Multifunctional molecules binding to TCR and uses thereof

Abstract

Provides herein are compositions or formulations comprising a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region and a second agent comprising a second domain that binds to CD3, and methods of treating conditions or diseases in a subject using the same. Also provides herein are multifunctional molecules comprising T cell receptor variable beta-binding moieties, a tumor-associated antigen binding moiety, and cytokines, and methods of treating conditions or diseases in a subject using the same.

Description

WSGR Docket No.53676-769.601 MULTIFUNCTIONAL MOLECULES BINDING TO TCR AND USES THEREOF CROSS REFERENCE

[0001] This application claims the benefit of U.S. provisional Application No.63 / 579,761, filed on August 30, 2023; U.S. provisional Application No.63 / 579,741, filed on August 30, 2023; U.S. provisional Application No.63 / 588,390, filed on October 6, 2023; U.S. provisional Application No. 63 / 588,600, filed on October 6, 2023; and U.S. provisional Application No.63 / 601,442, filed on November 21, 2023; each of which is incorporated herein by reference in its entirety. BACKGROUND

[0002] Currently available molecules designed to redirect T cells to promote tumor cell lysis for cancer immunotherapy typically target the CD3 epsilon (CD3e) subunit of the T cell receptor (TCR). However, there are limitations to this approach. Previous studies have shown that some tumors do not respond well to anti-CD3e therapy alone. Low doses of anti-CD3e monoclonal antibody (mAb) can also cause T cell dysfunction and exert immunosuppressive effects. In addition, anti-CD3e mAbs bind to all T cells and thus activate a large number of T cells. Such non-physiological massive activation of T cells by these anti-CD3e mAbs can result in the production of proinflammatory cytokines such as IFN-gamma, IL-1- beta, IL-6, IL-10 and TNF-alpha, causing a “cytokine storm” known as the cytokine release syndrome (CRS), which is also associated with neurotoxicity (NT). Thus, there is a need for improved T cell receptor-binding molecules that redirect T cells for cancer immunotherapy. SUMMARY

[0003] In an aspect, provided herein is a composition for treating a disease or condition in a subject in need thereof comprising: (a) a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region; and (b) a second agent comprising a second domain that binds to CD3.

[0004] In another aspect, provided herein is a formulation for treating a disease or condition in a subject in need thereof comprising: (a) a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region; and (b) a second agent comprising a second domain that binds to CD3.

[0005] In some embodiments, the composition or the formulation comprises a therapeutically effective amount of the first agent and the second agent.

[0006] In some embodiments, the subject exhibits an improved therapeutic effect after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0007] In some embodiments, the subject comprises more activated and / or expanded T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.WSGR Docket No.53676-769.601

[0008] In some embodiments, the subject comprises less exhausted T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0009] In some embodiments, the subject exhibits more anti-tumor efficacy after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0010] In some embodiments, the subject comprises T cells that are more cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0011] In some embodiments, the subject comprises more T cells that are cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0012] In some embodiments, the subject exhibits more cytotoxic activity against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0013] In some embodiments, the TCR variable region is a TCR β variable (TCRβV) region.

[0014] In some embodiments, the TCR variable region is a human TCR β variable (TCRβV) region.

[0015] In some embodiments, the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of TCRβ V1 subfamily, TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 sub-family, TCRβ V6 subfamily, TCRβ V7 subfamily, TCRβ V8 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V14 sub-family, TCRβ V15 subfamily, TCRβ V16 subfamily, TCRβ V17 subfamily, TCRβ V18 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V22 subfamily TCRβ V23 subfamily, TCRβ V24 subfamily, TCRβ V25 subfamily, TCRβ V26 subfamily, TCRβ V27 subfamily, TCRβ V28 subfamily, TCRβ V29 subfamily, and TCRβ V30 subfamily.

[0016] In some embodiments, the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V1 subfamily comprising TCRβ V1*01; (ii) TCRβ V2 subfamily comprising one or more selected from TCRβ V2*01, TCRβ V2*02, and TCRβ V2*03; (iii) TCRβ V3 subfamily comprising one or more selected from TCRβ V3-1*01 and TCRβ V3-1*02; (iv) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1*01, TCRβ V4-1*02, TCRβ V4-2*01, TCRβ V4-2*02, TCRβ V4-3*01, TCRβ V4-3*02, TCRβ V4-3*03, and TCRβ V4-3*04; (v) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-1*01, TCRβ V5-1*02, TCRβ V5-3*01, TCRβ V5-3*02, TCRβ V5-4*01, TCRβ V5-4*02, TCRβ V5-4*03, TCRβ V5-4*04, TCRβ V5- 5*01, TCRβ V5-5*02, TCRβ V5-5*03, TCRβ V5-6*01, TCRβ V5-7*01, TCRβ V5-8*01, and TCRβ V5- 8*02;WSGR Docket No.53676-769.601 (vi) TCRβ V6 subfamily comprising one or more selected from TCRβ V6-1*01, TCRβ V6-2*01, TCRβ V6-3*01, TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-5*01, TCRβ V6-6*01, TCRβ V6-6*02, TCRβ V6- 6*03, TCRβ V6-6*04, TCRβ V6-6*05, TCRβ V6-7*01, TCRβ V6-8*01, and TCRβ V6-9*01; (vii) TCRβ V7 subfamily comprising one or more selected from TCRβ V7-1*01, TCRβ V7-2*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*04, TCRβ V7-3*01, TCRβ V7-3*02, TCRβ V7-3*03, TCRβ V7- 3*04, TCRβ V7-3*05, TCRβ V7-4*01, TCRβ V7-4*02, TCRβ V7-6*01, TCRβ V7-6*02, TCRβ V7- 7*01, TCRβ V7-7*02, TCRβ V7-8*01, TCRβ V7-8*02, TCRβ V7-8*03, TCRβ V7-9*01, TCRβ V7- 9*02, TCRβ V7-9*03, TCRβ V7-9*04, TCRβ V7-9*05, TCRβ V7-9*06, and TCRβ V7-9*07. (viii) TCRβ V8 subfamily comprising one or more selected from TCRβ V8-1*01, TCRβ V8-1*02, TCRβ V8-2*01, and TCRβ V8-2*02; (ix) TCRβ V9 subfamily comprising one or more selected from TCRβ V9-1*01, TCRβ V9-1*02, and TCRβ V9-1*03; (x) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-1*03, TCRβ V10-2*01, TCRβ V10-2*02, TCRβ V10-3*01, TCRβ V10-3*02, TCRβ V10-3*03, and TCRβ V10-3*04; (xi) TCRβ V11 subfamily comprising TCRβ V11-1*01, TCRβ V11-2*01, TCRβ V11-2*02, TCRβ V11- 2*03, TCRβ V11-3*01, TCRβ V11-3*02, TCRβ V11-3*03, and TCRβ V11-3*04; (xii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-3*01, TCRβ V12-4*01, TCRβ V12-4*02, and TCRβ V12-5*01; (xiii) TCRβ V13 subfamily comprising one or more selected from TCRβ V13*01 and TCRβ V13*02; (xiv) TCRβ V14 subfamily comprising one or more comprising from TCRβ V14*01 and TCRβ V14*02; (xv) TCRβ V15 subfamily comprising one or more selected from TCRβ V15*01, TCRβ V15*02, and TCRβ V15*03; (xvi) TCRβ V16 subfamily comprising one or more selected from TCRβ V16*01, TCRβ V16*02, and TCRβ V16*03; (xvii) TCRβ V17 subfamily comprising TCRβ V17*01; (xviii) TCRβ V18 subfamily comprising TCRβ V18*01; (xix) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01, TCRβ V19*02, and TCRβ V19*03; (xx) TCRβ V20 subfamily comprising one or more selected from TCRβ V20-1*01, TCRβ V20-1*02, TCRβ V20-1*03, TCRβ V20-1*04, TCRβ V20-1*05, TCRβ V20-1*06, and TCRβ V20-1*07; (xxi) TCRβ V21 subfamily comprising one or more selected from TCRβ V21-1*01 and TCRβ V21- 1*02; (xxii) TCRβ V22 subfamily comprising TCRβ V22-1*01; (xxiii) TCRβ V23 subfamily comprising TCRβ V23-1*01; (xxiv) TCRβ V24 subfamily comprising TCRβ V24-1*01; (xxv) TCRβ V25 subfamily comprising TCRβ V25-1*01; (xxvi) TCRβ V26 subfamily comprising TCRβ V26-1*01;WSGR Docket No.53676-769.601 (xxvii) TCRβ V27 subfamily comprising TCRβ V27*01; (xxviii) TCRβ V28 subfamily comprising TCRβ V28*01; (xxix) TCRβ V29 subfamily comprising one or more selected from TCRβ V29-1*01, TCRβ V29-1*02, and TCRβ V29-1*03; and (xxx) TCRβ V30 subfamily comprising one or more selected from TCRβ V30*01, TCRβ V30*02, TCRβ V30*03, TCRβ V30*04, and TCRβ V30*05.

[0017] In some embodiments, the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRβV region.

[0018] In some embodiments, the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', an F(ab')2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

[0019] In some embodiments, the TCR variable region is a TCR α variable region.

[0020] In some embodiments, the TCR variable region is a human TCR α variable region.

[0021] In some embodiments, the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

[0022] In some embodiments, the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02; (ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02; (iii) TCRα V3 subfamily comprising TCRα V3*01; (iv) TCRα V4 subfamily comprising TCRα V4*01; (v) TCRα V5 subfamily comprising TCRα V5*01 (vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06; (vii) TCRα V7 subfamily comprising TCRα V7*01; (viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8-4*01, TCRαWSGR Docket No.53676-769.601 V8-4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8-6*02, and TCRα V8-7*01; (ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; (x) TCRα V10 subfamily comprising TCRα V10*01; (xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02; (xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02; (xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04; (xiv) TCRα V16 subfamily comprising TCRα V16*01; (xv) TCRα V17 subfamily comprising TCRα V17*01; (xvi) TCRα V18 subfamily comprising TCRα V18*01; (xvii) TCRα V19 subfamily comprising TCRα V19*01 (xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04; (xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02; (xx) TCRα V22 subfamily comprising TCRα V22*01; (xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04; (xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02; (xxiii) TCRα V25 subfamily comprising TCRα V25*01; (xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRα V26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02; (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01; (xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02; (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38-1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01; (xxxiii) TCRα V40 subfamily comprising TCRα V40*01; andWSGR Docket No.53676-769.601 (xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

[0023] In some embodiments, the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRαV region.

[0024] In some embodiments, the first agent comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or fragment thereof, IgG2 heavy chain constant region or fragment thereof, IgG3 heavy chain constant region or fragment thereof, IgGA1 heavy chain constant region or fragment thereof, IgGA2 heavy chain constant region or fragment thereof, IgG4 heavy chain constant region or fragment thereof, IgJ heavy chain constant region or fragment thereof, IgM heavy chain constant region or fragment thereof, IgD heavy chain constant region or fragment thereof, and IgE heavy chain constant region or fragment thereof.

[0025] In some embodiments, the first agent comprises a kappa light chain constant region or fragment thereof, a lambda light chain constant region or fragment thereof, or a combination thereof.

[0026] In some embodiments, the first agent is a multifunctional molecule.

[0027] In some embodiments, the first agent comprises at least two non-contiguous polypeptide chains; wherein a first polypeptide chain of the at least two non-contiguous polypeptide chains comprises a first Fc region, and a second polypeptide chain of the at least two non-contiguous polypeptide chains comprises a second Fc region.

[0028] In some embodiments, the first Fc region and the second Fc region comprise an Fc interface with a knob-in-a hole.

[0029] In some embodiments, the first Fc region, the second Fc region, or a combination thereof comprises an Asn297Ala mutation, a Leu234Ala / Leu235Ala mutation, or a combination thereof.

[0030] In some embodiments, the first agent is a multispecific molecule that further comprises one or more of a tumor-targeting moiety, a cytokine molecule, a stromal modifying moiety, or an immune cell engager.

[0031] In some embodiments, the first agent is a multispecific molecule that further comprises a cytokine molecule.

[0032] In some embodiments, the first agent comprises the following configuration: A-, B-[dimerization module]-C, -D, wherein: (a) the dimerization module comprises a first immunoglobulin chain constant region and a second immunoglobulin chain constant region; (b) A and C are linked to the first immunoglobulin chain constant region; and B and D are linked to the second immunoglobulin chain constant region; (c) A, B, C, and D are independently (i) absent; (ii) the first binder; (iii) the tumor targeting moiety, (iv) the cytokine molecule; (v) stromal modifying moiety; or (vi) the immune cell engager; and wherein at least one, two, or three of A, B, C, and D comprises the first agent.WSGR Docket No.53676-769.601

[0033] In some embodiments, the first immunoglobulin chain constant region comprises the first fragment crystallizable region (Fc region) and the second first immunoglobulin chain constant region comprises the second fragment crystallizable region (Fc region).

[0034] In some embodiments, dimerization of the first Fc region and the second Fc region is enhanced by providing an Fc interface of the first Fc region and the second Fc region with one or more of a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms relative to a non-engineered interface.

[0035] 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, a macrophage cell engager, and any combination thereof.

[0036] In some embodiments, the cytokine molecule is selected from the group consisting of inter- leukin-2 (IL-2) or functional variant thereof, interleukin-7 (IL-7) or functional variant thereof, in- terleukin-12 (IL-12) or functional variant thereof, interleukin-15 (IL-15) or functional variant thereof, interleukin-18 (IL-18) or functional variant thereof, interleukin-21 (IL-21) or functional variant thereof, interferon gamma or functional variant thereof, and any combination thereof.

[0037] In some embodiments, the cytokine molecule comprises interleukin-2 (IL-2) or functional variant thereof.

[0038] In some embodiments, the interleukin-2 (IL-2) or functional variant thereof comprises the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

[0039] In some embodiments, the cytokine molecule comprises interleukin-15 (IL-15) or functional variant thereof.

[0040] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof comprises the sequence of SEQ ID NO: 2170.

[0041] In some embodiments, the cytokine molecule further comprises an IL15Ralpha dimerizing domain covalently linked the interleukin-15 (IL-15) or functional variant thereof.

[0042] In some embodiments, the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or functional variant thereof.

[0043] In some embodiments, the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

[0044] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

[0045] In some embodiments, the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

[0046] In some embodiments, the cytokine molecule comprises the sequence of SEQ ID NO: 3474.

[0047] In some embodiments, the second domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to CD3.WSGR Docket No.53676-769.601

[0048] In some embodiments, the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', an F(ab')2, an F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

[0049] In some embodiments, the second domain binds to CD3ε. In some embodiments, the second domain binds to human CD3ε.

[0050] In some embodiments, the second domain is selected from the group consisting of muromonab- CD3, an antigen binding domain thereof, or a functional fragment or variant thereof, otelixizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, teplizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, and visilizumab, an antigen binding domain thereof, or a functional fragment or variant thereof.

[0051] In some embodiments, the second agent further comprises a full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen.

[0052] In some embodiments, the full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen is operatively linked to the second domain.

[0053] In some embodiments, the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, TRP-1 / -2, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, MUC-1, CA- 125, BAGE, GAGE, NY-ESO-1, β-catenin, CDK4, CDC27, α actinin-4, TRP1 / gp75, TRP2, gp100, Melan-A / MART1, gangliosides, WT1, EphA3, Epidermal growth factor receptor (EGFR), MART-2, MART-1, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

[0054] In some embodiments, the condition or disease is cancer. In some embodiments, the cancer is a solid tumor, a hematological cancer, a metastatic cancer, a soft tissue tumor, or a combination thereof.

[0055] In some embodiments, the cancer is the solid tumor, and wherein the solid tumor is selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

[0056] In some embodiments, the cancer is the hematological cancer, and wherein the hematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lympho-ma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T- cell lymphoma, acute lymphocytic leukemia, and a combination thereof.WSGR Docket No.53676-769.601

[0057] In some embodiments, the Non-Hodgkin’s lymphoma is selected from the group consisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

[0058] In some embodiments, the T-cell lymphoma is peripheral T-cell lymphoma.

[0059] In some embodiments, the cancer is characterized by a cancer antigen present on the cancer.

[0060] In some embodiments, the cancer antigen is a tumor antigen, a stromal antigen, or a hematological antigen.

[0061] In some embodiments, the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, ganglioside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, TRP1 / gp75, TRP2, gangliosides, WT1, Epi-dermal growth factor receptor (EGFR), MART-2, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Integrins, Carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

[0062] In some embodiments, the composition is pharmaceutical composition.

[0063] In some embodiments, the formulation is pharmaceutical formulation.

[0064] In some embodiments, the composition or formulation further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

[0065] In another aspect, provided herein is a method of treating a disease or condition in a subject in need thereof comprising: (a) administering a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region; and (b) administering a second agent comprising a second domain that binds to CD3; wherein a therapeutically effective amount of the first agent and the second agent is administered to the subject, thereby treating the disease or condition in the subject.

