereof, IL-21 or a functional fragment or variant thereof, interferon gamma or a functional fragment or variant thereof, or any combination thereof.

28. The composition of any one of claims 23-27, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

29. The composition of any one of claims 23-28, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

30. The composition of any one of claims 23-29, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof.

31. The composition of any one of claims 23-30, wherein the IL-2 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

32. The composition of any one of claims 23-31, wherein the IL-2 or a functional fragment or variant thereof comprises a sequence the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

33. The composition of any one of claims 23-29, wherein the at least one cytokine molecule or a functional fragment or variant thereof IL-15 or a functional fragment or variant thereof.

34. The composition of any one of claims 23-29 and 33, wherein the IL-15 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2170, SEQ ID NO: 3670, or SEQ ID NO: 3673.

35. The composition of any one of claims 23-29, 33, and 34, wherein the IL-15 or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2170, the sequence of SEQ ID NO: 3670, or the sequence of SEQ ID NO: 3673.WSGR Docket No.53676-767.601 36. The composition of any one of claims 23-29 and 33-35, wherein the at least one cytokine molecule or a functional fragment or variant thereof further comprises an IL15Ralpha dimerizing domain covalently linked the IL-15 or a functional fragment or variant thereof.

37. The composition of claim 36, wherein the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or a functional fragment or variant thereof.

38. The composition of claim 36 or 37, wherein the IL15Ralpha dimerizing domain comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3472.

39. The composition of any one of claims 36-38, wherein the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

40. The composition of any one of claims 36-39, wherein the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

41. The composition of any one of claims 36-40, wherein the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

42. The composition of any one of claims 23-29 and 33-41, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3474, SEQ ID NO: 3671, or SEQ ID NO: 3674.

43. The composition of any one of claims 23-29 and 33-42, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3474, the sequence of SEQ ID NO: 3671, or the sequence of SEQ ID NO: 3674.

44. The composition of any one of claims 23-29, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-7 or a functional fragment or variant thereof.

45. The composition of any one of claims 23-29 and 44, wherein the IL-7 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

46. The composition of any one of claims 23-29, 44, and 45, wherein the IL-7 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

47. The composition of any one of claims 23-29, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-12 or a functional fragment or variant or variant thereof.

48. The composition of any one of claims 23-29 and 47, wherein the IL-12 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

49. The composition of any one of claims 23-29, 47, and 48, wherein the IL-12 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

50. The composition of any one of claims 23-29, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-18 of a functional fragment or variant thereof.WSGR Docket No.53676-767.601 51. The composition of any one of claims 23-29 and 50, wherein the IL-18 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2192.

52. The composition of any one of claims 23-29, 50, and 51, wherein the IL-18 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2192.

53. The composition of any one of claims 23-29, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-21 or a functional fragment or variant thereof.

54. The composition of any one of claims 23-29 and 53, wherein the IL-21 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

55. The composition of any one of claims 23-29, 53, and 54, wherein the IL-21 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

56. The composition of any one of claims 21-55, wherein the agent comprises an antigen binding domain that binds to TRBC.

57. The composition of any one of claims 23-56, wherein the antigen binding domain that binds to TRBC binds to TRBC1 or TRBC2.

58. The composition of claim 57, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

59. The composition of claim 57 or 58, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 2A, 2B, 3A, 3B, 5A, 5B, 7, 8, 16, 39, and 40; or (iv) any combination thereof.WSGR Docket No.53676-767.601 60. The composition of any one of claims 57-59, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 7, 8, 16, 39, and 40.

61. The composition of any one of claims 57-60, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 7, 8, 16, 39, and 40.

62. The composition of any one of claims 57-60, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

63. The composition of any one of claims 57-62, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 4, 6, 7, 8, 16, 39, and 40; or (iv) any combination thereof.

64. The composition of any one of claims 57-59, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 7, 8, 16, 39, and 40.

65. The composition of any one of claims 57-60, wherein the antigen binding domain that binds to TRBC binds to TRBC1, and wherein the antigen binding domain that binds to TRBC comprises a VLWSGR Docket No.53676-767.601 comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 7, 8, 16, 39, and 40.

66. The composition of claim 57, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 9A, 9B, 11, 12, 13, 14, 15, 17, 39, and 40; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 9A, 9B, 12, 14, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

67. The composition of claim 57 or 66, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 9A, 9B, 12, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

68. The composition of any one of claims 57, 66, and 67, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 11, 15, 17, 39, and 40.

69. The composition of any one of claims 57 and 66-68, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 11, 15, 17, 39, and 40.

70. The composition of any one of claims 57 and 66-69, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40;WSGR Docket No.53676-767.601 (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

71. The composition of any one of claims 57 and 66-70, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 10, 12, 13, 14, 15, 17, 39, and 40; or (iv) any combination thereof.

72. The composition of any one of claims 57 and 66-71, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 11, 15, 17, 39, and 40.

73. The composition of any one of claims 57 and 66-72, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 11, 15, 17, 39, and 40.

74. The composition of any one of claims 57 and 66-73, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the scFv sequences listed in Tables 12, 15, 17, 39, and 40.

75. The composition of any one of claims 57 and 66-74, wherein the antigen binding domain that binds to TRBC binds to TRBC2, and wherein the antigen binding domain that binds to TRBC comprises a VL comprising any one sequence selected from the group consisting of the scFv sequences listed in Tables 12, 15, 17, 39, and 40.

76. The composition of any one of claims 21-75, wherein the agent comprises a second antigen binding domain that binds to NKp30.

77. The composition of any one of claims 23-76, wherein the antigen binding domain and the second antigen binding domain that binds to NKp30 are same.WSGR Docket No.53676-767.601 78. The composition of any one of claims 23-76, wherein the antigen binding domain and the second antigen binding domain that binds to NKp30 are different.

79. The composition of any one of claims 23-77, wherein the second antigen binding domain that binds to NKp30 comprises a heavy chain variable region (VH) comprising: (i) a heavy chain complementarity determining region 3 (VHCDR3) comprising the sequence of GDWHYFDY (SEQ ID NO: 504); and (ii) a heavy chain complementarity determining region 1 (VHCDR1) comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), a heavy chain complementarity determining region 2 (VHCDR2) comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), or a combination thereof.