[0066] In another aspect, provided herein is a method of treating a disease or condition in a subject in need thereof comprising administering to the subject a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, wherein the subject has been previously treated with a second agent comprising a second domain that binds to CD3, thereby treating the disease or condition in the subject.

[0067] In another aspect, provided herein is a method of improving a therapeutic efficacy of a second agent comprising a second domain that binds to CD3 in a subject in need thereof comprising:WSGR Docket No.53676-769.601 administering a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, thereby improving the therapeutic efficacy of the second agent in the subject.

[0068] In another aspect, provided herein is a combination therapy for treating a disease or condition in a subject in need thereof a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, and a second agent comprising a second domain that binds to CD3.

[0069] In some embodiments, administering the first agent and the second agent induces an improved therapeutic effect in the subject, relative to a subject administered the second agent without the first agent.

[0070] In some embodiments, administering the first agent and the second agent induces more activated and / or expanded T cells in the subject, relative to a subject administered the second agent without the first agent.

[0071] In some embodiments, administering the first agent and the second agent induces less exhausted T cells in the subject, relative to a subject administered the second agent without the first agent.

[0072] In some embodiments, administering the first agent and the second agent induces more anti-tumor efficacy in the subject, relative to a subject administered the second agent without the first agent.

[0073] In some embodiments, administering the first agent and the second agent induces more cytotoxicity against tumor cells or cancer cells in the subject, relative to a subject administered the second agent without the first agent.

[0074] In some embodiments, administering the first agent and the second agent expands or activates more T cells that are cytotoxic against tumor cells or cancer cells in the subject, relative to a subject administered the second agent without the first agent.

[0075] In some embodiments, administering the first agent and the second agent induces more cytotoxic activity against tumor cells or cancer cells in the subject, relative to a subject administered the second agent without the first agent.

[0076] In some embodiments, the first agent and the second agent are not concomitantly administered to the subject.

[0077] In some embodiments, the first agent and the second agent are concomitantly administered to the subject.

[0078] In some embodiments, the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years after the subject has been treated with a dose of the second agent.

[0079] In some embodiments, the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years after the subject has been treated with a first dose of the second agent.

[0080] In some embodiments, the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks,WSGR Docket No.53676-769.601 at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years after the subject has been treated with a last dose of the second agent.

[0081] In some embodiments, the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with the second agent.

[0082] In some embodiments, the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with a dose of the second agent.

[0083] In some embodiments, the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with a first dose of the second agent.

[0084] In some embodiments, the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with a last dose of the second agent.

[0085] In some embodiments, the method comprises administering the first agent to the subject from 1 hour to 24 hours after being treated with a dose of the second agent.

[0086] In some embodiments, the method comprises administering the first agent to the subject from 1 day to 7 days after being treated with a dose of the second agent.

[0087] In some embodiments, the method comprises administering the first agent to the subject from 1 week to 4 weeks after being treated with a dose of the second agent.

[0088] In some embodiments, the method comprises administering the first agent to the subject from 1 month to 12 months after being treated with a dose of the second agent.

[0089] In some embodiments, the method comprises administering the first agent to the subject from 1 year to 3 years after being treated with a dose of the second agent.

[0090] In some embodiments, the method comprises administering the first agent to the subject from 1 hour to 24 hours after being treated with a first dose of the second agent.

[0091] In some embodiments, the method comprises administering the first agent to the subject from 1 day to 7 days after being treated with a first dose of the second agent.

[0092] In some embodiments, the method comprises administering the first agent to the subject from 1 week to 4 weeks after being treated with a first dose of the second agent.

[0093] In some embodiments, the method comprises administering the first agent to the subject from 1 month to 12 months after being treated with a first dose of the second agent.

[0094] In some embodiments, the method comprises administering the first agent to the subject from 1 year to 3 years after being treated with a first dose of the second agent.WSGR Docket No.53676-769.601

[0095] In some embodiments, the method comprises administering the first agent to the subject from 1 hour to 24 hours after being treated with a last dose of the second agent.

[0096] In some embodiments, the method comprises administering the first agent to the subject from 1 week to 4 weeks after being treated with a last dose of the second agent.

[0097] In some embodiments, the method comprises administering the first agent to the subject from 1 month to 12 months after being treated with a last dose of the second agent.

[0098] In some embodiments, the method comprises administering the first agent to the subject from 1 year to 3 years after being treated with a last dose of the second agent.

[0099] In some embodiments, the method comprises administering the first agent to the subject on the same day the subject had been previously treated with the second agent.

[0100] In some embodiments, the method is repeated twice or more.

[0101] In some embodiments, the administration of the first agent comprises one or more doses.

[0102] In some embodiments, the administration of the first agent comprises two or more doses.

[0103] In some embodiments, the administration of the first agent comprises one dose per week.

[0104] In some embodiments, the administration of the first agent comprises two or more doses per week.

[0105] In some embodiments, the first agent is administered to the subject once every two weeks.

[0106] In some embodiments, the first agent is administered to the subject once every three weeks.

[0107] In some embodiments, the first agent is administered to the subject once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0108] In some embodiments, the administration of the first agent comprises two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0109] In some embodiments, the first agent is administered to the subject once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0110] In some embodiments, the first agent is administered to the subject once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0111] In some embodiments, the administration of the second agent comprises one or more doses.

[0112] In some embodiments, the administration of the second agent comprises two or more doses.

[0113] In some embodiments, the administration of the second agent comprises one dose per week.

[0114] In some embodiments, the administration of the second agent comprises two or more doses per week.

[0115] In some embodiments, the second agent is administered to the subject once every two weeks.

[0116] In some embodiments, the second agent is administered to the subject once every three weeks.

[0117] In some embodiments, the second agent is administered to the subject once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.WSGR Docket No.53676-769.601

[0118] In some embodiments, the administration of the second agent comprises two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0119] In some embodiments, the second agent is administered to the subject once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0120] In some embodiments, the second agent is administered to the subject once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0121] In some embodiments, the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years prior to administering the second agent to the subject.

[0122] In some embodiments, the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years prior to administering the second agent to the subject.

[0123] In some embodiments, the method comprises administering the first agent to the subject from 1 hour to 24 hours prior to administering the second agent to the subject.

[0124] In some embodiments, the method comprises administering the first agent to the subject from 1 day to 7 days prior to administering the second agent to the subject.

[0125] In some embodiments, the method comprises administering the first agent to the subject from 1 week to 4 weeks prior to administering the second agent to the subject.

[0126] In some embodiments, the method comprises administering the first agent to the subject from 1 month to 12 months prior to administering the second agent to the subject.

[0127] In some embodiments, the method comprises administering the first agent to the subject from 1 year to 3 years prior to administering the second agent to the subject.

[0128] In some embodiments, the administration of the first agent comprises two or more doses, and each dose comprises a same amount of the first agent.

[0129] In some embodiments, the administration of the first agent comprises two or more doses, and each dose comprises different amounts of the first agent.

[0130] In some embodiments, the administration of the second agent comprises two or more doses, and each dose comprises a same amount of the second agent.

[0131] In some embodiments, the administration of the second agent comprises two or more doses, and each dose comprises different amounts of the second agent.

[0132] In some embodiments, the first agent is administered to the subject at a dose of from about 0.001 mg / kg to about 20 mg / kg.

[0133] In some embodiments, the second agent is administered to the subject at a dose of from about 0.001 mg / kg to about 20 mg / kg.WSGR Docket No.53676-769.601

[0134] In some embodiments, the method comprises administering the first agent and the second agent to the subject simultaneously or on the same day.

[0135] In some embodiments, the disease or condition in the subject is treated to a higher extent relative to a corresponding method in which the first agent is administered and the second agent is not administered.

[0136] In some embodiments, the method is more effective to treat the disease or condition in the subject relative to a method in which the first agent is administered to the subject but not the second agent.

[0137] In some embodiments, the method is more effective to treat the disease or condition in the subject relative to a method in which the second agent is administered to the subject but not the first agent.

[0138] In some embodiments, the first agent is administered to the subject prior to, concurrently, or after administration of the second agent to the subject.

[0139] In another aspect, provided herein is a kit comprising: (i) a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, (ii) a second agent comprising a second domain that binds to CD3, and (iii) instructions for administering the first agent and the second agent to a human subject in need thereof to treat a disease or condition in the human subject.

[0140] In some embodiments, the instructions comprise instructions for administering a therapeutic effective amount of the first agent to the human subject.

[0141] In some embodiments, the instructions comprise instructions for administering a therapeutic effective amount of the second agent to the human subject.

[0142] In some embodiments, the instructions comprise instructions for frequency of administering the first agent to the human subject.

[0143] In some embodiments, the instructions comprise instructions for frequency of administering the second agent to the human subject.

[0144] In some embodiments, the instructions comprise instructions to administer the second agent to the human subject prior to administering the first agent to the human subject.

[0145] In some embodiments, the instructions comprise instructions to administer the second agent and the first agent not concomitantly to the human subject.

[0146] In some embodiments, the instructions comprise instructions to administer to the human subject one or more doses of the second agent.

[0147] In some embodiments, the instructions comprise instructions to administer to the human subject two or more doses of the second agent.

[0148] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent one dose per week.

[0149] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent two or more doses per week.

[0150] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent once every two weeks.WSGR Docket No.53676-769.601

[0151] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent once every three weeks.

[0152] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0153] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0154] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0155] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0156] In some embodiments, the instructions comprise instructions to administer one or more doses of the first agent.

[0157] In some embodiments, the instructions comprise instructions to administer two or more doses of the first agent.

[0158] In some embodiments, the administration of the first agent comprises one dose per week.

[0159] In some embodiments, the administration of the first agent comprises two or more doses per week.

[0160] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent once every two weeks.

[0161] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent once every three weeks.

[0162] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0163] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0164] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

[0165] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.WSGR Docket No.53676-769.601

[0166] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years prior to administering the first agent to the subject.

[0167] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years prior to administering the first agent to the subject.

[0168] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent from 1 hour to 24 hours prior to administering the first agent to the subject.

[0169] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent from 1 day to 7 days prior to administering the first agent to the subject.

[0170] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent from 1 week to 4 weeks prior to administering the first agent to the subject.

[0171] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent to the subject from 1 month to 12 months prior to administering the first agent to the subject.

[0172] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent to the subject from 1 year to 3 years prior to administering the first agent to the subject.

[0173] In some embodiments, the instructions comprise instructions to administer to the human subject two or more doses of the second agent, and each dose comprises a same amount of the second agent.

[0174] In some embodiments, the instructions comprise instructions to administer to the human subject two or more doses of the second agent, and each dose comprises different amounts of the second agent.

[0175] In some embodiments, the instructions comprise instructions to administer to the human subject two or more doses of the first agent, and each dose comprises a same amount of the first agent.

[0176] In some embodiments, the instructions comprise instructions to administer to the human subject two or more doses of the first agent, and each dose comprises different amounts of the first agent.

[0177] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent at a dose of from about 0.001 mg / kg to about 20 mg / kg.

[0178] In some embodiments, the instructions comprise instructions to administer to the human subject the first agent at a dose of from about 0.001 mg / kg to about 20 mg / kg.

[0179] In some embodiments, the instructions comprise instructions to administer to the human subject the second agent and the first agent to the subject simultaneously or on the same day.

[0180] In some embodiments, the instructions comprise instructions to administer the first agent to the subject prior to, concurrently, or after administration of the second agent to the subject.

[0181] In some embodiments, the subject exhibits an improved therapeutic effect after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.WSGR Docket No.53676-769.601

[0182] In some embodiments, the subject comprises more activated and / or expanded T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0183] In some embodiments, the subject comprises less exhausted T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0184] In some embodiments, the subject exhibits more anti-tumor efficacy after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0185] In some embodiments, the subject comprises T cells that are more cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0186] In some embodiments, the subject comprises more T cells that are cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0187] In some embodiments, the subject exhibits more cytotoxic activity against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

[0188] In another aspect, provided herein is the use of the composition as provided herein, the formulation as provided herein, or the kit as provided herein for the method as provided herein or the combination therapy as provided herein.

[0189] In an aspect, provided herein is a multifunctional molecule comprising: (a) a tumor-associated antigen binding moiety, wherein the tumor-associated antigen is DLL-3 or CLDN18.2; and (b) a TCR (T Cell Receptor) variable region (TCRV)-binding moiety.

[0190] In some embodiments, the TCRV-binding moiety binds to a TCR β variable (TCRβV) region.

[0191] In some embodiments, the TCRV-binding moiety binds to a TCR α variable (TCRαV) region.

[0192] In some embodiments, the multifunctional molecule as provided herein further comprises at least one cytokine molecule or functional fragment or functional variant thereof.

[0193] In some embodiments, the TCRV-binding moiety is covalently linked to the at least one cytokine molecule or functional fragment or functional variant thereof.

[0194] In some embodiments, the multifunctional molecule comprises a first polypeptide comprising a first portion of a dimerization module, and a second polypeptide comprising a second portion of the dimerization module; wherein the first polypeptide and the second polypeptide are non-contiguous, and wherein the tumor-associated antigen binding moiety is linked to the first portion of the dimerization module, and the at least one cytokine molecule or functional fragment or functional variant thereof is linked to the first portion of the dimerization module, the second portion of the dimerization module, or a combination thereof.WSGR Docket No.53676-769.601

[0195] In some embodiments, (i) the tumor-associated antigen binding moiety is linked to the N-terminus of the first portion of the dimerization module, and the at least one cytokine molecule or functional fragment or functional variant thereof is linked to the C-terminus of the first portion of the dimerization module, the N-terminus of the second portion of the dimerization module, the C-terminus of the second portion of the dimerization module, or any combination thereof; or (ii) the tumor-associated antigen binding moiety is linked to the C-terminus of the first portion of the dimerization module, and the at least one cytokine molecule or functional fragment or functional variant thereof is linked to the N-terminus of the first portion of the dimerization module, the N-terminus of the second portion of the dimerization module, the C-terminus of the second portion of the dimerization module, or any combination thereof.

[0196] In some embodiments, the TCRV-binding moiety and the at least one cytokine molecule or functional fragment or functional variant thereof is within a single contiguous polypeptide chain of the first polypeptide or the second polypeptide.

[0197] In some embodiments, the tumor-associated antigen binding moiety, the TCRV-binding moiety, or a combination thereof comprises an antibody or antigen binding fragment thereof, wherein the antigen binding fragment comprises any one selected from the group consisting of a Fab, a F(ab')2, an Fv, a single chain Fv (scFv), a single domain antibody, a diabody, a camelid antibody, and a combination thereof.

[0198] In some embodiments, the TCRV-binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), or a single domain antibody.

[0199] In some embodiments, the TCRV-binding moiety comprises a first portion of the TCRV-binding moiety, and wherein the multifunctional molecule further comprises a third polypeptide comprising a second portion of the TCRV-binding moiety, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide.

[0200] In some embodiments, the first portion of the TCRV-binding moiety comprises a VH of the TCRV-binding moiety and the second portion of the TCRV-binding moiety comprises a VL of the TCRV-binding moiety, or the first portion of the TCRV-binding moiety comprises a VL of the TCRV- binding moiety and the second portion of the TCRV-binding moiety comprises a VH of the TCRV- binding moiety.

[0201] In some embodiments, the tumor-associated antigen binding moiety comprises a VH and a VL, or a single domain antibody.

[0202] In some embodiments, the tumor-associated antigen binding moiety comprises a first portion of the tumor-associated antigen binding moiety, and wherein the multifunctional molecule further comprises a fourth polypeptide comprising a second portion of the tumor-associated antigen binding moiety, wherein the fourth polypeptide is non-contiguous with the first polypeptide, the second polypeptide, and the third polypeptide.

[0203] In some embodiments, the first portion of the tumor-associated antigen binding moiety comprises a VH of the tumor-associated antigen binding moiety and the second portion of the tumor-associated antigen binding moiety comprises a VL of the tumor-associated antigen binding moiety, or the firstWSGR Docket No.53676-769.601 portion of the tumor-associated antigen binding moiety comprises a VL of the tumor-associated antigen binding moiety and the second portion of the tumor-associated antigen binding moiety comprises a VH of the tumor-associated antigen binding moiety.

[0204] In some embodiments, the first portion of the dimerization module and the second portion of the dimerization module are dimerized.

[0205] In some embodiments, (i) the tumor-associated antigen binding moiety further comprises a heavy chain constant domain 1 (CH1) linked to the VH of the tumor-associated antigen binding moiety; (ii) the TCRV-binding moiety further comprises a heavy chain constant domain 1 (CH1) linked to the VH of the TCRV-binding moiety; or (iii) a combination thereof.

[0206] In some embodiments, (i) the tumor-associated antigen binding moiety further comprises a light chain constant domain (CL) linked to the VL of the tumor-associated antigen binding moiety; (ii) the TCRV-binding moiety further comprises a light chain constant domain (CL) linked to the VL of the TCRV-binding moiety; or (iii) a combination thereof.

[0207] In some embodiments, (i) the CL linked to the VL of the tumor-associated antigen binding moiety comprises a kappa chain constant domain or a lambda chain constant domain; (ii) the CL linked to the VL of the TCRV-binding moiety comprises a kappa chain constant domain or a lambda chain constant domain; or (iii) a combination thereof.

[0208] In some embodiments, the kappa chain constant domain or the lambda chain constant domain comprises any one of the light chain constant region sequences listed in Table 3.

[0209] In some embodiments, the multifunctional molecule as provided herein further comprises: (i) a linker between the first portion of the dimerization module and the tumor-associated antigen binding moiety or the first portion of the tumor-associated antigen; (ii) a linker between the at least one cytokine molecule or functional fragment or functional variant thereof and the first portion of the dimerization module, a linker between the at least one cytokine molecule or functional fragment or functional variant thereof and the second portion of the dimerization module, or a combination thereof; (iii) a linker between the at least one cytokine molecule or functional fragment or functional variant thereof and the TCRV-binding moiety or the first portion of the TCRV-binding moiety; (iv) a linker between the VH and the VL of the tumor-associated antigen binding moiety; (v) a linker between the VH and the VL of the TCRV-binding moiety; (vi) a linker between the CH1 and the VH of the tumor-associated antigen binding moiety; (vii) a linker between the CH1 and the VH of the TCRV-binding moiety; (viii) a linker between the CL and the VL of the tumor-associated antigen binding moiety; (ix) a linker between the CL and the VL of the TCRV-binding moiety; or (x) any combination thereof.

[0210] 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.

[0211] In some embodiments, the linker is the peptide linker, and wherein the linker comprises the sequence of SEQ ID NO: 3308 or SEQ ID NO: 3643.WSGR Docket No.53676-769.601

[0212] In some embodiments, the multifunctional molecule is an isolated multifunctional molecule.

[0213] In some embodiments, the tumor-associated antigen binding moiety, the TCRV-binding moiety, or a combination thereof comprises the Fab or the scFv.

[0214] In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof is selected from the group consisting of interleukin-2 (IL-2) or functional fragment or functional variant thereof, interleukin-7 (IL-7) or functional fragment or functional variant thereof, interleukin-12 (IL-12) or functional fragment or functional variant thereof, interleukin-15 (IL-15) or functional fragment or functional variant thereof, interleukin-18 (IL-18) or functional fragment or functional variant thereof, interleukin-21 (IL-21) or functional fragment or functional variant thereof, or interferon gamma or functional fragment or functional variant thereof, or a combination thereof.

[0215] In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof comprises interleukin-2 (IL-2) or functional fragment or functional variant thereof.

[0216] In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof is an IL-2 variant comprising a substitution mutation.

[0217] In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof is an IL-2 variant comprising C125A mutation.

[0218] In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2270.

[0219] In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof comprises the sequence of SEQ ID NO: 2270.

[0220] In some embodiments, the first portion of the dimerization module comprises a first immunoglobulin constant regions (Fc regions) and the second portion of the dimerization module comprises a second Fc region.

[0221] In some embodiments, the first Fc region, the second Fc region, or a combination thereof is selected from the group consisting of an IgG1 Fc region or a fragment thereof, an IgG2 Fc region or a fragment thereof, an IgG3 Fc region or a fragment thereof, an IgGA1 Fc region or a fragment thereof, an IgGA2 Fc region or a fragment thereof, an IgG4 Fc region or a fragment thereof, an IgJ Fc region or a fragment thereof, an IgM Fc region or a fragment thereof, an IgD Fc region or a fragment thereof, and an IgE Fc region or a fragment thereof.

[0222] In some embodiments, the first Fc region, the second Fc region, or a combination thereof is selected from the group consisting of a human IgG1 Fc region or a fragment thereof, a human IgG2 Fc region or a fragment thereof, and a human IgG4 Fc region or a fragment thereof.

[0223] 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.WSGR Docket No.53676-769.601

[0224] In some embodiments, the first Fc region, the second Fc region, or a combination thereof comprises an amino acid substitution listed in Table 14. In some embodiments, the first Fc region, the second Fc region, or a combination thereof comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation.

[0225] In some embodiments, the first Fc region, the second Fc region, or a combination thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 3452, SEQ ID NO: 3454, SEQ ID NO: 3645, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO:3648, SEQ ID NO: 3649, SEQ ID NO: 3792, or SEQ ID NO: 3794.

[0226] In some embodiments, the first Fc region, the second Fc region, or a combination thereof comprises the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 3452, SEQ ID NO: 3454, SEQ ID NO: 3645, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO:3648, SEQ ID NO: 3649, SEQ ID NO: 3792, or SEQ ID NO: 3794.

[0227] In some embodiments, the first Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3792, and the second Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, or the first Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, and the second Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794.

[0228] In some embodiments, the first Fc region comprises a sequence having the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3792, and the second Fc region comprises the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, or the first Fc region comprises the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, and the second Fc region comprises the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794.

[0229] In some embodiments, the TCRV-binding moiety binds to one or more of a TCRβV subfamily.

[0230] In some embodiments, the TCRV-binding moiety binds to one or more of a TCRβV subfamily selected from the group consisting of TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 subfamily, TCRβ V6 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V16 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V23 subfamily, TCRβ V27 subfamily, and TCRβ V28 subfamily.

[0231] In some embodiments, the TCRV-binding moiety binds to one or more of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V2 subfamily comprising TCRβ V2*01; (ii) TCRβ V3 subfamily comprising TCRβ V3-1*01; (iii) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1, TCRβ V4-2, and TCRβ V4- 3; (iv) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-6*01, TCRβ V5-4*01, TCRβ V5-1*01, and TCRβ V5-8*01;WSGR Docket No.53676-769.601 (v) TCRβ V6 subfamily comprising one or more selected from 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, and TCRβ V6-1*01; (vi) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-3*01, and TCRβ V10-2*01; (vii) TCRβ V11 subfamily comprising TCRβ V11-2; (viii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-4*01, TCRβ V12-3*01, and TCRβ V12-5*01; (ix) TCRβ V13 subfamily comprising TCRβ V13*01; (x) TCRβ V16 subfamily comprising TCRβ V16*01; (xi) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01 and TCRβ V19*02; or (xii) TCRβ V20 subfamily comprising TCRβ V20-1*01, or TCRβ V20-1*02.

[0232] In some embodiments, the TCRV-binding moiety binds to TCRβ V6 subfamily or TCRβ V20 subfamily.

[0233] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

[0234] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

[0235] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

[0236] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.WSGR Docket No.53676-769.601

[0237] In some embodiments, the TCRV-binding moiety binds to one or more of a TCRαV subfamily.

[0238] In some embodiments, the TCRV-binding moiety binds to one or more of a one or more of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

[0239] In some embodiments, the TCRV-binding moiety binds to one or more of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02; (ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02; (iii) TCRα V3 subfamily comprising TCRα V3*01; (iv) TCRα V4 subfamily comprising TCRα V4*01; (v) TCRα V5 subfamily comprising TCRα V5*01; (vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06; (vii) TCRα V7 subfamily comprising TCRα V7*01; (viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8-4*01, TCRα V8- 4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8- 6*01, TCRα V8-6*02, and TCRα V8-7*01; (ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; (x) TCRα V10 subfamily comprising TCRα V10*01; (xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02; (xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02; (xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04; (xiv) TCRα V16 subfamily comprising TCRα V16*01; (xv) TCRα V17 subfamily comprising TCRα V17*01; (xvi) TCRα V18 subfamily comprising TCRα V18*01;WSGR Docket No.53676-769.601 (xvii) TCRα V19 subfamily comprising TCRα V19*01; (xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04; (xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02; (xx) TCRα V22 subfamily comprising TCRα V22*01; (xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04; (xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02; (xxiii) TCRα V25 subfamily comprising TCRα V25*01; (xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRα V26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02; (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01; (xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02; (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38-1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01; (xxxiii) TCRα V40 subfamily comprising TCRα V40*01; and (xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

[0240] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Table 23; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Table 23; or (iii) a combination thereof.

[0241] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Table 23; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Table 23; or (iii) a combination thereof.

[0242] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Table 23; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Table 23; or (iii) a combination thereof.WSGR Docket No.53676-769.601

[0243] In some embodiments, the TCRV-binding moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Table 23; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Table 23; or (iii) a combination thereof.

[0244] In some embodiments, the multifunctional molecule consists of a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises the tumor-associated antigen binding moiety linked to an Fc region, wherein the tumor-associated antigen binding moiety is an scFv that binds to a tumor-associated antigen.

[0245] In some embodiments, the multifunctional molecule consists of a first polypeptide, a second polypeptide, and a third polypeptide, wherein the first polypeptide chain comprises a first portion of the tumor-associated antigen binding moiety linked to an Fc region, wherein the tumor-associated antigen binding moiety is a Fab that binds to a tumor-associated antigen; and the third polypeptide comprises a second portion of the tumor-associated antigen binding moiety.

[0246] In some embodiments, the tumor-associated antigen binding moiety is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to DLL3.

[0247] In some embodiments, the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

[0248] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 34 or 35; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 34 or 35; or (iii) a combination thereof.

[0249] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 34 or 35; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 34 or 35; or (iii) a combination there of.

[0250] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 34 or 35; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 34 or 35; or (iii) a combination thereof.

[0251] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 34 or 35; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 34 or 35; or (iii) a combination thereof.WSGR Docket No.53676-769.601

[0252] In some embodiments, the tumor-associated antigen binding moiety is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to CLDN18.2.

[0253] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 38 or 39; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 38 or 39; or (iii) a combination thereof.

[0254] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 38 or 39; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 38 or 39; or (iii) a combination there of.

[0255] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 38 or 39; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 38 or 39; or (iii) a combination thereof.

[0256] In some embodiments, the tumor-targeting moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 38 or 39; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 38 or 39; or (iii) a combination thereof.

[0257] In another aspect, provided herein is a pharmaceutical composition comprising the multifunctional molecule as provided herein, the polynucleotide as provided herein, the vector as provided herein, or the cell as provided herein, and a pharmaceutically acceptable carrier, excipient, or diluent.

[0258] In another aspect, provided herein is a method of treating a condition or disease in a subject in need therefor comprising administering to the subject a therapeutically effective amount of the multifunctional molecule as provided herein, the polynucleotide as provided herein, the vector as provided herein, the cell as provided herein, the pharmaceutical composition as provided herein, or a combination thereof, wherein the administering is effective to treat the condition or disease in the subject.

[0259] In some embodiments, the condition or disease is cancer. In some embodiments, the cancer is a solid tumor, a hematological cancer, a metastatic cancer, a soft tissue tumor, or a combination thereof.

[0260] In some embodiments, the cancer is the solid tumor, and wherein the solid tumor is selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

[0261] In some embodiments, the cancer is the hematological cancer, and wherein the hematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T- cell lymphoma, acute lymphocytic leukemia, and a combination thereof.WSGR Docket No.53676-769.601

[0262] In some embodiments, the Non-Hodgkin’s lymphoma is selected from the group consisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

[0263] In some embodiments, the T-cell lymphoma is peripheral T-cell lymphoma.

[0264] In some embodiments, the cancer is characterized by a cancer antigen present on the cancer.

[0265] In some embodiments, the cancer antigen is a tumor antigen, a stromal antigen, or a hematological antigen.

[0266] In some embodiments, the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, TRP-1 / -2, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, MUC-1, CA- 125, BAGE, GAGE, NY-ESO-1, β-catenin, CDK4, CDC27, α actinin-4, TRP1 / gp75, TRP2, gp100, Melan-A / MART1, gangliosides, WT1, EphA3, Epidermal growth factor receptor (EGFR), MART-2, MART-1, MUC1, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TRP1, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, delta-like ligand 3 (DLL3), CLDN18.2, and tenascin W.

[0267] In some embodiments, the cancer antigen is DLL3. In some embodiments, the cancer antigen is CLDN18.2.

[0268] In some embodiments, the method as provided herein further comprises administering a second therapeutic agent or therapy to the subject. In some embodiments, the second therapeutic agent or therapy comprises a chemotherapeutic agent, a biologic agent, a hormonal therapy, radiation, or surgery.

[0269] In some embodiments, the second therapeutic agent or therapy is administered in combination with the multifunctional molecule as provided herein, the polynucleotide as provided herein, the vector as provided herein, the cell as provided herein, or the pharmaceutical composition as provided herein, sequentially, simultaneously, or concurrently.

[0270] In another aspect, provided herein is the use of the multifunctional molecule as provided herein, the polynucleotide as provided herein, the vector as provided herein, the cell as provided herein, or the pharmaceutical composition as provided herein for the method as provided herein.WSGR Docket No.53676-769.601 INCORPORATION BY REFERENCE

[0271] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0272] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:

[0273] FIGS.1A-1V depict exemplary embodiments of multifunctional molecules as described herein.

[0274] FIGS.1A-1C depict exemplary embodiments of multifunctional molecules containing multiple, e.g., two, molecules of a cytokine molecule linked to an antibody molecule that binds to a T cell receptor beta variable region (TCRβV) (“anti-TCRβV antibody molecule”) or an antibody molecule that binds to a T cell receptor alpha variable region (TCRαV) (“anti-TCRαV antibody molecule”). FIG.1A depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to C- terminus of a VL of the first anti-TCRαV or anti-TCRβV Fab. The second cytokine molecule is fused to C-terminus of a VL of the second anti-TCRαV or anti-TCRβV Fab. FIG.1B depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to N-terminus of a VH of the first anti-TCRαV or anti-TCRβV Fab. The second cytokine molecule is fused to N-terminus of a VH of the second anti-TCRαV or anti-TCRβV Fab. FIG.1C depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to N-terminus of a VL of the first anti- TCRαV or anti-TCRβV Fab. The second cytokine molecule is fused to N-terminus of a VL of the second anti-TCRαV or anti-TCRβV Fab.

[0275] FIGS.1D-1F depict exemplary embodiments of multifunctional molecules containing a single molecule of a cytokine molecule linked to an anti-TCRβV antibody molecule or an anti-TCRαV antibody molecule. FIG.1D depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine molecule is fused to C-terminus of a VL of the first anti-TCRαV or anti-TCRβV Fab. FIG.1E depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine molecule is fused to N-terminus of a VH of the first anti-TCRαV or anti-TCRβV Fab. FIG.1FWSGR Docket No.53676-769.601 depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti- TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine molecule is fused to N-terminus of a VL of the first anti- TCRαV or anti-TCRβV Fab.

[0276] FIGS.1G-1J depict exemplary embodiments of multifunctional molecules containing a cytokine molecule linked to a first dimerization module. FIG.1G depicts an exemplary embodiment of a multifunctional molecule comprising an anti-TCRαV or anti-TCRβV 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-TCRαV or anti-TCRβV Fab 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.1H depicts an exemplary embodiment of a multifunctional molecule comprising an anti-TCRαV or anti-TCRβV 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-TCRαV or anti-TCRβV Fab is fused to the N-terminus of one Fc chain. The cytokine molecule is fused to the C-terminus of the other Fc chain. FIG.1I depicts an exemplary embodiment of a multifunctional molecule comprising a first anti- TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine molecule is fused to the C-terminus of one Fc chain. FIG.1J depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to the C-terminus of one Fc chain. The second cytokine molecule is fused to the C-terminus of the other Fc chain.

[0277] FIGS.1K-1M depict exemplary embodiments of multifunctional molecules containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation, and multiple, e.g., two, molecules of a cytokine molecule linked to an anti-TCRβV antibody molecule or an anti-TCRαV antibody molecule. FIG. 1K depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine is fused to the C-terminus of a VL of the first anti-TCRαV or anti-TCRβV Fab. The second cytokine is fused to the C-terminus of a VL of the second anti-TCRαV or anti-TCRβV Fab. FIG.1L depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti- TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine is fused to the N- terminus of a VH of the first anti-TCRαV or anti-TCRβV Fab. The second cytokine is fused to the N- terminus of a VH of the second anti-TCRαV or anti-TCRβV Fab. FIG.1M depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine is fused to the N-terminus of a VL of the firstWSGR Docket No.53676-769.601 anti-TCRαV or anti-TCRβV Fab. The second cytokine is fused to the N-terminus of a VL of the second anti-TCRαV or anti-TCRβV Fab.