80. The composition of any one of claims 23-77 and 79, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising the VHCDR1 comprising the sequence of GFSIIRVGYHWN (SEQ ID NO: 502), the VHCDR2 comprising the sequence of YIHRSGSTSYNPSLKS (SEQ ID NO: 503), and the VHCDR3 comprising the sequence of GDWHYFDY (SEQ ID NO: 504).

81. The composition of any one of claims 23-77, 79, and 80, wherein the second antigen binding domain that binds to NKp30 comprises a light chain variable region (VL) comprising a light chain complementarity determining region 1 (VLCDR1) comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a light chain complementarity determining region 2 (VLCDR2) comprising the sequence of ENDRRPS (SEQ ID NO: 28), a light chain complementarity determining region 3 (VLCDR3) comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86), or any combination thereof.

82. The composition of any one of claims 23-77 and 79-81, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising a VLCDR1 comprising the sequence of SGEKLSDKYVH (SEQ ID NO: 26), a VLCDR2 comprising the sequence of ENDRRPS (SEQ ID NO: 28), and a VLCDR3 comprising the sequences of QFWDSTNSAV (SEQ ID NO: 86).

83. The composition of any one of claims 23-77 and 79-82, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

505.

84. The composition of any one of claims 23-77 and 79-83, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising the sequence of SEQ ID NO:

505.

85. The composition of any one of claims 23-77 and 79-84, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

13.

86. The composition of any one of claims 23-77 and 79-85, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising the sequence of SEQ ID NO:

13.

87. The composition of any one of claims 23-77 and 79-86, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising a sequence having at least 75% sequence identity toWSGR Docket No.53676-767.601 the sequence of SEQ ID NO: 505 and a VL comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

13.

88. The composition of any one of claims 23-77 and 79-87, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising the sequence of SEQ ID NO: 505 and a VL comprising the sequence of SEQ ID NO:

13.

89. The composition of any one of claims 23-76 and 78, wherein the second antigen binding domain that binds to NKp30 comprises a heavy chain variable region (VH) comprising: (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; or (iv) any combination thereof.

90. The composition of any one of claims 23-76, 78, and 89, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 20A, 20B, 21A, 21B, 23A, 23B, 25, 26, 16, and 17; or (iv) any combination thereof.

91. The composition of any one of claims 23-76, 78, 89, and 90, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 25, 26, 16, and 17.

92. The composition of any one of claims 23-76, 78, and 89-91, wherein the second antigen binding domain that binds to NKp30 comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 25, 26, 16, and 17.

93. The composition of any one of claims 23-76, 78, and 89-92, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 22, 24, 25, 26, 16, and 17;WSGR Docket No.53676-767.601 (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 22, 24, 25, 26, 16, and 17; or (iv) any combination thereof.

94. The composition of any one of claims 23-76, 78, and 89-93, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 22, 24, 25, 26, 16, and 17; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 22, 24, 25, 26, 16, and 17; or (iv) any combination thereof.

95. The composition of any one of claims 23-76, 78, and 89-94, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 25, 26, 16, and 17.

96. The composition of any one of claims 23-76, 78, and 89-95, wherein the second antigen binding domain that binds to NKp30 comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 25, 26, 16, and 17.

97. The composition of any one of claims 21-96, wherein the agent comprises an immune cell engager.

98. The composition of any one of claims 23-97, wherein the immune cell engager is selected from the group consisting of a T cell engager, an NK cell engager, a B cell engager, a dendritic cell engager, and the macrophage cell engager.

99. The composition of any one of claims 21-98, wherein the agent comprises a T cell engager.

100. The composition of claim 98 or 99, wherein the T cell engager comprises an antigen binding domain that binds to a T cell receptor beta chain region (TCRβV).

101. The composition of claim 100, wherein the antigen binding domain that binds to a T cell receptor beta chain region (TCRβV) binds to a TCRβV of any one T cell receptor beta chain (TCRβ) subfamily member selected from the group consisting of TCRβ subfamily members listed in Tables 29 and 42A.

102. The composition of claim 100 or 101, wherein the antigen binding domain that binds to a TCRβV comprises a heavy chain variable region (VH) comprising:WSGR Docket No.53676-767.601 (i) a VHCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VHCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VHCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

103. The composition of any one of claims 100-102, wherein the antigen binding domain that binds to a TCRβV comprises a VH comprising: (i) a VHCDR1 comprising to any one sequence selected from the group consisting of the VHCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VHCDR2 comprising any one sequence selected from the group consisting of the VHCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VHCDR3 comprising any one sequence selected from the group consisting of the VHCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

104. The composition of any one of claims 100-103, wherein the antigen binding domain that binds to a TCRβV comprises a VH comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VH sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

105. The composition of any one of claims 100-104, wherein the antigen binding domain that binds to a TCRβV comprises a VH comprising any one sequence selected from the group consisting of the VH sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

106. The composition of any one of claims 100-105, wherein the antigen binding domain that binds to a TCRβV comprises a VL comprising: (i) a VLCDR1 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VLCDR2 comprising a sequence having at least 75% sequence identity any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VLCDR3 comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.WSGR Docket No.53676-767.601 107. The composition of any one of claims 100-106, wherein the antigen binding domain that binds to a TCRβV comprises a VL comprising: (i) a VLCDR1 comprising any one sequence selected from the group consisting of the VLCDR1 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (ii) a VLCDR2 comprising any one sequence selected from the group consisting of the VLCDR2 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; (iii) a VLCDR3 comprising any one sequence selected from the group consisting of the VLCDR3 sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51; or (iv) any combination thereof.

108. The composition of any one of claims 100-107, wherein the antigen binding domain that binds to a TCRβV comprises a VL comprising a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the VL sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

109. The composition of any one of claims 100-108, wherein the antigen binding domain that binds to a TCRβV comprises a VL comprising any one sequence selected from the group consisting of the VL sequences listed in Tables 30, 32, 42B, 42C, 43A, 43B, and 43-51.

110. The composition of any one of claims 21-98, wherein the agent comprises a NK cell engager.

111. The composition of claim 98 or 110, wherein the NK cell engager does not bind to NKp30.

112. The composition of any one of claims 98, 110, and 111, wherein the NK cell engager comprise a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequences listed in Table 27.

113. The composition of any one of claims 98 and 110-112, wherein the NK cell engager comprise any one sequence selected from the group consisting of the sequences listed in Table 27.

114. The composition of any one of claims 21-113, wherein the agent comprises a tumor-targeting moiety.

115. The composition of any one of claims 23-114, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor- targeting moiety, or any combination thereof is a full antibody or antigen-binding fragment thereof, a bivalent antibody, bispecific antibody, or a biparatopic antibody.

116. The composition of any one of claims 23-115, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor- targeting moiety, or any combination thereof is a Fab, F(ab')2, Fv, an single chain Fv (scFv), a diabody, a single domain antibody, or a camelid antibody.

117. The composition of any one of claims 23-116, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor- targeting moiety, or any combination thereof is a Fab or an scFv.

118. The composition of any one of claims 23-117, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor-WSGR Docket No.53676-767.601 targeting moiety, or any combination thereof comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or functional fragment thereof, IgG2 heavy chain constant region or functional fragment thereof, IgG3 heavy chain constant region or functional fragment thereof, IgGA1 heavy chain constant region or functional fragment thereof, IgGA2 heavy chain constant region or functional fragment thereof, IgG4 heavy chain constant region or functional fragment thereof, IgJ heavy chain constant region or functional fragment thereof, IgM heavy chain constant region or functional fragment thereof, IgD heavy chain constant region or functional fragment thereof, and IgE heavy chain constant region or functional fragment thereof.

119. The composition of claim 118, wherein the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to any one heavy chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

120. The composition of claim 118 or 119, wherein the one or more heavy chain constant regions comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, or the sequence of SEQ ID NO:

511.

121. The composition of any one of claims 118-120, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, the tumor- targeting moiety, or any combination thereof comprises a kappa light chain constant region or functional fragment thereof, a lambda light chain constant region or functional fragment thereof, or a combination thereof.

122. The composition of claim 121, wherein the kappa light chain constant region comprises a sequence having at least 75% sequence identity to any one light chain constant region sequences listed in Tables 3, 7, 8, 16, 17, 39, 40, 42C, 43A, 43B, and 46-51.

123. The composition of claim 121 or 122, wherein the kappa light chain constant region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 39 or the sequence of SEQ ID NO: 3644.

124. The composition of any one of claims 24-123, wherein the first member of the dimerization module comprises a first Fc region and the second member of the dimerization module comprises a second Fc region.

125. The composition of claim 124, wherein the first Fc region or the second Fc region comprises N297.WSGR Docket No.53676-767.601 126. The composition of claim 124, wherein the first Fc region is selected from the group consisting of an IgG1 Fc region or a functional fragment thereof, an IgG2 Fc region or a functional fragment thereof, an IgG3 Fc region or a functional fragment thereof, an IgGA1 Fc region or a functional fragment thereof, an IgGA2 Fc region or a functional fragment thereof, an IgG4 Fc region or a functional fragment thereof, an IgJ Fc region or a functional fragment thereof, an IgM Fc region or a functional fragment thereof, an IgD Fc region or a functional fragment thereof, and an IgE Fc region or a functional fragment thereof.

127. The composition of claim 124 or 126, wherein the second Fc region is selected from the group consisting of an IgG1 Fc region or a functional fragment thereof, an IgG2 Fc region or a functional fragment thereof, an IgG3 Fc region or a functional fragment thereof, an IgGA1 Fc region or a functional fragment thereof, an IgGA2 Fc region or a functional fragment thereof, an IgG4 Fc region or a functional fragment thereof, an IgJ Fc region or a functional fragment thereof, an IgM Fc region or a functional fragment thereof, an IgD Fc region or a functional fragment thereof, and an IgE Fc region or a functional fragment thereof.

128. The composition of any one of claims 124-127, wherein the first Fc region, the second Fc region, or a combination thereof comprises an Fc interface with one or more of: a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, wherein the dimerization of the first Fc region and the second Fc region is enhanced as indicated by a greater ratio of heteromultimer:homomultimer forms relative to a dimerization of Fc regions with a non-engineered interface.

129. The composition of claim 128, wherein: (i) the first Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering (b) a Cys at position 349 in a heavy chain constant region according to EU Numbering), (c) a Ser at position 366 in a heavy chain constant region according to EU Numbering), (d) an Ala at position 368 in a heavy chain constant region according to EU Numbering); and (e) a Val at position 407 in a heavy chain constant region according to EU Numbering); and (ii) the second Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering), (b) a Cys at position 354 in a heavy chain constant region according to EU Numbering), and (c) a Trp at position 366 in a heavy chain constant region according to EU Numbering).

130. The composition of claim 128, wherein: (i) the first Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering), (b) a Cys at position 354 in a heavy chain constant region according to EU Numbering), andWSGR Docket No.53676-767.601 (c) a Trp at position 366 in a heavy chain constant region according to EU Numbering); and (ii) the second Fc region comprises: (a) an Ala at position 297 in a heavy chain constant region according to EU Numbering (b) a Cys at position 349 in a heavy chain constant region according to EU Numbering), (c) a Ser at position 366 in a heavy chain constant region according to EU Numbering), (d) an Ala at position 368 in a heavy chain constant region according to EU Numbering); and (e) a Val at position 407 in a heavy chain constant region according to EU Numbering).

131. The composition of any one of claims 124-130, wherein the first Fc region comprises a mutation that decreases Fc receptor binding to the agent relative to an agent without the mutation.

132. The composition of any one of claims 124-131, wherein the first Fc region comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation, or a combination thereof according to EU Numbering.

133. The composition of any one of claims 124-132, wherein the second Fc region comprises a mutation that decreases Fc receptor binding to the agent relative to an agent without the mutation.

134. The composition of any one of claims 124-133, wherein the second Fc region comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation, or a combination thereof according to EU Numbering.

135. The composition of any one of claims 124-134, wherein the first Fc region comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 4663, the sequence of SEQ ID NO: 4664, the sequence of SEQ ID NO: 4665, the sequence of SEQ ID NO: 4666, the sequence of SEQ ID NO: 4667, the sequence of SEQ ID NO: 4668, the sequence of SEQ ID NO: 4669, and the sequence of SEQ ID NO: 4670.

136. The composition of any one of claims 124-135, wherein the second Fc region comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO:WSGR Docket No.53676-767.601 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 4663, the sequence of SEQ ID NO: 4664, the sequence of SEQ ID NO: 4665, the sequence of SEQ ID NO: 4666, the sequence of SEQ ID NO: 4667, the sequence of SEQ ID NO: 4668, the sequence of SEQ ID NO: 4669, and the sequence of SEQ ID NO: 4670.