[0278] FIGS.1N-1P depict exemplary embodiments of multifunctional molecules containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation (Knob-in-hole), and a single molecule of a cytokine molecule linked to an anti-TCRβV antibody molecule or an anti-TCRαV antibody molecule. FIG. 1N depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine is fused to the C-terminus of a VL of the first anti-TCRαV or anti-TCRβV Fab. FIG.1O depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine is fused to the N-terminus of a VH of the first anti-TCRαV or anti-TCRβV Fab. FIG.1P depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine is fused to the N-terminus of a VL of the first anti-TCRαV or anti-TCRβV Fab.

[0279] FIGS.1Q-1T depict exemplary embodiments of multifunctional molecules containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation (Knob-in-hole), and a cytokine molecule linked to the exemplary dimerization module. FIG. 1Q depicts an exemplary embodiment of a multifunctional molecule comprising an anti-TCRαV or anti-TCRβV 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-TCRαV or anti-TCRβV Fab 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.1R depicts an exemplary embodiment of a multifunctional molecule comprising an anti-TCRαV or anti-TCRβV 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-TCRαV or anti-TCRβV Fab is fused to the N-terminus of one Fc chain. The cytokine molecule is fused to the C- terminus of the other Fc chain. FIG.1S depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The cytokine molecule is fused to the C-terminus of one Fc chain. FIG.1T depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to the C-terminus of one Fc chain. The second cytokine molecule is fused to the C-terminus of the other Fc chain.

[0280] FIGS.1U-1V depict exemplary embodiments of multifunctional molecules containing an exemplary dimerization module, e.g., an Fc region comprising a N297A mutation (Knob-in-hole), and multiple, e.g., two, molecules of a cytokine molecule linked to an anti-TCRβV antibody molecule or an anti-TCRαV antibody molecule. FIG.1U depicts an exemplary embodiment of a multifunctionalWSGR Docket No.53676-769.601 molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to the C-terminus of one Fc chain. The second cytokine molecule is fused to the C-terminus of the other Fc chain. FIG.1V depicts an exemplary embodiment of a multifunctional molecule comprising a first anti-TCRαV or anti-TCRβV Fab, a second anti-TCRαV or anti-TCRβV Fab, a first cytokine molecule, a second cytokine molecule, and an Fc dimer. The Fc dimer comprises two Fc chains. The first cytokine molecule is fused to the C-terminus of a VL of the first anti- TCRαV or anti-TCRβV Fab. The second cytokine molecule is fused to the C-terminus of a VL of the second anti-TCRαV or anti-TCRβV Fab.

[0281] FIGS.2A and 2B shows the alignment of the Antibody A source mouse VH and VL framework 1, CDR 1, framework 2, CDR 2, framework 3, CDR3, and framework 4 regions with their respective humanized sequences. Kabat CDRs are shown in bold, Chothia CDRs are shown in italics, and combined CDRs are shown in boxes. The framework positions that were back mutated are double underlined. FIG. 2A shows VH sequences for murine Antibody A (SEQ ID NO: 1) and humanized Antibody A-H (SEQ ID NO: 9). FIG.2B shows VL sequences for murine Antibody A (SEQ ID NO: 2) and humanized Anti- body A-H (SEQ ID NO: 10 and SEQ ID NO: 11).

[0282] FIGS.2C-2D shows the alignment of the Antibody B source mouse VH and VL framework 1, CDR 1, framework 2, CDR 2, framework 3, CDR3, and framework 4 regions with their respective humanized sequences. Kabat CDRs are shown in bold, Chothia CDRs are shown in italics, and combined CDRs are shown in boxes. The framework positions that were back mutated are double underlined. FIG. 2C shows the VH sequence for murine Antibody B (SEQ ID NO: 15) and humanized VH sequences B- H.1A to B-H.1C (SEQ ID NOs: 23-25). FIG.2D shows the VL sequence for murine Antibody B (SEQ ID NO: 16) and humanized VL sequences B-H.1D to B-H.1H (SEQ ID NOs: 26-30).

[0283] FIG.3 shows alignment of TCRBV amino acid sequences (SEQ ID NOS 3557-3739, respectively, in order of appearance). The alignment of TCRBV amino acid sequences underscores the diversity of TCR sequences. In particular, the TRBV sequences from different subfamilies are considerably different from each other.

[0284] FIG.4 depicts the phylogenetic tree of TCRBV gene family and subfamilies with corresponding antibodies mapped. Subfamily identities are as follows: Subfamily A: TCRβ V6; Subfamily B: TCRβ V10; Subfamily C: TCRβ V12; Subfamily D: TCRβ V5; Subfamily E: TCRβ V7; Subfamily F: TCRβ V11; Subfamily G: TCRβ V14; Subfamily H: TCRβ V16; Subfamily I:TCRβ V18; Subfamily J:TCRβ V9; Subfamily K: TCRβ V13; Subfamily L: TCRβ V4; Subfamily M:TCRβ V3; Subfamily N:TCRβ V2; Subfamily O:TCRβ V15; Subfamily P: TCRβ V30; Subfamily Q: TCRβ V19; Subfamily R:TCRβ V27; Subfamily S:TCRβ V28; Subfamily T: TCRβ V24; Subfamily U: TCRβ V20; Subfamily V: TCRβ V25; and Subfamily W:TCRβ V29 subfamily. Subfamily members are described in detail herein in the Section titled “TCR beta V (TCRβV)”.

[0285] FIGS.5A-5C show human CD3+ T cells activated by anti-TCR Vβ13.1 antibody (A-H.1) for 6- days. Human CD3+ T cells were isolated using magnetic-bead separation (negative selection) andWSGR Docket No.53676-769.601 activated with immobilized (plate-coated) anti-TCR Vβ13.1 (A-H.1) or anti-CD3^ (OKT3) antibodies at 100 nM for 6 days. FIG.5A shows two scatter plots (left: activated with OKT3; and right: activated with A-H.1) of expanded T cells assessed for TCR Vβ13.1 surface expression using anti-TCR Vβ13.1 (A-H.1) followed by a secondary fluorochrome- conjugated antibody for flow cytometry analysis. FIG.5B shows percentage (%) of TCR Vβ13.1 positive T cells activated by anti-TCR Vβ13.1 (A-H.1) or anti-CD3e (OKT3) plotted against total T cells (CD3+). FIG.5C shows relative cell count acquired by counting the number of events in each T cell subset gate (CD3 or TCR Vβ13.1) for 20 seconds at a constant rate of 60μl / min. Data shown as mean value from 3 donors.

[0286] FIGS.6A-6B show cytolytic activity of human CD3+ T cells activated by anti-TCR Vβ13.1 antibody (A-H.1) against transformed cell line RPMI 8226. FIG.6A depicts target cell lysis of human CD3+ T cells activated with A-H.1or OKT3. Human CD3+ T cells were isolated using magnetic-bead separation (negative selection) and activated with immobilized (plate-coated) A-H.1 or OKT3 at the indicated concentrations for 4 days prior to co-culture with RPMI 8226 cells at a (E:T) ratio of 5:1 for 2 days. Samples were next analyzed for cell lysis of RPMI 8226 cells by FACS staining for CFSE / CD138- labeled, and membrane-impermeable DNA dyes (DRAQ7) using flow cytometry analysis. FIG.6B shows target cell lysis of human CD3+ T cells activated with A-H.1 or OKT3 incubated with RPMI-8226 at a (E:T) ratio of 5:1 for 6 days followed by cell lysis analysis of RPMI 8226 cells as described above. Percentage (%) target cell lysis was determined by normalizing to basal target cell lysis (i.e., without antibody treatment) using the following formula, [(x - basal) / (100% - basal), where x is cell lysis of sample]. Data shown is a representative of n=1 donor.

[0287] FIGS.7A-7B show IFNγ production by human PBMCs activated with the indicated antibodies. Human PBMCs were isolated from whole blood from the indicated number of donors, followed by solid- phase (plate-coated) stimulation with the indicated antibodies at 100Nm. Supernatant was collected on Days 1, 2, 3, 5, or 6. FIG.7A is a graph comparing the production of IFNγ in human PBMCs activated with the antibodies indicated activated with anti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) or anti-CD3e antibodies (OKT3 or SP34-2) on Day 1, 2, 3, 5, or 6 post-activation. FIG.7B shows IFNγ production in human PBMCs activated with the antibodies indicated activated with the indicated anti-TCR Vβ13.1 antibodies or anti-CD3e antibody (OKT3) on Day 1, 2, 3, 5, or 6 post-activation.

[0288] FIGS.8A-8B show IL-2 production by human PBMCs activated with the indicated antibodies. A similar experimental setup as described for FIG.8A is a graph comparing the production of IL-2 in human PBMCs activated with the antibodies indicated activated with anti-TCR Vβ13.1 antibodies (A- H.1 or A-H.2) or anti-CD3e antibodies (OKT3 or SP34-2) on Day 1, 2, 3, 5, or 6 post-activation. FIG. 8B shows IL-2 production in human PBMCs activated with the antibodies indicated activated with the indicated anti-TCR Vβ13.1 antibodies or anti-CD3e antibody (OKT3) on Day 1, 2, 3, 5, or 6 post- activation.

[0289] FIGS.9A- 9B show IL-6 production by human PBMCs activated with the indicated antibodies. FIG.9A is a graph comparing the production of IL-6 in human PBMCs activated with the antibodies indicated activated with anti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) or anti-CD3e antibodies (OKT3WSGR Docket No.53676-769.601 or SP34-2) on Day 1, 2, 3, 5, or 6 post-activation. FIG.9B shows IL-6 production in human PBMCs activated with the antibodies indicated activated with the indicated anti-TCR Vβ13.1 antibodies or anti- CD3e antibody (OKT3) on Day 1, 2, 3, 5, or 6 post-activation.

[0290] FIGS.10A- 10B show TNF-alpha production by human PBMCs activated with the indicated antibodies. FIG.10A is a graph comparing the production of TNF-alpha in human PBMCs activated with the antibodies indicated activated with anti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) or anti-CD3e antibodies (OKT3 or SP34-2) on Day 1, 2, 3, 5, or 6 post-activation. FIG.10B shows TNF-alpha production in human PBMCs activated with the antibodies indicated activated with the indicated anti- TCR Vβ13.1 antibodies or anti-CD3e antibody (OKT3) on Day 1, 2, 3, 5, or 6 post-activation.

[0291] FIGS.11A- 11B show IL-1beta production by human PBMCs activated with the indicated antibodies. FIG.11A is a graph comparing the production of IL-1beta in human PBMCs activated with the antibodies indicated activated with anti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) or anti-CD3e antibodies (OKT3 or SP34-2) on Day 1, 2, 3, 5, or 6 post-activation. FIG.11B shows IL-1beta production in human PBMCs activated with the antibodies indicated activated with the indicated anti- TCR Vβ13.1 antibodies or anti-CD3e antibody (OKT3) on Day 1, 2, 3, 5, or 6 post-activation.

[0292] FIGS.12A-12B are graphs showing delayed kinetics of IFNγ secretion in human PMBCs activated by anti-TCR Vβ13.1 antibody A-H.1 when compared to PBMCs activated by anti-CD3e antibody OKT3. FIG.12A shows IFNγ secretion data from 4 donors. FIG.12B shows IFNγ secretion data from 4 additional donors. Data shown is representative of n=8 donors.

[0293] FIG.13 depicts increased CD8+ TSCM and Temra T cell subsets in human PBMCs activated by anti-TCR Vβ13.1 antibodies (A-H.1 or A-H.2) compared to PBMCs activated by anti-CD3e antibodies (OKT3 or SP34-2).

[0294] FIGS.14A-14F show characterization of an anti-TCRVb antibody. FIG.14A is a graph depicting proliferation of T cells activated with anti-CD3 (OKT3) antibody or anti-TCRVb antibody. FIG. 14B shows selective expansion of CD45RA+ effector memory CD8+ and CD4+ T cells (TEMRA) cells with anti- TCRVb antibodies. Tn= naïve T cell; Tscm= stem cell memory T cell; Tcm= central memory T cell; Tem=effector memory T cell; Temra=effector memory CD45RA+ T cell. FIG.14C is a graph showing IFN-g secretion by PBMCs stimulated with an anti-TCRVb antibody, or anti-CD3 antibodies. FIG.14D shows target cell lysis by T cells stimulated with an anti-TCRVb antibody, or anti-CD3 antibodies. Cells were stimulated for 4 days followed by 2 days incubation with multiple myeloma target cells for assessment of cell killing. FIG.14E is a graph showing perforin secretion by T cells stimulated with an anti-TCRVb antibody, or an anti-CD3 antibody. Perforin was analyzed by FACS staining in TCRVB-positive and TCRVB-negative T cells in PBMCs after 5 days of stimulation with 100ng / ml plate-bound antibody. FIG.14F is a graph showing Granzyme B by T cells stimulated with an anti- TCRVb antibody, or an anti-CD3 antibody. Granzyme B was analyzed by FACS staining in TCRVB- positive and TCRVB-negative T cells in PBMCs after 5 days of stimulation with 100ng / ml plate-bound antibody.WSGR Docket No.53676-769.601

[0295] FIGS.15A-15B show production of IL-2 and IL-15 and expansion of human NK cells by stimulation of PBMCs with anti-TCRVb antibody for 6 days at a dose of 100nM. FIG.15A shows secretion of IL-2 or IL-15 in T cells stimulated with an anti-TCRVb antibody, or anti-CD3 antibodies. FIG.15B depicts flow cytometry dot plots showing NKp46 staining vs CD56 antibody staining in cells stimulated with an anti-TCRVb antibody or an anti-CD3 antibody or a control sample.

[0296] FIGS.16A-16C show secretion of cytokines in PBMCs stimulated with an anti-TCRVb antibody, or anti-CD3 antibodies.

[0297] FIGS.17A-17B show killing of MM cells by dual targeting BCMA-TCRvb antibody molecules. FIG.17A shows in vitro killing by one of the following dual-targeting antibody molecules: BCMA- TCRVb (Molecule I), BCMA-CD3, or Control-TCRVb; or an isotype control. FIG.17B shows in vivo killing of MM cells by a dual-targeting BCM-TCRVb antibody (Molecule I).

[0298] FIG.18 shows lysis of MM target cells with a dual targeting antibody (Molecule E) which recognized FcRH5 on one arm and TCRVb on the other arm.

[0299] FIGS.19A-19B demonstrate cytokine production from human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) when compared to those activated by anti-CD3^ antibodies (OKT3 or SP34-2). FIG.19A shows that human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) produce similar or reduced levels of IFN ^^. FIG.19B shows human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) produce higher levels of IL-2 when compared to those activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representative of n = 6 donors.

[0300] FIGS.20A-20C demonstrate cytokine production from human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1). Human PBMCs activated by anti-TCR Vβ8a antibodies (B-H.1) do not significantly produce IL-6 (FIG.20A), IL1β (FIG.20B), and less TNFα (FIG.20C), when compared to PBMCs activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representative of n = 6 donors.

[0301] FIGS.21A-21E demonstrate cytokine production from human PBMCs activated by anti-TCRβV Antibody D antibody compared to control anti-CD3e antibody (OKT3). FIG.21A shows that human PBMCs activated by anti-TCRβV Antibody D antibody produce similar or reduced levels of IFN ^^. FIG. 21B shows human PBMCs activated by anti-TCRβV Antibody D antibody produce higher levels of IL-2 when compared to those activated by anti-CD3^ antibodies (OKT3). Human PBMCs activated by anti- TCRβV Antibody D antibody do not significantly produce IL-1beta (FIG.21C), IL-6, (FIG.21D), or TNFalpha (FIG.21E). Data shown is representative of n = 4 donors.

[0302] FIGS.22A-22B demonstrate cytokine production from human PBMCs activated by anti-TCR Vβ5 antibody (Antibody E). FIG.22A shows that human PBMCs activated by anti-TCR Vβ5 antibody produce similar or reduced levels of IFN ^^ compared to PBMCS activated by anti-CD3^ antibodies (OKT3 or SP34-2). FIG.22B shows human PBMCs activated by the anti- TCR Vβ51 antibody produce higher levels of IL-2 when compared to those activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representative of n = 4 donors.WSGR Docket No.53676-769.601

[0303] FIGS.23A-23D demonstrate cytokine production from human PBMCs activated by an anti-TCR Vβ5 antibody (Antibody E). Human PBMCs activated by anti-TCR Vβ5 antibody do not significantly produce IL-1beta (FIG.23A), IL-6, (FIG.23B), TNFalpha (FIG.23C), or IL-10 (FIG.23D) as compared to PBMCs activated by anti-CD3^ antibodies (OKT3 or SP34-2). Data shown is representative of n = 4 donors.

[0304] FIGS.24A-24F demonstrate cytokine production from human PBMCs activated by a dual targeting (bispecific molecule) comprising an anti-TCRβV binding moiety and a BCMA binding moiety. FIG.24A shows that human PBMCs activated by the bispecific molecule produce similar or reduced levels of IFN ^^ as PBMCS activated by anti-CD3^ antibodies (OKT3). FIG.24B shows human PBMCs activated by the bispecific molecule produce higher levels of IL-2 when compared to PBMCs activated by anti-CD3^ antibodies (OKT3). Human PBMCs activated by the bispecific molecule do not significantly produce IL-1beta (FIG.24C), IL-6, (FIG.24D), TNFalpha (FIG.24E), or IL-10 (FIG. 24F). Data shown is representative of n = 3 donors.