137. The composition of any one of claims 124-136, wherein the first Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 4663, the sequence of SEQ ID NO: 4664, the sequence of SEQ ID NO: 4665, the sequence of SEQ ID NO: 4666, the sequence of SEQ ID NO: 4667, the sequence of SEQ ID NO: 4668, the sequence of SEQ ID NO: 4669, and the sequence of SEQ ID NO: 4670.

138. The composition of any one of claims 124-137, wherein the second Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 73, the sequence of SEQ ID NO: 76, the sequence of SEQ ID NO: 74, the sequence of SEQ ID NO: 75, the sequence of SEQ ID NO: 40, the sequence of SEQ ID NO: 41, the sequence of SEQ ID NO: 42, the sequence of SEQ ID NO: 3645, the sequence of SEQ ID NO: 3646, the sequence of SEQ ID NO: 3649, the sequence of SEQ ID NO: 3451, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3647, the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 513, the sequence of SEQ ID NO: 514, the sequence of SEQ ID NO: 515, the sequence of SEQ ID NO: 516, the sequence of SEQ ID NO: 3447, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 4663, the sequence of SEQ ID NO: 4664, the sequence of SEQ ID NO: 4665, the sequence of SEQ ID NO: 4666, the sequence of SEQ ID NO: 4667, the sequence of SEQ ID NO: 4668, the sequence of SEQ ID NO: 4669, and the sequence of SEQ ID NO: 4670.

139. The composition of any one of claims 124-136, wherein the first Fc region comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3477, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 515, and the sequence of SEQ ID NO: 516.WSGR Docket No.53676-767.601 140. The composition of any one of claims 124-136 and 139, wherein the second Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648, the sequence of SEQ ID NO: 508, the sequence of SEQ ID NO: 506, the sequence of SEQ ID NO: 507, the sequence of SEQ ID NO: 3477, the sequence of SEQ ID NO: 511, the sequence of SEQ ID NO: 515, or the sequence of SEQ ID NO:

516.

141. The composition of any one of claims 124-136, 139, and 140, wherein the first Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 3648, SEQ ID NO: 508, SEQ ID NO: 506, SEQ ID NO: 507, SEQ ID NO: 3477, SEQ ID NO: 511, SEQ ID NO: 515, or SEQ ID NO:

516.

142. The composition of any one of claims 124-136 and 139-141, wherein the second Fc region comprises any one sequence selected from the group consisting of the sequence of SEQ ID NO: 3648, SEQ ID NO: 508, SEQ ID NO: 506, SEQ ID NO: 507, SEQ ID NO: 3477, SEQ ID NO: 511, SEQ ID NO: 515, or SEQ ID NO:

516.

143. The composition of any one of claims 124-142, wherein: (i) the first Fc region comprises the sequence of SEQ ID NO: 506 or the sequence of SEQ ID NO: 507, and the second Fc region comprises the sequence of SEQ ID NO: 3648 or the sequence of SEQ ID NO: 508; or (ii) the first Fc region comprises the sequence of SEQ ID NO: 3447 or the sequence of SEQ ID NO: 511, and the second Fc region comprises the sequence of SEQ ID NO: 515 or the sequence of SEQ ID NO:

516.

144. The composition of any one of claims 124-142, wherein: (i) the first Fc region comprises the sequence of SEQ ID NO: 3648 or the sequence of SEQ ID NO: 508, and the second Fc region comprises the sequence of SEQ ID NO: 506 or the sequence of SEQ ID NO: 507; or (ii) the first Fc region comprises the sequence of SEQ ID NO: 515 or the sequence of SEQ ID NO: 516, and the second Fc region comprises the sequence of SEQ ID NO: 3447 or the sequence of SEQ ID NO:

511.

145. The composition of any one of claims 1-144, wherein the agent comprises a first polypeptide, a second polypeptide, and at least one cytokine molecule or a functional fragment or variant thereof, wherein the first polypeptide and the second polypeptide are non-contiguous; wherein: (i) the first polypeptide comprises a first member of the dimerization module linked to: (a) the antigen binding domain comprising a VH and a VL, or (b) a first portion of the antigen binding domain comprising the VH, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising a VL, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module, andWSGR Docket No.53676-767.601 wherein the at least one cytokine molecule or a functional fragment or variant thereof is covalently linked to the second polypeptide or the first polypeptide.

146. The composition of any one of claims 1-145, wherein the agent comprises a first polypeptide, a second polypeptide, and at least one cytokine molecule or a functional fragment or variant thereof, wherein the first polypeptide and the second polypeptide are non-contiguous; wherein: (i) the first polypeptide comprises a first member of a dimerization module linked to a first portion of the antigen binding domain comprising the VH, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising the VL, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module, and wherein the at least one cytokine molecule or a functional fragment or variant thereof is covalently linked to the second polypeptide.

147. The composition of claim 146, wherein the at least one cytokine molecule or a functional fragment or variant thereof is covalently linked to the N-terminus of the second polypeptide, the C-terminus of the second polypeptide, or a combination thereof.

148. The composition of claim 146 or 147, wherein the at least one cytokine molecule or a functional fragment or variant thereof is within a single contiguous polypeptide chain of the second polypeptide.

149. The composition of any one of claims 145-148, wherein the first polypeptide comprises: (i) the antigen binding domain comprising the VH and the VL, wherein the antigen binding domain further comprises a heavy chain constant domain 1 (CH1) linked to the VH; or (ii) the first portion of the first antigen binding domain comprising the VH, wherein the first portion of the antigen binding domain further comprises a CH1 linked to the VH.

150. The composition of claim 149, wherein the CH1 is linked to the C-terminus of the VH.

151. The composition of any one of claims 145-150, wherein the agent comprises: (i) the first polypeptide comprising the antigen binding domain that comprises the VH and the VL, wherein the antigen binding domain further comprises a light chain constant domain (CL) linked to the VL; or (ii) the first polypeptide comprising the first portion of the antigen binding domain and the third polypeptide comprising the second portion of the antigen binding domain, wherein the second portion of the antigen binding domain further comprises a CL linked to the VL.

152. The composition of claim 151, wherein the CL is linked to the C-terminus of the VL.

153. The composition of any one of claims 145-152, wherein the first member of the dimerization module is linked to the C-terminus of the antigen binding domain comprising the VH and the VL, or the C-terminus of the first portion of the first antigen binding domain comprising the VH.