[0305] FIGS.25A-25B show the structure and sequence of eight TCRβV proteins from seven different subfamilies: TCRβV6 subfamily (TCRβV6-5 and TCRβV6-4 are shown), TCRβV28 subfamily, TCRβV19 subfamily, TCRβV9 subfamily, TCRβV5 subfamily, TCRβV20 subfamily and TCRβV12 subfamily. FIG.25A shows the structural alignment of the different TCRβV proteins. The circled area represents the outward facing region comprising the proposed binding site for the anti-TCRβV antibodies as described herein. FIG.25B shows the amino acid sequence alignment of the proteins shown in FIG. 25A (SEQ ID NOS 3449-3456, respectively, in order of appearance). The various TCRβV proteins (from 7 different TCRβV subfamilies) have diverse sequences but share a conserved (similar) structure and function.

[0306] FIGS.26A-26J show secretion of IFNg (FIG.26A), IL-2 (FIG.26B), IL-1beta (FIG.26C), IL-6 (FIG.26D), IL-10 (FIG.26E), IL-4 (FIG.26F), TNFα (FIG.26G), IL-12p70 (FIG. 26H), IL-13 (FIG. 26I), IL-8 (FIG.26J) in PBMCs activated with anti-TCRVb antibodies (A-H.1, B-H.1), a bispecific molecule comprising an anti-TCRVb antibody (Molecule H), control isotype (122) or anti-CD3e antibody (OKT3). Data shown is representative of n = 2 donors and representative of 2 independent experiments.

[0307] FIGS.27A-27H show secretion of Eotaxin (FIG.27A), Eotaxin-3 (FIG.27B), IL-8(HA) (FIG. 27C), IP-10 (FIG.27D), MCP-1 (FIG.27E), MCP-4 (FIG.27F), MDC (FIG.27G), and MIP-1a (FIG. 27H) in PBMCs activated with anti-TCRVb antibodies (A-H.1, B-H.1), a bispecific molecule comprising an anti-TCRVb antibody (Molecule H), control isotype (122) or anti-CD3e antibody (OKT3). Data shown is representative of n = 2 donors and representative of 2 independent experiments.

[0308] FIGS.28A-28L show secretion of MIP-1B (FIG.28A), TARC (FIG.28B), GM-CSF (FIG. 28C), IL-12-23p40 (FIG.28D), IL-15 (FIG.28E), IL-16 (FIG.28F), IL-17a (FIG.28G), IL-1a (FIG. 28H), IL-5 (FIG.28I), IL-7 (FIG.28J), TNF-B (FIG.28K), and VEGF (FIG.28L) in PBMCs activated with anti-TCRVb antibodies (A-H.1, B-H.1), a bispecific molecule comprising an anti-TCRVb antibodyWSGR Docket No.53676-769.601 (Molecule H), control isotype (122) or anti-CD3e antibody (OKT3). Data shown is representative of n = 2 donors and representative of 2 independent experiments.

[0309] FIG.29 is a graph depicting mean tumor volume in NOD / SCID / IL-2Rγnull (NSG) mice engrafted with Raji-luc cells at days 10 to 28. The Star denotes PBMC implantation. Open triangles denote antibody treatment with the indicated antibodies.

[0310] FIGS.30A-30F are graphs showing cytokine secretion stimulated by anti-TRBC1 (Antibody F) or anti-CD3 (OKT3) at Days 2 and 5. Cytokines examined include: IFNγ (FIG.30A), IL-2 (FIG.30B), IL-1β (FIG.30C), IL-6 (FIG.30D), IL-10 (FIG.30E), and TNFα (FIG.31F).

[0311] FIG.31 is a FACS plot showing the expansion of TCRvb 6-5+ T cells over 8 days using anti- TCRvb 6-5 v1.

[0312] FIG.32 is a bar graph showing the expansion of TCRvb 6-5+ CD4+ T cells and TCRvb 6-5+ CD8+ T cells over 8 days using the anti-CD3ε antibody OKT3 (100nM).

[0313] FIG.33 is a bar graph showing the expansion of TCRvb 6-5+ CD4+ T cells and TCRvb 6-5+ CD8+ T cells over 8 days using the anti-TCRvb 6-5 v1 antibody (100nM).

[0314] FIG.34 is a FACS plot showing the showing the expansion of TCRvb 6-5+ T cells over 8 days using anti-TCRvb 6-5 v1 or the anti-CD3ε antibody OKT3.

[0315] FIG.35A is a bar graph showing the percentage of TCRβV 6-5+ T cells in PBMC cultures after 8 days of culture with the indicated antibody. Data for 5 replicates are shown. FIG.35B is a bar graph showing the percentage of TCRβV 6-5+ T cells in purified T cell cultures after 8 days of culture with the indicated antibody. Data for 5 replicates are shown.

[0316] FIG.36A is a bar graph showing the relative count of TCRβV 6-5+ T cells in PBMC culture after 8 days of culture with the indicated antibody. FIG.36B is a bar graph showing the relative count of TCRβV 6-5+ T cells in PBMC culture after 8 days of culture with the indicated antibody.

[0317] FIG.37A is a bar graph showing the relative count of TCRβV 6-5+ T cells in a purified T cell culture after 8 days of culture with the indicated antibody. FIG.37B is a bar graph showing the relative count of TCRβV 6-5+ T cells in a purified T cell culture after 8 days of culture with the indicated antibody.

[0318] FIG.38 is a line graph showing the total CD3+ T cell count (fold increase) after 8 days of T cell culture with either the anti-CD3ε antibody OKT3 or the anti-TCRvb 6-5 v1 antibody.

[0319] FIG.39 is a series of line graphs showing the kinetics of target cells by TCRβV 6-5 v1 activated T cells or anti-CD3ε (OKT3) activated T cells. T cells from three different donors were utilized (donor 6769, donor 9880, donor 5411).

[0320] FIG.40A is a scatter plot showing the percent of target cell lysis by T cells by TCRβV 6-5 v1 activated T cells or anti-CD3ε (OKT3) activated T cells without T cell pre activation. The data is presented at day 6 of co-culture between target cells and effector T cells. FIG.40B is a scatter plot showing the percent of target cell lysis by T cells by TCRβV 6-5 v1 activated T cells or anti-CD3ε (OKT3) activated T cells with 4 days of T cell pre activation. The data is presented at day 2 of co-culture between target cells and effector T cells (after 4 days of T cell pre-activation).WSGR Docket No.53676-769.601

[0321] FIG.41 is a scatter plot showing the percent of target cell lysis by T cells by TCRβV 6-5 v1 activated T cells or anti-CD3ε (OKT3) activated T cells with 4 days of T cell pre activation. The data is presented at day 2 of co-culture between target cells and effector T cells (after 4 days of T cell pre- activation).

[0322] FIG.42 is a bar graph showing target cell lysis by T cells by TCRβV 6-5 v1 activated T cells or anti-CD3ε (OKT3) activated T cells (100nM each antibody). The data includes seven replicates of each experimental condition.

[0323] FIG.43 is a series of FACS plots that show the cell surface expression of CD3ε on CD4+ TCRβV 6-5- or CD4+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0324] FIG.44 is a series of FACS plots that show the cell surface expression of CD3ε on CD8+ TCRβV 6-5- or CD8+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0325] FIG.45 is a series of FACS plots that show the cell surface expression of TCRβV on CD4+ TCRβV 6-5- or CD4+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0326] FIG.46 is a series of FACS plots that show the cell surface expression of TCRβV on CD8+ TCRβV 6-5- or CD8+ TCRβV 6-5+T cells activated with either SP34-2 (anti-CD3ε antibody) or anti- TCRβV 6-5 v1 (anti- TCRβV 6-5 antibody) at days 0, 1, 2, 4, 6, or 8 post antibody activation.

[0327] FIG.47A shows FACS plot of TCRβV 6-5+cynomolgus T cell expansion either unstimulated (left) or stimulated with anti-TCRβV 6-5 v1 (right) 7 days post activation of cynomolgus PBMCs. PBMCs from Donor DW8N (fresh PBMC sample, male, age 8, weight 7.9 kgs) were used. FIG.47B shows FACS plot of TCRβV 6-5+cynomolgus T cell expansion either unstimulated (left) or stimulated with anti-TCRβV 6-5 v1 (right) 7 days post activation of cynomolgus PBMCs. PBMCs from Donor G709 (cryopreserved sample, male, age 6, weight 4.7 kgs) were used.

[0328] FIG.48 shows FACS plot and corresponding microscopy images of TCRβV 6-5+cynomolgus T cell expansion either unstimulated (left), stimulated with SP34-2 (anti-CD3ε antibody) (middle); or stimulated with anti-TCRβV 6-5 v1 (right) post activation of cryopreserved donor DW8N cynomolgus PBMCs. The microscopy images show the cell cluster formation (indicated by circles).

[0329] FIG.49 shows a schematic of FACS plot showing the FACS gating / staining of PBMCs prior ɣ^ T cell purification.

[0330] FIG.50 shows a schematic of FACS plot showing the FACS gating / staining of purified ɣ^ T cell population.

[0331] FIG.51 show activation of purified ɣ^ T cell population with anti-CD3ε antibody (SP34-2) (left) or anti-TCRβV antibody (anti-TCRβV 6-5 v1) (right).

[0332] FIG.52A shows the release of IFNɣ from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52B shows the release of TNFα from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-WSGR Docket No.53676-769.601 TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52C shows the release of IL-2 from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti- TCRβV 6-5 v1), or unstimulated. FIG.52D shows the release of IL-17A from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52E shows the release of IL-1α from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52F shows the release of IL-1β from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34- 2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52G shows the release of IL-6 from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated. FIG.52H shows the release of IL-10 from purified ɣ^ T cell populations activated with anti-CD3ε antibody (SP34-2), anti-TCRβV antibody (anti-TCRβV 6-5 v1), or unstimulated.

[0333] FIG.53 shows the relative representations of all TCR alpha V segments (TRAV group of genes) and their variants (top), all TCR beta V segment 6-5 variants (TRBV6-5 gene) (bottom left), and all TCR beta V segments and variants excluding 6-5 (bottom right).

[0334] FIG.54A is a FACS plot showing phenotypic markers of CD4+ T cells expanded with anti- TCRβV antibody (anti-TCRβV 6-5 v1). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right). FIG.54B is a FACS plot showing phenotypic markers of CD4+ T cells expanded with anti-CD3ε antibody (OKT3). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right).

[0335] FIG.55A is a FACS plot showing phenotypic markers of CD8+ T cells expanded with anti- TCRβV antibody (anti-TCRβV 6-5 v1). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right). FIG.55B is a FACS plot showing phenotypic markers of CD8+ T cells expanded with anti-CD3ε antibody (OKT3). Defined phenotypes include TEMRA (top left), Naïve / TSCM (top right), TEM (bottom left), and TCM (bottom right).

[0336] FIG.56A is a bar graph showing the percentage of PD1 expressing CD4+ T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated. FIG.56B is a bar graph showing the percentage of PD1 expressing CD8+ T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0337] FIG.57A is a bar graph showing the expression of Ki-67 by CD4+ T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated. FIG.57B is a bar graph showing the expression of Ki-67 by CD8+ T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0338] FIG.58A is a FACS plot showing the percentage of TEMRA-like CD8+ T cells activated using anti-TCRβV antibody (anti-TCRβV 6-5 v1) that express CD57 (18.7%). FIG.58B is a FACS plot showing the percentage of TEM-like CD8+ T cells activated using anti-CD3ε antibody (OKT3) thatWSGR Docket No.53676-769.601 express CD57 (46.8%) and the percentage of TCM-like CD8+ T cells activated using anti-CD3ε antibody (OKT3) that express CD57 (18.9%).

[0339] FIG.59 shows a series of FACS plots showing the expression of expression of CD27 and by CD4+ (top) or CD8+ (bottom) T cells from T cell cultures activated with anti-TCRβV antibody (anti- TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0340] FIG.60 shows a series of FACS plots showing the expression of expression of OX40, 41BB, and ICOS by CD4+ (top) or CD8+ (bottom) T cells from T cell cultures activated with anti-TCRβV antibody (anti-TCRβV 6-5 v1), anti-CD3ε antibody (OKT3), or unstimulated.

[0341] FIG.61 shows a series of FACS plots showing the percentage of CD3+ (CD4 gated) TCRβV 6- 5+ T cells 1, 2, 3, 4, 5, 6, and 8 days port activation with BCMA and the anti-TCR Vβ antibody anti-TCR Vβ 6-5 v1.

[0342] FIG.62A shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 0 post activation. FIG.62B shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 1 post activation. FIG.62C shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 2 post activation. FIG.62D shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti- TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 3 post activation. FIG.62E shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 4 post activation. FIG.62F shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 5 post activation. FIG.62G shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti- CD3ε (OKT3) antibodies on day 6 post activation. FIG.62H shows a series of FACS plots showing the percentage of CD4+ T cells expanded using isotype control (IgG1 N297A), anti-TCRβV (anti-TCR Vβ 6-5 v1), or anti-CD3ε (OKT3) antibodies on day 8 post activation.

[0343] FIG.63A is a bar graph showing ATP production from glycolysis of T cell cultures activated with the indicated antibodies. FIG.63B is a bar graph showing ATP production from oxidative phosphorylation of T cell cultures activated with the indicated antibodies.

[0344] FIG.64 is a line graph showing the oxygen consumption rate (OCR) of T cells from about 0 to 75 minutes activated with the indicated antibody.

[0345] FIG.65A shows the oxygen consumption rate (OCR) of T cells activated with the indicated antibody during basal respiration. FIG.65B shows the oxygen consumption rate (OCR) of T cells activated with the indicated antibody during maximal respiration. FIG.65C shows the oxygen consumption rate (OCR) of T cells activated with the indicated antibody during spare respiratoryWSGR Docket No.53676-769.601 capacity. FIG.65D is a line graph indicates the areas of basal respiration and maximal respiration as shown in FIG.64A and FIG.64B, respectively.

[0346] FIG.66A is a bar graph showing ATP production from glycolysis of T cell cultures activated with anti-TCRβV 6-5 v1 and re-stimulated with the indicated antibody. FIG.66B is a bar graph showing ATP production from oxidative phosphorylation of T cell cultures activated with anti-TCRβV 6-5 v1 and re-stimulated with the indicated antibody.

[0347] FIGS.67A-67G are graphs showing expression of IFNγ (FIG.67A), TNFα (FIG.67E), IL-1α (FIG.67B), IL-1β (FIG.67C), IL-6 (CRS and neurotoxicity associated cytokines) (FIG.67D) with BHM1710 (anti TCRVB), a reduced affinity anti CD3 antibody (TB) and the SP34 anti CD3e antibody. IL-10 (FIG.67F), IL-17A (FIG.67G).

[0348] FIG.68 is a FACS plot showing the percentage of NK cells expanded from T cell cultures activated with the indicated antibody.

[0349] FIG.69 is a bar graph showing the number of NK cells expanded from T cell cultures activated with the indicated antibody.

[0350] FIG.70 shows a series of FACS plots showing NK cell proliferation induced by T cell cultures activated with the indicated antibody.

[0351] FIG.71 is a schematic showing an assay described in Example for determining NK cell mediated lysis of target K562 cells.

[0352] FIG.72 is a bar graph showing the percent target cell lysis mediated by NK cells activated by PBMCs activated with the indicated antibody.

[0353] FIG.73 shows a series of FACS plots showing the proliferation of NK cells from PBMC cultures activated / expanded with the indicated antibody (isotype control or OKT3). PBMCs from three donors (D1, D2, and D3) were analyzed.

[0354] FIG.74 shows a series of FACS plots showing the proliferation of NK cells from PBMC cultures activated / expanded with the indicated antibody (anti-TCRv ^ 12-3 / 4 v1 or anti-TCRv ^ 12-3 / 4 v2). PBMCs from three donors (D1, D2, and D3) were analyzed.

[0355] FIG.75 shows a series of FACS plots showing the proliferation of NK cells from PBMC cultures activated / expanded with the indicated antibody (anti-TCRv ^ 12-3 / 4 v3 or SP34-2). PBMCs from three donors (D1, D2, and D3) were analyzed.

[0356] FIG.76 is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0357] FIG.77 is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0358] FIG.78 is a bar graph showing the level of secreted IL-15 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).WSGR Docket No.53676-769.601

[0359] FIG.79 is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0360] FIG.80 is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0361] FIG.81 is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34) and cultured with said antibody for the indicated number of days (1, 3, or 5).