154. The composition of any one of claims 145-153, wherein the second polypeptide comprises:WSGR Docket No.53676-767.601 (i) an antigen binding domain that binds TRBC, an second antigen binding domain that binds to NKp30, an immune cell engager, or an tumor-targeting moiety comprising a VH and a VL, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a heavy chain constant domain 1 (CH1) linked to the VH; or (ii) the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising the VH, wherein the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a CH1 linked to the VH.

155. The composition of claim 154, wherein the CH1 is linked to the C-terminus of the VH.

156. The composition of any one of claims 145-155, wherein the agent comprises: (i) the second polypeptide comprising an antigen binding domain that binds TRBC, an second antigen binding domain that binds to NKp30, an immune cell engager, or an tumor-targeting moiety that comprises a VH and a VL, wherein the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety further comprises a light chain constant domain (CL) linked to the VL; or (ii) the second polypeptide comprising the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety, wherein the agent further comprises a fourth polypeptide comprising the second portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety, wherein the fourth polypeptide is non-contiguous with the first polypeptide, the second polypeptide, and third polypeptide, and wherein the second portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor- targeting moiety further comprises a CL linked to the VL.

157. The composition of claim 156, wherein the CL is linked to the C-terminus of the VL.

158. The composition of any one of claims 154-157, wherein the second member of the dimerization module is linked to the C-terminus of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising the VH and the VL, or the C-terminus of the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising the VH.

159. The composition of any one of claims 145-158, wherein the agent further comprises:WSGR Docket No.53676-767.601 (i) a linker between the first member of the dimerization module and the antigen binding domain comprising a VH and a VL, or the first portion of the antigen binding domain comprising a VH; (ii) a linker between the second member of the dimerization module and an antigen binding domain that binds TRBC, an second antigen binding domain that binds to NKp30, an immune cell engager, or an tumor-targeting moiety comprising a VH and a VL, or the first portion of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor-targeting moiety comprising a VH; (iii) a linker between the VH and the VL of the antigen binding domain; (iv) a linker between the CH1 and the VH of the antigen binding domain; (v) a linker between the CL and the VL of the antigen binding domain; (vi) a linker between the VH and the VL of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor- targeting moiety; (vii) a linker between the CH1 and the VH of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor- targeting moiety; (viii) a linker between the CL and the VL of the antigen binding domain that binds TRBC, the second antigen binding domain that binds to NKp30, the immune cell engager, or the tumor- targeting moiety; (ix) a linker between the at least one cytokine molecule or a functional fragment or variant thereof and the first polypeptide, a linker between the at least one cytokine molecule or a functional fragment or variant thereof and the second polypeptide, or any combination thereof; or (x) any combination thereof.

160. The composition of claim 159, wherein the linker is selected from the group consisting of a cleavable linker, a non-cleavable linker, a peptide linker, a flexible linker, a rigid linker, a helical linker, and a non-helical linker.

161. The composition of claim 160, wherein the linker is the peptide linker comprises the sequence of SEQ ID NO: 3473, SEQ ID NO: 3308, or SEQ ID NO: 8291.

162. The composition of any one of claims 145-161, wherein the at least one cytokine molecule or a functional fragment or variant thereof is linked to the N-terminus of the first polypeptide, the C- terminus of the first polypeptide, the N-terminus of the second polypeptide, the C-terminus of the second polypeptide, the N-terminus of the third polypeptide, the C-terminus of the third polypeptide, or any combination thereof.

163. The composition of claim 162, wherein the at least one cytokine molecule or a functional fragment or variant thereof is within a single contiguous polypeptide chain of the first polypeptide, the secondWSGR Docket No.53676-767.601 polypeptide, or the third cytokine polypeptide to which the at least one cytokine molecule or a functional fragment or variant thereof is linked.

164. The composition of any one of claims 145-163, wherein the agent comprises: (i) the first polypeptide comprising the first portion of the dimerization module linked to the C- terminus of the first portion of the antigen binding domain; (ii) the second polypeptide comprising the second portion of the dimerization module; (iii) the third polypeptide comprising the second portion of the antigen binding domain; and (iv) the at least one cytokine molecule or a functional fragment or variant thereof covalently linked to the N-terminus of the second polypeptide.

165. The composition of any one of claims 24-164, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3472, the sequence of SEQ ID NO: 2170, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

166. The composition of any one of claims 24-165, wherein the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 2170 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3644.

167. The composition of claim 166, wherein the second polypeptide chain comprises, from the N- terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 3473 operatively linked to the sequence of SEQ ID NO: 2170 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO:

508.

168. The composition of any one of claims 24-167, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3474, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

169. The composition of any one of claims 24-168, wherein the agent comprises:WSGR Docket No.53676-767.601 (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3474 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3644.

170. The composition of claim 169, wherein the second polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3474 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO:

508.

171. The composition of any one of claims 24-170, wherein the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3420; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

501.

172. The composition of any one of claims 24-171, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3420; and (iii) a third polypeptide comprising the sequence of SEQ ID NO:

501.

173. The composition of any one of claims 24-164, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3472, the sequence of SEQ ID NO: 3670, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

174. The composition of any one of claims 24-164 and 173, wherein the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 3670 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3644.WSGR Docket No.53676-767.601 175. The composition of claim 174, wherein the second polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3472 operatively linked to the sequence of SEQ ID NO: 3473 operatively linked to the sequence of SEQ ID NO: 3670 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO:

508.

176. The composition of any one of claims 24-164 and 173-175, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3671, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

177. The composition of any one of claims 24-164 and 173-176, wherein the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3671 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; (iii) a third polypeptide comprising, from the N-terminus to the C-terminus, comprising the sequence of SEQ ID NO: 13, and the sequence of SEQ ID NO: 3644.

178. The composition of claim 177, wherein the second polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 3671 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO:

508.

179. The composition of any one of claims 24-164 and 173-178, wherein the wherein the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3669; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

501.

180. The composition of any one of claims 24-164 and 173-179, wherein the wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3669; and (iii) a third polypeptide comprising the sequence of SEQ ID NO:

501.

181. The composition of any one of claims 24-164, wherein the agent comprises:WSGR Docket No.53676-767.601 (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, and the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising the sequence of 2270, and the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

182. The composition of any one of claims 24-164 and 181, wherein the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3451; (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of 2270 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 508; and (iii) a third polypeptide comprising the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3644.