[0362] FIG.82 is a bar graph showing the level of the indicated cytokine secreted by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or SP34). The data includes use of 17 individual PBMC donors.

[0363] FIG.83A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83C is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.83G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or OKT3) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6).

[0364] FIG.84A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84C is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control)WSGR Docket No.53676-769.601 and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti- TCRβV 6-5 v1, OKT3, SP34-2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6). FIG.84G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, SP34- 2, or isotype control) and cultured with said antibody for the indicated number of days (1, 2, 3, 5, or 6).

[0365] FIG.85A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85C is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG. 85D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti- TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.85F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG. 85G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8).

[0366] FIG.86A is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (2, 5, or 7). FIG.86B is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (2, 5, or 8). FIG.86C is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1, OKT3, or SP34-2) and cultured with said antibody for the indicated number of days (2, 5, or 7). FIG.86D is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 or SP34-2) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7).WSGR Docket No.53676-769.601

[0367] FIG.87A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87B is a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87C is a bar graph showing the level of secreted IL- 4 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87F is a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87G is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87H is a bar graph showing the level of secreted IL-12p70 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87I is a bar graph showing the level of secreted IL- 13 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87J is a bar graph showing the level of secreted IL-8 by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87K is a bar graph showing the level of secreted exotaxin by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87L is a bar graph showing the level of secreted exotoxin-3 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2,WSGR Docket No.53676-769.601 3, 4, 5, 6, or 8). FIG.87M is a bar graph showing the level of secreted IL-8 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87N is a bar graph showing the level of secreted IP- 10 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87O is a bar graph showing the level of secreted MCP-1 by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87P is a bar graph showing the level of secreted MCP-4 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87Q is a bar graph showing the level of secreted MDC by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87R is a bar graph showing the level of secreted MIP-1a by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87S is a bar graph showing the level of secreted MIP-1b by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87T is a bar graph showing the level of secreted TARC by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87U is a bar graph showing the level of secreted GMCSF by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87V is a bar graph showing the level of secreted IL-12-23p40 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87W is a bar graph showing the level of secreted IL-15 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87X is a bar graph showing the level of secreted IL- 16 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with saidWSGR Docket No.53676-769.601 antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87Y is a bar graph showing the level of secreted IL-17a by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87Z is a bar graph showing the level of secreted IL-1a by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87AA is a bar graph showing the level of secreted IL-5 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87BB is a bar graph showing the level of secreted IL-7 by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87CC is a bar graph showing the level of secreted TNF-B by T cells activated / expanded with the indicated antibody (isotype control; anti-TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8). FIG.87DD is a bar graph showing the level of secreted VEGF by T cells activated / expanded with the indicated antibody (isotype control; anti- TCRβV 6-5 v1 with anti-BCMA antibody; anti-TCRβV 6-5 v1; anti-TCRβV 123 / 4 v1, or SP34-2) and cultured with said antibody for the indicated number of days (1, 2, 3, 4, 5, 6, or 8).

[0368] FIG.88 shows a graphical representation of the relation of sequences between different TCRVB clonotype subfamilies.

[0369] FIG.89A is a bar graph showing the percentage of cytokine release from PBMCs activated / expanded for eight days using the indicated antibody (anti-TCRβV 12-3 / 4 v1 or SP34-2). FIG. 89B is a bar graph showing the percentage of cytokine release from PBMCs activated / expanded for eight days using the indicated antibody (anti-TCRβV 5 or SP34-2). FIG.89C is a bar graph showing the percentage of cytokine release from PBMCs activated / expanded for eight days using the indicated antibody (anti-TCRβV 10 or SP34-2).

[0370] FIG.90A a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90B a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90C a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90D a bar graph showing the level of secreted IL-1α by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90E a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90F a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90G a bar graph showing the level of secreted TNFα by TWSGR Docket No.53676-769.601 cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.90H a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6).

[0371] FIG.91 is a bar graph summarizing data from FACS analysis of PBMCs activated / expanded for 6 days using the indicated anti-TCRV ^ antibody.

[0372] FIG.92A a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92B a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92C a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92D a bar graph showing the level of secreted IL-1α by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92E a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92F a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92G a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7). FIG.92H a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody for the indicated number of days (1, 3, 5, or 7).

[0373] FIG.93 is a bar graph summarizing data from FACS analysis of PBMCs activated / expanded for 7 days using the indicated anti-TCRV ^ antibody.

[0374] FIG.94A is a bar graph showing the level of secreted IFNɣ by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94B a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94C a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94D a bar graph showing the level of secreted IL-1α by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94E a bar graph showing the level of secreted IL-1β by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94F a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94G a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94H a bar graph showing the level of secreted TNFα by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6). FIG.94I a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody for the indicated number of days (3 or 6).

[0375] FIG.95A is a bar graph showing the level of secreted IFN-ɣ by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95B is a bar graph showing the level of secreted IFN-ɣ by T cells activated / expandedWSGR Docket No.53676-769.601 with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6- 5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95C is a bar graph showing the level of secreted IL-1b by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95D is a bar graph showing the level of secreted IL-6 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95E is a bar graph showing the level of secreted IL-10 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95F is a bar graph showing the level of secreted IL-15 by T cells activated / expanded with the indicated antibody (anti- TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95G is a bar graph showing the level of secreted IL-17A by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95H is a bar graph showing the level of secreted IL-1a by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95I is a bar graph showing the level of secreted IL-1b by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6- 5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95J is a bar graph showing the level of secreted IL-2 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95K is a bar graph showing the level of secreted IL-4 by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7). FIG.95L is a bar graph showing the level of secreted TNF-a by T cells activated / expanded with the indicated antibody (anti-TCRβV 6-5 v1 (plate coated), anti-CD3ε (plate coated), anti-TCRβV 6-5 v1 (in solution), or anti-CD3ε (in solution) and cultured with said antibody for the indicated number of days (1, 3, 5, or 7).

[0376] FIG.96 is a FACS plot showing the showing the ability of MH3-2 to bind PBMCs from one of two donors when the PBMCs are either preincubated with TM23 or not (MH3-2 Alone).

[0377] FIG.97 is a FACS plot showing the ability of MH3-2 to bind PBMCs from one of two donors when the PBMCs are either preincubated with TM23 or not (MH3-2 Alone).WSGR Docket No.53676-769.601

[0378] FIG.98A is a bar graph showing the polyfunctional strength index (PSI) of PBMC CD4+ T cells, CD4+ T cells expanded with anti-CD3 antibody, (CD3 Expanded T cells), and CD4+ T cells expanded with anti-TCRVβ 6-5 antibody (Drug Expanded T cells). The Effector mediators are Granzyme B, IFNɣ, MIP-1α, perforin, TNFα, and TNFβ. The Stimulatory mediators are IL-5. The Chemoattractive mediators are MIP-1b. FIG.98B is a bar graph showing the polyfunctional strength index (PSI) of PBMC CD8+ T cells, CD8+ T cells expanded with anti-CD3 antibody, (CD3 Expanded T cells), and CD8+ T cells expanded with anti-TCRVβ 6-5 antibody (Drug Expanded T cells). The Effector mediators are Granzyme B, IFNɣ, MIP-1α, perforin, and TNFβ. The Chemoattractive mediators are MIP-1b and RANTES.

[0379] FIG.99 is a schematic of the experimental design for the pharmacokinetic (PK) profile and dosing strategy of the multifunctional polypeptide molecule as described herein.

[0380] FIG.100 shows alignment of affinity matured humanized Antibody A-H VL sequences (SEQ ID NOS: 3377-3389, respectively, in order of appearance).

[0381] FIG.101 shows alignment of affinity matured humanized Antibody A-H VH sequences (SEQ ID NOS: 3390-3436, respectively, in order of appearance).

[0382] FIG.102A shows an exemplary embodiment (e.g., BKM0186) of multifunctional molecules comprising a TCRβV-binding moiety and a cytokine polypeptide (e.g., IL2 or IL2-C125A) as described herein. FIG.102B shows an exemplary embodiment of multifunctional molecules comprising a TCRβV- binding moiety and a cytokine polypeptide as described herein. FIGS.102C, 102D, 102E, and 102F show exemplary embodiments of multifunctional molecules comprising a first TCRβV-binding moiety, a second TCRβV-binding moiety, and two cytokine polypeptides as described herein. In some embodiments, the cytokine polypeptide comprises IL-2 or a functional fragment or a functional variant thereof, IL2-C125A or a functional fragment or a functional variant thereof, IL-15 or a functional fragment or a functional variant thereof, IL-7 or a functional fragment or a functional variant thereof, IL- 12 or a functional fragment or a functional variant thereof, or IL-21 or a functional fragment or a functional variant thereof. In embodiments, the cytokine polypeptide further comprises a cytokine receptor. In some embodiments, the cytokine polypeptide comprises IL-15 linked to a IL-15Ra. In some embodiments, the cytokine polypeptide comprises IL-15 linked to a IL-15Ra sushi domain. In some embodiments, the cytokine polypeptide comprises a cytokine dimer. In some embodiments, the cytokine polypeptide comprises an IL-12 beta subunit linked to an IL-12 alpha subunit.

[0383] FIG.103 shows FACS plots showing binding of BKM0186 to different immune cell populations in Human PBMCs.

[0384] FIG.104 shows binding of BKM0186 to pure human T cells expressing either Vβ6 or CD25 (IL- 2Rα) or both.

[0385] FIG.105 shows in vitro concentration-effect relationships for BKM0186-mediated in vitro expansion of Vβ6 T cells and activated (CD25) Vβ6 T cells from human PBMCs at day 5 as a % of total T-Cytotoxic (CD8) and T-helper (CD4) populations. Left graph: T-cytotoxic lymphocytes; right graph: T-helper lymphocytes.WSGR Docket No.53676-769.601

[0386] FIG.106 shows in vitro TCR sequencing. PBMCs were incubated with 100nM of BKM0186 for 5 days and T cells were sequenced for TCR β chain V (TRBV) genes. Compared to unstimulated T cells (grey), BKM0186 selectively expanded T cells bearing TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-5, and TRBV10-3.

[0387] FIG.107A and FIG.107B show a series of graphs (FIG.107A) and a series of FACS plots (FIG.107B) exhibiting activation of CD4+ and CD8+ T cells as assessed by CD25 expression following stimulation with BKM0186, RSV-IL2 and Anti-TCRVβ6 control in solution.

[0388] FIG.108 shows a series of FACS plots demonstrating differentiation of memory T cells mediated by BKM0186 in comparison to unstimulated and the controls RSV-IL2 and anti-TCRVβ6. Upper left quadrant represents Central memory (CM), lower left quadrant represents Effector memory (EM), upper right quadrant represents Naïve (N) and lower right quadrant represents Effector memory RA (TEMRA).

[0389] FIG.109 shows in vitro concentration-effect relationships for BKM0186-induced cytokine release from human PBMCs at day 4 using MSD V-plex human cytokine panel.

[0390] FIG.110 shows BKM0186-mediated killing of human tumor organoids generated from primary, patient-derived tissue from colorectal and NSCLC cancer patients. Vertical bars represent percentage of organoid area reduced relative to isotype control following incubation of organoids with BKM0186 and autologous TILs.

[0391] FIG.111 shows tumor growth curves of mBKM0186-treated EMT6 tumor-bearing mice. Studies were performed in randomized mice with tumor volumes of 80-150 mm3. For all models except MC38, mice were dosed for 3 weeks with a weekly dosing of 0.5-1.5 mg / kg and survival was determined based on 2000 mm3tumor volume end point.

[0392] FIG.112 shows tumor growth curves of mBKM0186-treated mice. Studies were performed in randomized mice with tumor volumes of 80-150 mm3. For all models except MC38, mice were dosed for 4 weeks with a weekly dosing of 1-1.5 mg / kg and survival was determined based on 2000 mm3tumor volume end point. For MC38, mice were given first dose of 3 mg / kg followed by 1 mg / kg for subsequent three weekly (QW) doses.

[0393] FIG.113 shows Kaplan-Meier survival curves of treated mice. Studies were performed in randomized mice with tumor volumes of 80-150 mm3. For all models except MC38, mice were dosed for 4 weeks with a weekly dosing of 1-1.5 mg / kg and survival was determined based on 2000 mm3tumor volume end point. For MC38, mice were given first dose of 3 mg / kg followed by 1 mg / kg for subsequent three weekly (QW) doses.

[0394] FIG.114 shows the experimental design for the tumor rechallenge study. Cured EMT6 tumor bearing mice were rechallenged with EMT6 tumor cells in one flank and CT26 tumor cells in another flank and monitored for tumor growth for 28 days.

[0395] FIG.115 shows the results of the tumor rechallenge study. While the EMT6 tumors were rejected, CT26 tumors grew, suggesting that the memory response against EMT6 tumors likely mediated through mBKM0186 treatment has been established.WSGR Docket No.53676-769.601

[0396] FIG.116 shows immune profiling of T cells in blood and tumor tissue on day 14 post dosing of mBKM0186.

[0397] FIG.117 shows tumor growth curves of EMT6 tumors after weekly (QW) treatment of mice bearing 150 mm3tumors with 1 mg / kg of mBKM0186 with and without depletion of Vβ-specific T cells. Filled Triangles indicate dosing intervals of the depleting antibodies and open Triangles indicate dosing intervals of mBKM0186.

[0398] FIG.118A and FIG.118B show Pharmacokinetic profiles of BKM0186 (FIG.118A) and BKM0281 (FIG.118B) administered single dose IV in cynomolgus monkeys.

[0399] FIG.119A shows T cell expansion following a single IV dose of BKM0186. FIG.119B shows T cell expansion following a single IV dose BKM0281. n=3 Monkeys, n=1 monkey vehicle control.

[0400] FIG.120 shows serum soluble CD25 levels in monkeys administered a single IV dose of BKM0186. Mean values, n=2-3 monkeys per group.

[0401] FIG.121A shows serum IL-6 levels in monkeys administered a single IV dose of BKM0186. FIG.121B shows serum IL-6 levels in monkeys administered a single IV dose of BKM0281. Mean values, n=2-3 monkeys per group.

[0402] FIG.122A shows serum IFN-γ levels in monkeys administered a single IV dose of BKM0186. FIG.122B shows IFN-γ levels in monkeys administered a single IV dose of BKM0281. Mean values, n=2-3 monkeys per group. Mean values, n=2-3 monkeys per group.

[0403] FIG.123 shows in vitro concentration-effect relationships for bispecific-mediated in vitro expansion of Vβ6 T cells.

[0404] FIGs.124A and 124B show exemplary embodiments of multifunctional molecules comprising a TCRβV-binding moiety and a cytokine molecule (e.g., wild-type human IL2 or IL15-IL15R sushi fusion) as described herein. FIGs.124C-124T show exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA) binding moiety, a TCRβV- binding moiety, and at least one cytokine molecule or functional fragment or functional variant thereof as described herein. In some embodiments, a tumor-associated antigen (TAA)-binding moiety and / or a TCRβV-binding moiety are antibody, an antigen binding fragment thereof or an antibody fragment. In some embodiments, an antigen binding fragment or an antibody fragment comprises Fab, Fab′, F(ab′)2, F(ab)2, variable fragment (Fv), domain antibody (dAb), or single chain variable fragment (scFv). In some embodiments, the at least one cytokine molecule or functional fragment or functional variant thereof is selected from the group consisting of interleukin-2 (IL-2) or functional fragment or functional variant thereof, interleukin-7 (IL-7) or functional fragment or functional variant thereof, interleukin-12 (IL-12) or functional fragment or functional variant thereof, interleukin-15 (IL-15) or functional fragment or functional variant thereof, interleukin-18 (IL-18) or functional fragment or functional variant thereof, interleukin-21 (IL-21) or functional fragment or functional variant thereof, or interferon gamma or functional fragment or functional variant thereof, or a combination thereof. In some embodiments, exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA)-binding moiety, a TCRβV-binding moiety, and at least one cytokineWSGR Docket No.53676-769.601 molecule or functional fragment or functional variant thereof as described herein comprises a dimerization module comprising an Fc region comprising N297A mutation. FIGs.124C-124H show exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA)-binding moiety, a TCRβV-binding moiety, and at least one cytokine molecule or functional fragment or functional variant thereof as described herein, wherein the multifunctional molecules comprise an exemplary dimerization module, e.g., an Fc region comprising Knob-in-hole mutations and disulfide bridges. FIGs.124I-124N show exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA)-binding moiety, a TCRβV-binding moiety, and at least one cytokine molecule or functional fragment or functional variant thereof as described herein, wherein the multifunctional molecules comprise an exemplary dimerization module, e.g., an Fc region comprising disulfide bridges, but not comprising Knob-in-hole mutations. FIGs.124O-124T show exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA)-binding moiety, a TCRβV-binding moiety, and at least one cytokine molecule or functional fragment or functional variant thereof as described herein, wherein the multifunctional molecules comprise an exemplary dimerization module, e.g., an Fc region not comprising disulfide bridges nor Knob-in-hole mutations. FIGs.124C- 124E, 124I-124K, and 124O-124Q show exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA)-binding moiety, a TCRβV- binding moiety, and at least one cytokine molecule or functional fragment or functional variant thereof as described herein, wherein the TCRβV-binding moiety comprises an scFv. FIGs.124F-124H, 124L- 124N, and 124R-124T show exemplary embodiments of multifunctional molecules, e.g., multifunctional molecules comprising a tumor-associated antigen (TAA)-binding moiety, a TCRβV-binding moiety, and at least one cytokine molecule or functional fragment or functional variant thereof as described herein, wherein the TCRβV-binding moiety comprises an Fab. In some embodiments, exemplary embodiments of multifunctional molecules comprises an antibody molecule, an antigen binding domain thereof, or a functional fragment or variant that binds to a TCRαV region, in lieu of an antibody molecule that binds to a T cell receptor beta variable region (TCRβV). In some embodiments, exemplary embodiments of multifunctional molecules comprises an antibody molecule, an antigen binding domain thereof, or a functional fragment or variant that binds to DLL-3 as an a-TAA.

[0405] FIG.125A-125B show pharmacokinetics of CD3 x gp75 bispecific molecule (BNM0092), which is an exemplary embodiment of the CD3 binding agent as provided herein. gp75 (TA99 / TRP1)-bound FLAG-tagged CD3 x gp75 bispecific molecule (BNM0092) was detected in plasma samples between 0 and 120 hours. Half-time, AUC, and Cmax of CD3 x gp75 bispecific molecule (BNM0092) was determined (FIG.125A). Biotinylated Mouse monoclonal anti-FLAG antibody was used as primary detection for gp75 (TA99 / TRP1)-bound FLAG-tagged CD3 x gp75 bispecific molecule (BNM0092), SULFO-Streptavidin was used as secondary detection to amplify the primary signal (FIG.125B)

[0406] FIGs.126A-126B show tumor volume (FIG.126A) and probability of survival (FIG.126B) in tumor-bearing mice dosed I.P. with vehicle, CD3 x gp75 bispecific molecule (BNM0092) 2mg / kgWSGR Docket No.53676-769.601 biweekly for 4 weeks, anti-TCRVβ 13-3 (BKM0307) at 1mg / kg once, or anti-TCRVβ 13-3 (BKM0307) at 1mg / kg once and CD3 x gp75 bispecific molecule (BNM0092) at 2mg / kg biweekly for 4 weeks.

[0407] FIGs.127A-127B show tumor volume (FIG.127A) and probability of survival (FIG.127B) in tumor-bearing mice dosed I.P. with vehicle, CD3 x gp75 bispecific molecule (BNM0092) 2mg / kg biweekly for 3 weeks, anti-TCRVβ 13-3 (BKM0307) at 1mg / kg three times on day 0, 10, 20, or anti- TCRVβ 13-3 (BKM0307) at 1mg / kg three times on day 0,10, 20 and CD3 x gp75 bispecific molecule (BNM0092) at 2mg / kg biweekly for 3 weeks.

[0408] FIGs.128A-128B show tumor volume (FIG.128A) and probability of survival (FIG.128B) in tumor-bearing mice dosed I.P. with vehicle; anti-TCRVβ 13-3 (BKM0307) at 1mg / kg biweekly for 4 weeks; CD3 x gp75 bispecific molecule (BNM0092) at 2mg / kg biweekly for 4 weeks; CD3 x gp75 bispecific molecule (BNM0092) at 0.5mg / kg biweekly for 4 weeks; anti-TCRVβ 13-3 (BKM0307) at 1mg / kg and CD3 x gp75 bispecific molecule (BNM0092) at 2mg / kg biweekly for 4 weeks; anti-TCRVβ 13-3 (BKM0307) at 1mg / kg and CD3 x gp75 bispecific molecule (BNM0092) at 0.5mg / kg biweekly for 4 weeks. DETAILED DESCRIPTION DEFINITION

[0409] Certain specific details of this description are set forth in order to provide a thorough understanding of various embodiments. However, one skilled in the art will understand that the present disclosure may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring descriptions of the embodiments.

[0410] Unless the context requires otherwise, throughout the specification and claims which follow, the word “comprise” and variations thereof, such as, “comprises” and “comprising” are to be construed in an open, inclusive sense, that is, as “including, but not limited to.” Further, headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed disclosure.

[0411] As used in this specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the content clearly dictates otherwise. 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.”

[0412] It should also be noted that the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise.

[0413] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below.

[0414] The term “about” when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or in some instances ±10%, or in some instancesWSGR Docket No.53676-769.601 ±5%, or in some instances ±1%, or in some instances ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. 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.

[0415] The term “acquire” or “acquiring” as the terms are used herein, refer to obtaining possession of a physical entity (e.g., a sample, a polypeptide, a nucleic acid, or a sequence), or a value, e.g., a numerical value, by “directly acquiring” or “indirectly acquiring” the physical entity or value. “Directly acquiring” means performing a process (e.g., performing a synthetic or analytical method) to obtain the physical entity or value. “Indirectly acquiring” refers to receiving the physical entity or value from another party or source (e.g., a third party laboratory that directly acquired the physical entity or value). Directly acquiring a physical entity includes performing a process that includes a physical change in a physical substance, e.g., a starting material. Directly acquiring a value includes performing a process that includes a physical change in a sample or another substance, e.g., performing an analytical process which includes a physical change in a substance, e.g., a sample.

[0416] “Antibody molecule” as used herein refers to a protein, e.g., an immunoglobulin chain or fragment thereof, comprising at least one immunoglobulin variable domain structure and / or sequence. An antibody molecule encompasses antibodies (e.g., full-length antibodies) and antibody fragments. In some embodiments, 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), single domain antibody, single chain variable fragment (scFv), a nanobody (VHH), or a camelid single domain antibody. 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., domain antibody, single chain, Fab, Fab’, and F(ab’)2 fragments, single chain variable fragments (scFvs), a single domain antibody, a nanobody (VHH), a camelid single domain antibody. In some embodiments, the antibody molecule is an antibody mimetic. In some embodiments, the antibody molecule is, or comprises, an antibody-like framework or scaffold, such as, fibronectins, ankyrin repeats (e.g., designed ankyrin repeat proteins (DARPins)), avimers, affibody affinity ligands, anticalins, or affilin molecules.WSGR Docket No.53676-769.601

[0417] The term “human-like antibody molecule” as used herein refers to a humanized antibody molecule, human antibody molecule or an antibody molecule having at least 95% sequence identity with a non-murine germline framework region, e.g., FR1, FR2, FR3 and / or FR4. In some embodiments, the human-like antibody molecule comprises a framework region having at least 95% sequence identity to a human germline framework region, e.g., a FR1, FR2, FR3 and / or FR4 of a human germline framework region. In some embodiments, the human-like antibody molecule is a recombinant antibody. In some embodiments, the human-like antibody molecule is a humanized antibody molecule. In some embodiments, the human-like antibody molecule is human antibody molecule. In some embodiments, the human-like antibody molecule is a phage display or a yeast display antibody molecule. In some embodiments, the human-like antibody molecule is a chimeric antibody molecule. In some embodiments, the human-like antibody molecule is a CDR grafted antibody molecule.

[0418] 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.

[0419] 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 sequence 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.

[0420] “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.

[0421] 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. InWSGR Docket No.53676-769.601 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, 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.

[0422] 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.

[0423] “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.

[0424] 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.

[0425] 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 labelingWSGR Docket No.53676-769.601 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.

[0426] 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 of genomic, cDNA, semisynthetic, or synthetic origin which either does not occur in nature or is linked to another polynucleotide in a non-natural arrangement.

[0427] 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. An isolated polynucleotide (ribonucleic acid (RNA), deoxyribonucleic acid (DNA)), or polypeptide is free of the genes / nucleic acids or sequences / amino acids that flank it in its naturally-occurring state.

[0428] The compositions and methods of the present disclosure 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% 99%, 99.5%, 99.9%, or 100% sequence identity to a reference sequence, e.g., a sequence provided herein. 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% 99%, 99.5%, 99.9%, or 100% sequence identity to a reference sequence, e.g., a sequence provided herein.WSGR Docket No.53676-769.601

[0429] 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. In some embodiments, a TCRβV variant can bind to TCRα and form a TCR α:β complex.

[0430] 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.

[0431] The term “functional fragment” refers to a polypeptide that has a partial amino acid sequence of a reference amino acid sequence, and is capable of having one or more activities of the reference amino acid sequence. In some embodiments, the functional fragment comprises at least about 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% 99%, 99.5%, or 99.9% amino acid sequence of a reference amino acid sequence. In some embodiments, the functional fragment comprises an amino acid sequence that has at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 amino acid deletion from the reference amino acid sequence. In some embodiments, the functional fragment comprises an amino acid sequence that has at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 350, 400, 450, or 500 amino acids of the reference amino acid sequence.

[0432] Calculations of homology or sequence identity between sequences (the terms are used interchangeably herein) are performed as follows. 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 of 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”).

[0433] 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. 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 hasWSGR Docket No.53676-769.601 been incorporated into the GAP program in the GCG software package (available at http: / / www.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 http: / / www.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.

[0434] 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. 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 present disclosure. 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 present disclosure. 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.

[0435] It is understood that the molecules of the present disclosure may have additional conservative or non-essential amino acid substitutions, which do not have a substantial effect on their functions.

[0436] 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 polymer 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.

[0437] 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).WSGR Docket No.53676-769.601

[0438] 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.

[0439] As used herein, the term “mutation” refers to an alteration in the nucleotide sequence of the genome of an organism, virus, or extrachromosomal DNA. In some embodiments, the mutation may be a large-scale mutation, such as amplifications (or gene duplications) or repetitions of a chromosomal segment, deletions of large chromosomal regions, chromosomal rearrangements (e.g., chromosomal translocations, chromosomal inversions, non-homologous chromosomal crossover, and interstitial deletions), and loss of heterozygosity. In some embodiments, the mutation may be a small-scale mutation, such as insertions, deletions, and substitution mutations. As used herein, the term “substitution mutation” refers to the transition that exchange a single nucleotide for another.

[0440] “Interleukin-2” also known as IL2, IL-2, IL 2, TCGF, lymphokine, and interleukin 2, as referred to herein, includes any of the recombinant or naturally-occurring forms of IL-2 or variants or homologs thereof that have or maintain IL-2 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 IL-2. In some embodiments, IL-2 is substantially identical to the protein identified by the UniProt reference number P60568 or a variant or homolog having substantial identity thereto.

[0441] “DLL3” also known as DLL3, SCDO1, pu, pudgy, delta like canonical Notch ligand 3, and Delta- like 3 (Drosophila), as referred to herein, refers to an inhibitory protein of Notch signaling that is involved in embryonic patterning and has been implicated in neuroendocrine cell development. DLL3, as referred to herein, includes any of the recombinant or naturally-occurring forms of DLL3 or variants or homologs thereof. 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 DLL3. In some embodiments, DLL3 is substantially identical to the protein identified by the UniProt reference number Q9NYJ7 or a variant or homolog having substantial identity thereto.

[0442] “CLDN18”, also known as Claudin-18, Surfactant Associated Protein J, SFTPJ, Surfactant Associated 5, SFTA5, refers to a member of the claudin family. Claudins are integral membrane proteins and components of tight junction strands. CLDN18 localizes to cellular tight junctions in both mouse and human. CLDN18 has two isoforms, CLDN18.1, expressed specifically in the lung, and CLDN18.2, as referred to herein, expressed in differentiated cells of the gastric mucosa. CLDN18.2, as referred to herein, includes any of the recombinant or naturally-occurring forms of CLDN18.2 or variants orWSGR Docket No.53676-769.601 homologs thereof. 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 CLDN18.2. In some embodiments, CLDN18.2 is substantially identical to the protein identified by the UniProt reference number P56856-2 or a variant or homolog having substantial identity thereto. Anti-TCRβV antibodies Human T cell receptor (TCR) complex

[0443] TCR is a disulfide-linked membrane-anchored heterodimeric protein normally consisting of the highly variable alpha (α) and beta (β) chains expressed as part of a complex with the invariant CD3 chain molecules. TCR on αβ T cells is formed by a heterodimer of one alpha chain and one beta chain. Each alpha or beta chain consists of a constant domain and a highly variable domain classified as the Immunoglobulin superfamily (IgSF) fold. The TCRβV chains can be further classified into subfamilies. Despite their high structural and functional homology, the amino acid sequence homology in the TRBV genes is very low. Only 4 amino acids out of approximately 95 are identical while 10 additional amino acids are conserved among all subfamilies. Nevertheless, TCRs formed between alpha and beta chains of highly diverse sequences show a remarkable structural homology and elicit a similar function, e.g., activation of T cells.

[0444] T cell receptors (TCR) can be found on the surface of T cells. TCRs recognize antigens, e.g., peptides, presented on, e.g., bound to, major histocompatibility complex (MHC) molecules on the surface of cells, e.g., antigen-presenting cells. TCRs are heterodimeric molecules and can comprise an alpha chain, a beta chain, a gamma chain or a delta chain. TCRs comprising an alpha chain and a beta chain are also referred to as TCRαβ. The TCR beta chain consists of the following regions (also known as segments): variable (V), diversity (D), joining (J) and constant (C) (see Mayer G. and Nyland J. (2010) Chapter 10: Major Histocompatibility Complex and T-cell Receptors-Role in Immune Responses. In: Microbiology and Immunology on-line, University of South Carolina School of Medicine). The TCR alpha chain consists of V, J and C regions. The rearrangement of the T-cell receptor (TCR) through somatic recombination of V (variable), D (diversity), J (joining), and C (constant) regions is a defining event in the development and maturation of a T cell. TCR gene rearrangement takes place in the thymus.

[0445] TCRs can comprise a receptor complex, known as the TCR complex, which comprises a TCR heterodimer comprising of an alpha chain and a beta chain, and dimeric signaling molecules, e.g., CD3 co-receptors, e.g., CD3δ / ε, and / or CD3γ / ε.

[0446] As used herein, the term “T cell receptor beta variable chain” or “TCRβV,” refers 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 TCRβWSGR Docket No.53676-769.601 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, a TCRβ V29 subfamily, a TCRβ V1 subfamily, a TCRβ V17 subfamily, a TCRβ V21 subfamily, a TCRβ V23 subfamily, or a TCRβ V26 subfamily, as well as family members of said subfamilies, and variants thereof (e.g., a structural or functional variant thereof). 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, or a variant thereof, e.g., a variant having 85%, 90%, 95%, 99% or more identity the naturally-occurring sequence. TCRβ V6-5*01 is also known as TRBV65; TCRBV6S5; TCRBV13S1, or TCRβ V13.1. 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. In some embodiments, TCRβ V6-5*01 is encoded by the nucleic acid sequence of SEQ ID NO: 43, or a sequence having 85%, 90%, 95%, 99% or more identity thereof. In some embodiments, TCRβ V6-5*01 comprises the amino acid sequence of SEQ ID NO: 44, or a sequence having 85%, 90%, 95%, 99% or more identity thereof.

[0447] SEQ ID NO: 43 ATGAGCATCGGCCTCCTGTGCTGTGCAGCCTTGTCTCTCCTGTGGGCAGGTCCAGTGAATGC TGGTGTCACTCAGACCCCAAAATTCCAGGTCCTGAAGACAGGACAGAGCATGACACTGCAG TGTGCCCAGGATATGAACCATGAATACATGTCCTGGTATCGACAAGACCCAGGCATGGGGC TGAGGCTGATTCATTACTCAGTTGGTGCTGGTATCACTGACCAAGGAGAAGTCCCCAATGGC TACAATGTCTCCAGATCAACCACAGAGGATTTCCCGCTCAGGCTGCTGTCGGCTGCTCCCTC CCAGACATCTGTGTACTTCTGTGCCAGCAGTTACTC

[0448] SEQ ID NO: 44 MSIGLLCCAALSLLWAGPVNAGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMG LRLIHY-SVGAGITDQGEVPNGYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSY TCR beta V (TCRβV)

[0449] Diversity in the immune system enables protection against a huge array of pathogens. Since the germline genome is limited in size, diversity is achieved not only by the process of V(D)J recombination but also by junctional (junctions between V-D and D-J segments) deletion of nucleotides and addition of pseudo-random, non-templated nucleotides. The TCR beta gene undergoes gene arrangement to generate diversity.

[0450] The TCR V beta repertoire varies between individuals and populations because of, e.g., 7 frequently occurring inactivating polymorphisms in functional gene segments and a large insertion / deletion-related polymorphism encompassing 2 V beta gene segments.