183. The composition of claim 182, wherein the second polypeptide comprises from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 2270 operatively linked to the sequence of SEQ ID NO: 3308 operatively linked to the sequence of SEQ ID NO: 3648 or SEQ ID NO:

508.

184. The composition of any one of claims 24-164 and 181-183, wherein the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3340; and (iii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

501.

185. The composition of any one of claims 24-164 and 181-184, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 500; (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 3340; and (iii) a third polypeptide comprising the sequence of SEQ ID NO:

501.

186. The composition of any one of claims 1-145, wherein the agent comprises a first polypeptide and a second polypeptide, wherein the first polypeptide and the second polypeptide are non-contiguous, and wherein: (i) the first polypeptide comprises a first member of the dimerization module linked to: (a) the antigen binding domain comprising a VH of the antigen binding domain and a VL of the antigen binding domain, or (b) a first portion of the antigen binding domain comprising the VH of the antigen binding domain, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising a VL of the antigen bindingWSGR Docket No.53676-767.601 domain, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module linked to: (a) an antigen binding domain that binds to T cell receptor beta chain constant domain (TRBC) comprising a VH and a VL, or (b) a first portion of the antigen binding domain that binds to TRBC comprising the VH of the antigen binding domain that binds to TRBC, wherein the agent further comprises a fourth polypeptide comprising a second portion of the antigen binding domain that binds to TRBC comprising a VL of the antigen binding domain that binds to TRBC, wherein the fourth polypeptide is non-contiguous with the first polypeptide, the second polypeptide, and the third polypeptide.

187. The composition of any one of claims 1-145 and 186, wherein the agent comprises a first polypeptide and a second polypeptide, wherein the first polypeptide and the second polypeptide are non-contiguous, wherein: (i) the first polypeptide comprises a first member of the dimerization module linked to a first portion of the antigen binding domain comprising the VH of the antigen binding domain, wherein the agent further comprises a third polypeptide comprising a second portion of the antigen binding domain comprising a VL of the antigen binding domain, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide; and (ii) the second polypeptide comprises a second member of the dimerization module linked to an antigen binding domain that binds to TRBC.

188. The composition of any one of claims 1-145, 186, and 187, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 505, the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3447 or SEQ ID NO: 511; (ii) a third polypeptide comprising the sequence of SEQ ID NO: 8308 and the sequence of SEQ ID NO: 3644; and (iii) a second polypeptide chain comprising the sequence of SEQ ID NO: 517 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514; or a second polypeptide chain comprising the sequence of SEQ ID NO: 512 and the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

189. The composition of any one of claims 1-145 and 186-188, wherein the agent comprises: (i) a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 13 operatively linked to the sequence of SEQ ID NO: 3447 or SEQ ID NO: 511; (ii) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8308 operatively linked to the sequence of SEQ ID NO: 3644; and (iii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 517 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514; or a second polypeptide chain comprising, from the N-terminus to the C-terminus, theWSGR Docket No.53676-767.601 sequence of SEQ ID NO: 512 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

190. The composition of claim 189, wherein the first polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 8291 operatively linked to the sequence of SEQ ID NO:

13.

191. The composition of any one of claims 1-145 and 186-190, wherein the agent comprises: (i) a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 509; (ii) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 7380; and (iii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 7382,the sequence of SEQ ID NO: 4677, the sequence of SEQ ID NO: 7379, or the sequence of SEQ ID NO: 4675.

192. The composition of any one of claims 1-145 and 186-191, wherein the agent comprises: (i) a first polypeptide chain comprising the sequence of SEQ ID NO: 509 or the sequence of SEQ ID NO: 4704; (ii) a third polypeptide comprising the sequence of SEQ ID NO: 7380; and (iii) a second polypeptide chain comprising the sequence of SEQ ID NO: 7382,the sequence of SEQ ID NO: 4677, the sequence of SEQ ID NO: 7379, or the sequence of SEQ ID NO: 4675.

193. The composition of any one of claims 1-145, wherein the agent comprises a first polypeptide chain comprising the sequence of SEQ ID NO: 505, the sequence of SEQ ID NO: 13 and the sequence of SEQ ID NO: 3447 or SEQ ID NO:

511.

194. The composition of any one of claims 1-145 and 193, wherein the agent comprises a first polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 13 operatively linked to sequence of SEQ ID NO: 3447 or SEQ ID NO:

511.

195. The composition of claim 194, wherein the first polypeptide comprises, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 505 operatively linked to the sequence of SEQ ID NO: 8291 operatively linked to the sequence of SEQ ID NO:

13.

196. The composition of any one of claims 1-145 and 193-195, wherein the agent comprises a first polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO:

509.

197. The composition of any one of claims 1-145 and 193-196, wherein the agent comprises a first polypeptide chain comprising the sequence of SEQ ID NO:

509.

198. The composition of any one of claims 1-145 and 193-197, wherein the agent comprises a first polypeptide chain comprising the sequence of SEQ ID NO:

509.

199. The composition of any one of claims 193-198, wherein the agent comprises:WSGR Docket No.53676-767.601 (i) a third polypeptide comprising the sequence of SEQ ID NO: 7419 and the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 7420 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514 or a second polypeptide chain comprising the sequence of SEQ ID NO: 7423 and the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

200. The composition of any one of claims 193-199, wherein the agent comprises: (i) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 7419 operatively linked to the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 7420 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514 or a second polypeptide chain comprising the sequence of SEQ ID NO: 7423 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

201. The composition of any one of claims 193-200, wherein the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 7439; and (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 4680, the sequence of SEQ ID NO: 7438, the sequence of SEQ ID NO: 4681, or the sequence of SEQ ID NO: 7440.

202. The composition of any one of claims 193-201, wherein the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 7439; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 4680, the sequence of SEQ ID NO: 7438, the sequence of SEQ ID NO: 4681, or the sequence of SEQ ID NO: 7440.

203. The composition of any one of claims 193-198, wherein the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8012 and the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8011 and the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

204. The composition of any one of claims 193-198 and 203, wherein the agent comprises: (i) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8012 operatively linked to the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8011 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

205. The composition of any one of claims 193-198, 203, and 204, wherein the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8002; andWSGR Docket No.53676-767.601 (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8001.

206. The composition of any one of claims 193-198 and 203-205, wherein the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8002; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8001.

207. The composition of any one of claims 193-198, wherein the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8014 and the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8013 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, a second polypeptide chain comprising the sequence of SEQ ID NO: 8299 and the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, or a second polypeptide chain comprising the sequence of SEQ ID NO: 8303 and the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

208. The composition of any one of claims 193-198 and 207, wherein the agent comprises: (i) a third polypeptide comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8014 operatively linked to the sequence of SEQ ID NO: 3644; and (ii) a second polypeptide chain comprising, from the N-terminus to the C-terminus, the sequence of SEQ ID NO: 8013 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, a second polypeptide chain comprising the sequence of SEQ ID NO: 8299 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO: 514, or a second polypeptide chain comprising the sequence of SEQ ID NO: 8303 operatively linked to the sequence of SEQ ID NO: 513 or SEQ ID NO:

514.

209. The composition of any one of claims 193-198, 207, and 208, wherein the agent comprises: (i) a third polypeptide comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8005; and (ii) a second polypeptide chain comprising a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 8004, the sequence of SEQ ID NO: 4685, the sequence of SEQ ID NO: 8007, the sequence of SEQ ID NO: 4686, the sequence of SEQ ID NO: 4687, or the sequence of SEQ ID NO: 8009.

210. The composition of any one of claims 193-198 and 207-209, wherein the agent comprises: (i) a third polypeptide comprising the sequence of SEQ ID NO: 8005; and (ii) a second polypeptide chain comprising the sequence of SEQ ID NO: 8004, the sequence of SEQ ID NO: 4685, the sequence of SEQ ID NO: 8007, the sequence of SEQ ID NO: 4686, the sequence of SEQ ID NO: 4687, or the sequence of SEQ ID NO: 8009.

211. The composition of any one of claims 186-210, wherein the agent further comprises at least one cytokine molecule or a functional fragment or variant thereof, wherein the at least one cytokine molecule or a functional fragment or variant thereof is linked to the first member of the dimerizationWSGR Docket No.53676-767.601 module, the second member of the dimerization module, the antigen binding domain, the antigen binding domain that binds to TRBC, or any combination thereof.

212. The composition of claim 211, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof, IL-7 or a functional fragment or variant thereof, IL-12 or a functional fragment or variant thereof, IL-15 or a functional fragment or variant thereof, IL-18 or a functional fragment or variant thereof, IL-21 or a functional fragment or variant thereof, interferon gamma or a functional fragment or variant thereof, or any combination thereof.

213. The composition of claim 211 or 212, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

214. The composition of any one of claims 211-213, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises any one sequence selected from the group consisting of the cytokine sequences listed in Tables 40, 42C, 43A, 43B, and 46-51.

215. The composition of any one of claims 211-214, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-2 or a functional fragment or variant thereof.

216. The composition of any one of claims 212-215, wherein the IL-2 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

217. The composition of any one of claims 212-216, wherein the IL-2 or a functional fragment or variant thereof comprises a sequence the sequence of SEQ ID NO: 2191, the sequence of SEQ ID NO: 2270, or the sequence of SEQ ID NO: 2280.

218. The composition of any one of claims 211-214, wherein the at least one cytokine molecule or a functional fragment or variant thereof IL-15 or a functional fragment or variant thereof.

219. The composition of any one of claims 212-214 and 218, wherein the IL-15 or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2170, SEQ ID NO: 3670, or SEQ ID NO: 3673.

220. The composition of any one of claims 212-214, 218, and 219, wherein the IL-15 or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2170, the sequence of SEQ ID NO: 3670, or the sequence of SEQ ID NO: 3673.

221. The composition of any one of claims 212-214 and 218-220, wherein the at least one cytokine molecule or a functional fragment or variant thereof further comprises an IL15Ralpha dimerizing domain covalently linked the IL-15 or a functional fragment or variant thereof.

222. The composition of claim 221, wherein the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or a functional fragment or variant thereof.

223. The composition of claim 221 or 222, wherein the IL15Ralpha dimerizing domain comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3472.WSGR Docket No.53676-767.601 224. The composition of any one of claims 221-223, wherein the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

225. The composition of any one of claims 221-224, wherein the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

226. The composition of any one of claims 221-225, wherein the IL-15 or a functional fragment or variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

227. The composition of any one of claims 212-214 and 218-226, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3474, SEQ ID NO: 3671, or SEQ ID NO: 3674.

228. The composition of any one of claims 212-214 and 218-227, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3474, the sequence of SEQ ID NO: 3671, or the sequence of SEQ ID NO: 3674.

229. The composition of any one of claims 211-214, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-7 or a functional fragment or variant thereof.

230. The composition of any one of claims 212-214 and 229, wherein the IL-7 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

231. The composition of any one of claims 212-214, 229, and 230, wherein the IL-7 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

232. The composition of any one of claims 211-214, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-12 or a functional fragment or variant or variant thereof.

233. The composition of any one of claims 212-214 and 232, wherein the IL-12 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

234. The composition of any one of claims 212-214, 232, and 233, wherein the IL-12 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3542, the sequence of SEQ ID NO: 3543, or the sequence of SEQ ID NO: 3545.

235. The composition of any one of claims 211-214, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-18 of a functional fragment or variant thereof.

236. The composition of any one of claims 212-214 and 235, wherein the IL-18 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2192.

237. The composition of any one of claims 212-214, 235, and 236, wherein the IL-18 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 2192.WSGR Docket No.53676-767.601 238. The composition of any one of claims 211-214, wherein the at least one cytokine molecule or a functional fragment or variant thereof comprises IL-21 or a functional fragment or variant thereof.

239. The composition of any one of claims 212-214 and 238, wherein the IL-21 or the functional fragment or variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3540.

240. The composition of any one of claims 212-214, 238, and 239, wherein the IL-21 or the functional fragment or variant thereof comprises the sequence of SEQ ID NO: 3540.

241. A recombinant polynucleotide molecule comprising a polynucleotide sequence encoding the agent of the composition of any one of claims 1-240.

242. The recombinant polynucleotide molecule of claim 241, wherein the recombinant polynucleotide molecule is an isolated recombinant polynucleotide molecule.

243. A vector comprising the recombinant polynucleotide molecule of claim 241 or 242.

244. A cell comprising the recombinant polynucleotide molecule of claim 241 or 242, or the vector of claim 243.

245. A method of making the agent of the composition of any one of claims 1-240, comprising culturing the cell of claim 244, under conditions suitable for expression of the agent.

246. A pharmaceutical composition comprising the composition of any one of claims 1-240, the recombinant polynucleotide molecule of claim 241 or 242, the vector of claim 243, or the cell of claim 244, and a pharmaceutically acceptable carrier, excipient, or diluent.

247. A method of treating a condition or disease in a subject in need thereof comprising administering to the subject a therapeutically effective amount of the composition of any one of claims 1-240, the recombinant polynucleotide molecule of claim 241 or 242, the vector of claim 243, or the cell of claim 244, or the pharmaceutical composition of claim 246, or any combination thereof, thereby treating the condition or disease in the subject.

248. The method of claim 247, wherein the condition or disease is cancer.

249. The method of claim 248, wherein the cancer is leukemia or lymphoma.

250. The method of claim 247 or 248, wherein the cancer is selected from the group consisting of acquired immune deficiency syndrome (AIDS)-associated lymphoma, Angioimmunoblastic T-cell lymphoma, Adult T-cell leukemia / lymphoma, Burkitt lymphoma, Central nervous system (CNS) lymphoma, Diffuse large B-cell lymphoma (DLBCL), Lymphoblastic lymphoma, Mantle cell lymphoma (MCL), Peripheral T-cell lymphoma (PTCL), Transformed follicular and transformed mucosa-associated lymphoid tissue (MALT) lymphomas, Cutaneous T-cell lymphoma (mycosis fungoides and Sézary syndrome), Follicular lymphoma, Lymphoplasmacytic lymphoma / Waldenström macroglobulinemia, Marginal zone B-cell lymphoma, Gastric mucosa- associated lymphoid tissue (MALT) lymphoma, Chronic lymphocytic leukemia / small-cell lymphocytic lymphoma (CLL / SLL), Extranodal T- / NK-cell lymphoma, and Anaplastic large-cell lymphoma.WSGR Docket No.53676-767.601 251. The method of claim 250, wherein peripheral T-cell lymphoma (PTCL) is Hepatosplenic T-cell lymphoma (HSGDTCL), Subcutaneous paniculitis-like T-cell lymphoma, or Enteropathy-associated T-cell lymphoma.

252. The method of claim 250, wherein the cutaneous T-cell lymphoma is mycosis fungoides and Sézary syndrome.

253. The method of claim 250, wherein the extranodal T- / NK-cell lymphoma is a nasal type.

254. The method of claim 250, wherein the anaplastic large-cell lymphoma is primary cutaneous anaplastic large-cell lymphoma or systemic anaplastic large-cell lymphoma.

255. The method of claim 248, wherein the cancer is a solid tumor.

256. The method of claim 255, wherein the solid tumor is selected from the group consisting of pancreatic cancer, breast cancer, colon cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, esophageal cancer, endometrial cancer, gastric cancer, head and neck cancer, kidney cancer, and prostate cancer.

257. The method of claim 256, wherein the pancreatic cancer is pancreatic adenocarcinoma or pancreatic ductal adenocarcinoma.

258. The method of claim 256, wherein the lung cancer is small cell lung cancer (SCLC) or non-small cell lung cancer (NSCLC).

259. The method of claim 247, wherein the condition or disease is an autoimmune disease.

260. The method of claim 259, wherein the autoimmune disease is selected from the group consisting of: amyotrophic lateral sclerosis (ALS), acute anterior uveitis (UAA), celiac disease (CD), ankylosing spondylitis (AS), Covid-induced multisystem inflammatory syndrome in children (MIS-C), primary Sjogren’s syndrome (PSS), Churg-Strauss syndrome, sarcoidosis, systemic lupus erythematosus (SLE), type 1 diabetes, autoimmune hepatitis (e.g., type 1 or type 2), type 1 narcolepsy, primary sclerosing cholangitis, primary biliary cirrhosis, multiple sclerosis, Guillain-Barre syndrome and the AMAN (axonal & neuronal neuropathy), chronic inflammatory demyelinating polyneuropathy (CIDP), transverse myelitis, Tolosa-Hunt syndrome (THS), Devic’s disease (neuromyelitis optica), paraneoplastic cerebellardegeneration (PCD), Lambert-Eaton syndrome, psoriasis, scleroderma, CREST (calcinosis,Raynaud phenomenon, esophageal dysmotility, sclerodactyly, and telangiectasia) syndrome, dermatitis herpetiformis, dermatomyositis, bullous pemphigoid, cicatricial pemphigoid / benign mucosal pemphigoid, pemphigoid gestationis, rheumatoid arthritis (RA), psoriatic arthritis, relapsing polychondritis, chronic recurrent multifocal osteomyelitis (CRMO), vasculitis, Kawasaki disease, joint implant failure, granulomatosis with polyangiitis (GPA), Behcet’s disease (vasculitis),Takayasu’s arteritis, polyarteritis nodosa, microscopic polyangiitis (MPA), leukocytoclasticvasculitis, Cogan’s syndrome, uveitis, peripheral uveitis (Pars planitis), scleritis, autoimmune inner ear disease (AIED), Crohn’s, ulcerative colitis (UC), Dressler’s syndrome, Rheumatic fever, Evans syndrome, paroxysmal nocturnal hemoglobinuria (PNH), hemolytic anemia, thrombocytopenic purpura (TTP), polymyositis, juvenile myositis (JM), including JuvenileWSGR Docket No.53676-767.601 Dermatomyositis (JDM) and Juvenile Polymyositis (JPM), ocular cicatricial pemphigoid, and Hashimoto’s thyroiditis.

261. The method of claim 260, wherein the autoimmune disease is multiple sclerosis, UAA, PsA, type I narcolepsy, ALS, Crohn’s disease, celiac disease, Type I diabetes, Sjögren’s syndrome, ankylosing spondylitis, joint implant failure, or COVID-induced multisystem inflammatory syndrome.

262. A method of expanding and / or activating NK cells in a NK cell population comprising contacting the NK cell population with an effective amount of the composition of any one of claims 1-240, the recombinant polynucleotide molecule of claim 241 or 242, the vector of claim 243, or the cell of claim 244, or the pharmaceutical composition of claim 246, or any combination thereof, thereby, expanding and / or activating the NK cells in the NK cell population.

263. The method of claim 262, wherein the expanding and / or activating NK cells is in vivo or ex vivo expansion and / or activation.

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