[0451] Provided herein are, inter alia, antibody molecules and fragments thereof, that bind, e.g., specifically bind, to a human TCR beta V chain (TCRβV), e.g., a TCRβV gene family (also referred to asWSGR Docket No.53676-769.601 a group), e.g., a TCRβV subfamily (also referred to as a subgroup), e.g., as described herein. TCR beta V families and subfamilies are known in the art, e.g., as described in Yassai et al., (2009) Immunogenetics 61(7)pp:493-502; Wei S. and Concannon P. (1994) Human Immunology 41(3) pp: 201-206. The antibodies described herein can be recombinant antibodies, e.g., recombinant non-murine antibodies, e.g., recombinant human or humanized antibodies.

[0452] The terms TCRBV, TCRVB, TRBV, TCRβV, TCRVβ or TRβV are used interchangeably herein and refer to a TCR beta V chain, e.g., as described herein.

[0453] In some embodiments, provided herein is an anti-TCRβV antibody molecule that binds to human TCRβV, e.g., a TCRβV family, e.g., gene family or a variant thereof. In some embodiments a TCRBV gene family comprises one or more subfamilies, e.g., as described herein, e.g., Table 8A or Table 8B. In some embodiments, the TCRβV gene family comprises: 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, a TCRβ V29 subfamily, a TCRβ V1 subfamily, a TCRβ V17 subfamily, a TCRβ V21 subfamily, a TCRβ V23 subfamily, or a TCRβ V26 subfamily.

[0454] In some embodiments, TCRβ V6 subfamily is also known as TCRβ V13.1. 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, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-9*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-8*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-5*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-2*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-3*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-1*01, or a variant thereof.

[0455] In some embodiments, TCRβ V6 comprises TCRβ V6-5*01, or a variant thereof. In some embodiments, TCRβ V6, e.g., TCRβ V6-5*01, is recognized, e.g., bound, by SEQ ID NO: 1 and / or SEQ ID NO: 2. In some embodiments, TCRβ V6, e.g., TCRβ V6-5*01, is recognized, e.g., bound, by SEQ ID NO: 9 and / or SEQ ID NO: 10. In some embodiments, TCRβ V6 is recognized, e.g., bound, by SEQ ID NO: 9 and / or SEQ ID NO: 11.

[0456] In some embodiments, TCRβ V10 subfamily is also known as TCRβ V12. In some embodiments, the TCRβ V10 subfamily comprises: TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-3*01 or TCRβ V10-2*01, or a variant thereof.WSGR Docket No.53676-769.601

[0457] In some embodiments, TCRβ V12 subfamily is also known as TCRβ V8.1. In some embodiments, the TCRβ V12 subfamily comprises: TCRβ V12-4*01, TCRβ V12-3*01, or TCRβ V12-5*01, or a variant thereof. In some embodiments, TCRβ V12 is recognized, e.g., bound, by SEQ ID NO: 15 and / or SEQ ID NO: 16. In some embodiments, TCRβ V12 is recognized, e.g., bound, by any one of SEQ ID NOs 23-25, and / or any one of SEQ ID NO: 26-30:

[0458] In some embodiments, the TCRβ V5 subfamily is chosen from: TCRβ V5-5*01, TCRβ V5-6*01, TCRβ V5-4*01, TCRβ V5-8*01, TCRβ V5-1*01, or a variant thereof.

[0459] In some embodiments, the TCRβ V7 subfamily comprises TCRβ V7-7*01, TCRβ V7-6*01, TCRβ V7 -8*02, TCRβ V7 -4*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*01, TCRβ V7-3*01, TCRβ V7-9*03, or TCRβ V7-9*01, or a variant thereof.

[0460] In some embodiments, the TCRβ V11 subfamily comprises: TCRβ V11-1*01, TCRβ V11-2*01 or TCRβ V11-3*01, or a variant thereof. In some embodiments, the TCRβ V14 subfamily comprises TCRβ V14*01, or a variant thereof. In some embodiments, the TCRβ V16 subfamily comprises TCRβ V16*01, or a variant thereof. In some embodiments, the TCRβ V18 subfamily comprises TCRβ V18*01, or a variant thereof. In some embodiments, the TCRβ V9 subfamily comprises TCRβ V9*01 or TCRβ V9*02, or a variant thereof. In some embodiments, the TCRβ V13 subfamily comprises TCRβ V13*01, or a variant thereof. In some embodiments, the TCRβ V4 subfamily comprises TCRβ V4-2*01, TCRβ V4-3*01, or TCRβ V4-1*01, or a variant thereof. In some embodiments, the TCRβ V3 subfamily comprises TCRβ V3-1*01, or a variant thereof. In some embodiments, the TCRβ V2 subfamily comprises TCRβ V2*01, or a variant thereof. In some embodiments, the TCRβ V15 subfamily comprises TCRβ V15*01, or a variant thereof. In some embodiments, the TCRβ V30 subfamily comprises TCRβ V30*01, or TCRβ V30*02, or a variant thereof. In some embodiments, the TCRβ V19 subfamily comprises TCRβ V19*01, or TCRβ V19*02, or a variant thereof. In some embodiments, the TCRβ V27 subfamily comprises TCRβ V27*01, or a variant thereof. In some embodiments, the TCRβ V28 subfamily comprises TCRβ V28*01, or a variant thereof. In some embodiments, the TCRβ V24 subfamily comprises TCRβ V24-1*01, or a variant thereof. In some embodiments, the TCRβ V20 subfamily comprises TCRβ V20-1*01, or TCRβ V20-1*02, or a variant thereof. In some embodiments, the TCRβ V25 subfamily comprises TCRβ V25-1*01, or a variant thereof. In some embodiments, the TCRβ V29 subfamily comprises TCRβ V29-1*01, or a variant thereof.

[0461] Exemplary amino acid sequences for TCRβV subfamily members can be found on the ImMunoGeneTics Information System website: http: / / www.imgt.org / , or in a similar resource.

[0462] Current bispecific constructs designed to redirect T cells to promote tumor cell lysis for cancer immunotherapy typically utilize antibody fragments (Fab, scFv, VH, single domain antibody, etc.) that are derived from monoclonal antibodies (mAb) directed against the CD3e subunit of the T cell receptor (TCR). However, there are limitations to this approach which may prevent the full realization of the therapeutic potential for such bispecific constructs. Previous studies have shown that even low “activating” doses of anti-CD3e mAb can cause long-term T cell dysfunction and exert immunosuppressive effects. In addition, anti-CD3e mAbs have been associated with side effects thatWSGR Docket No.53676-769.601 result from massive T cell activation. The large number of activated T cells secrete substantial amounts of cytokines, the most important of which is Interferon gamma (IFNγ). This excess amount of IFNγ in turn activates macrophages which then overproduce proinflammatory cytokines such as IL-1beta, IL-6, IL-10 and TNF-alpha, causing a “cytokine storm” known as the cytokine release syndrome (CRS) (Shimabukuro-Vornhagen et al., J Immunother Cancer.2018 Jun 15;6(1):56, herein incorporated by reference in its entirety). Thus, the need exists for developing antibodies that are capable of binding and activating only a subset of effector T cells, e.g., to re-duce the CRS and / or neurotoxicity (NT).

[0463] Described herein are molecules targeting the TCRβV chain of TCR and methods thereof. Without wishing to be bound by theory, such molecules are capable of binding, activating, and / or expanding only a subset of T cells, avoiding or reducing CRS and / or NT and minimizing potential immunosuppressive effects of anti-CD3 mAbs.

[0464] Described herein is a class of antibodies, i.e., anti-TCRβV antibody molecules as described herein, which despite having low sequence similarity (e.g., low sequence identity among the different antibody molecules that recognize different TCRβV subfamilies), recognize a structurally conserved, yet sequence-wise variable, region, e.g., domain, on the TCRβV protein and have a similar function (e.g., activation of T cells and a similar cytokine profile as described herein). Thus, the anti-TCRβV antibody molecules as described herein share a structure-function relationship.

[0465] Without wishing to be bound by theory, in some embodiments, the anti-TCRβV antibody molecules as described herein bind to an outward facing epitope of a TCRβV protein when it is in a complex with a TCRalpha protein. In some embodiments, the anti-TCRβV antibody molecules as described herein recognize (e.g., bind to), a domain (e.g., an epitope) on the TCRβV protein that is: (1) structurally conserved among different TCRβV subfamilies; and (2) has minimal sequence identity among the different TCRβV subfamilies. TCRβV proteins from the different TCRBV subfamilies share minimal sequence similarity. However, TCRβV proteins which have minimal sequence similarity, share a similar 3D conformation and structure.

[0466] The alignment of TCRBV amino acid sequences in FIG.3 underscores the diversity of TCR sequences. In particular, the TRBV sequences from different subfamilies are considerably different from each other.

[0467] In some embodiments, the anti-TCRβV antibody molecules as described herein do not recognize, e.g., bind to, an interface of a TCRβV:TCRalpha complex. In some embodiments, the anti-TCRβV antibody molecules as described herein do not recognize, e.g., bind to, a constant region of a TCRβV protein. An exemplary antibody that binds to a constant region of a TCRBV region is JOVI.1 as de- scribed in Viney et al., (Hybridoma.1992 Dec;11(6):701-13). In some embodiments, the anti-TCRβV antibody molecules as described herein do not recognize, e.g., bind to, one or more (e.g., all) of a complementarity determining region (e.g., CDR1, CDR2 and / or CDR3) of a TCRβV protein.

[0468] Provided herein are, inter alia, antibody molecules directed to the variable chain of the beta subunit of TCR (TCRβV) which bind and, e.g., activate a subset of T cells. The anti-TCRβV antibody molecules as described herein result in lesser or no production of cytokines associated with CRS, e.g., IL-WSGR Docket No.53676-769.601 6, IL-1beta, IL-10 and TNF alpha; and enhanced and / or delayed production of IL-2 and IFNγ. In some embodiments, the anti-TCRβV antibodies as described herein have a cytokine profile, e.g., as described herein, which differs from a cytokine profile of a T cell engager that binds to a receptor or molecule other than a TCRβV region (“a non-TCRβV-binding T cell engager”). In some embodiments, the non-TCRβV- binding T cell engager comprises an antibody that binds to a CD3 molecule (e.g., CD3 epsilon (CD3e) molecule); or a TCR alpha (TCRα) molecule. In some embodiments, the non-TCRβV-binding T cell engager is an OKT3 antibody or an SP34-2 antibody.

[0469] In some embodiments, the anti-TCRβV antibodies as described herein result in expansion of TCRβV+ T cells, e.g., a subset of memory effector T cells known as TEMRA. Without wishing to be bound by theory, it is believed that in some embodiments, TEMRA cells can promote tumor cell lysis but not CRS. Accordingly, provided herein are methods of making said anti-TCRβV antibody molecules and uses thereof. Also described herein are multispecific molecules, e.g., bispecific molecules comprising said anti-TCRβV antibody molecules. In some embodiments, compositions comprising anti-TCRβV antibody molecules of the present disclosure, can be used, e.g., to: (1) activate and redirect T cells to promote tumor cell lysis for cancer immuno-therapy; and / or (2) expand TCRβV+ T cells. In some embodiments, compositions comprising anti-TCRβV antibody molecules as described herein limit the harmful side-effects of CRS and / or NT, e.g., CRS and / or NT associated with anti-CD3e targeting.

[0470] In some embodiments, the anti-TCRβV antibody molecule binds to one or more of TRBV2, TRBV3-1, TRBV4-1, TRBV4-2, TRBV4-3, TRBV5-1, TRBV5-4, TRBV5-5, TRBV5-6, TRBV5-8, TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-4, TRBV6-5, TRBV6-6, TRBV6-8, TRBV6-9, TRBV7-2, TRBV7-3, TRBV7-4, TRBV7-6, TRBV7-7, TRBV7-8, TRBV7-9, TRBV9, TRBV10-1, TRBV10-2, TRBV10-3, TRBV11-1, TRBV11-2, TRBV11-3, TRBV12-3, TRBV12-4, TRBV12-5, TRBV13, TRBV14, TRBV15, TRBV16, TRBV18, TRBV19, TRBV20-1, TRBV24-1, TRBV25-1, TRBV27, TRBV28, TRBV29-1 and TRBV30. In some embodiments, the anti-TCRβV antibody molecule binds to one or more of TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-4, TRBV6-5, TRBV6-6, TRBV6-8 and TRBV6-9. In some embodiments, the anti-TCRβV antibody molecule is an anti-TRBV2, anti-TRBV3-1, anti-TRBV4-1, anti-TRBV4-2, anti-TRBV4-3, anti-TRBV5-1, anti-TRBV5-4, anti-TRBV5-5, anti- TRBV5-6, anti-TRBV5-8, anti-TRBV6-1, anti-TRBV6-2, anti-TRBV6-3, anti-TRBV6-4, anti-TRBV6-5, anti-TRBV6-6, anti-TRBV6-8, anti-TRBV6-9, anti-TRBV7-2, anti-TRBV7-3, anti-TRBV7-4, anti- TRBV7-6, anti-TRBV7-7, anti-TRBV7-8, anti-TRBV7-9, anti-TRBV9, anti-TRBV10-1, anti-TRBV10- 2, anti-TRBV10-3, anti-TRBV11-1, anti-TRBV11-2, anti-TRBV11-3, anti-TRBV12-3, anti-TRBV12-4, anti-TRBV12-5, anti-TRBV13, anti-TRBV14, anti-TRBV15, anti-TRBV16, anti-TRBV18, anti- TRBV19, anti-TRBV20-1, anti-TRBV24-1, anti-TRBV25-1, anti-TRBV27, anti-TRBV28, anti- TRBV29-1, or anti-TRBV30. Exemplary anti-TCRβV antibody molecules and the corresponding TCRβV subfamilies recognized by said anti-TCRβV antibody molecules are disclosed in Table 10A.

[0471] In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV2, TRBV3- 1, TRBV4-1, TRBV4-2, TRBV4-3, TRBV5-1, TRBV5-4, TRBV5-5, TRBV5-6, TRBV5-8, TRBV6-1, TRBV6-2, TRBV6-3, TRBV6-4, TRBV6-5, TRBV6-6, TRBV6-8, TRBV6-9, TRBV7-2, TRBV7-3,WSGR Docket No.53676-769.601 TRBV7-4, TRBV7-6, TRBV7-7, TRBV7-8, TRBV7-9, TRBV9, TRBV10-1, TRBV10-2, TRBV10-3, TRBV11-1, TRBV11-2, TRBV11-3, TRBV12-3, TRBV12-4, TRBV12-5, TRBV13, TRBV14, TRBV15, TRBV16, TRBV18, TRBV19, TRBV20-1, TRBV24-1, TRBV25-1, TRBV27, TRBV28, TRBV29-1 or TRBV30. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-1. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-2. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-3. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-4. In some embodiments, the anti- TCRβV antibody molecule binds specifically to TRBV6-5. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-6. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-8. In some embodiments, the anti-TCRβV antibody molecule binds specifically to TRBV6-9.

[0472] In some embodiments, the anti-TCRβV antibody molecule does not bind to TCRβ V12, or binds to TCRβ V12 with an affinity and / or binding specificity that is less than (e.g., less than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the 16G8 murine antibody or a humanized version thereof as de-scribed in US Patent 5,861,155.

[0473] In some embodiments, the anti-TCRβV antibody molecule binds to TCRβ V12 with an affinity and / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the 16G8 murine antibody or a humanized version thereof as described in US Patent 5,861,155.

[0474] In some embodiments, the anti-TCRβV antibody molecule binds to a TCRβV region other than TCRβ V12 (e.g., TCRβV region as described herein, e.g., TCRβ V6 subfamily (e.g., TCRβ V6-5*01) with an affinity and / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the 16G8 murine antibody or a humanized version thereof as de-scribed in US Patent 5,861,155.

[0475] In some embodiments, the anti-TCRβV antibody molecule does not comprise the CDRs of the Antibody B murine antibody.

[0476] In some embodiments, the anti-TCRβV antibody molecule does not bind to TCRβ V5-5*01 or TCRβ V5-1*01, or binds to TCRβ V5-5*01 or TCRβ V5-1*01 with an affinity and / or binding specificity that is less than (e.g., less than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the TM23 murine antibody or a humanized version thereof as described in US Patent 5,861,155.

[0477] In some embodiments, the anti-TCRβV antibody molecule binds to TCRβ V5-5*01 or TCRβ V5- 1*01with an affinity and / or binding specificity that is greater than (e.g., greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the TM23 murine antibody or a humanized version thereof as de-scribed in US Patent 5,861,155.

[0478] In some embodiments, the anti-TCRβV antibody molecule binds to a TCRβV region other than TCRβ V5-5*01 or TCRβ V5-1*01 (e.g., TCRβV region as described herein, e.g., TCRβ V6 subfamily (e.g., TCRβ V6-5*01) with an affinity and / or binding specificity that is greater than (e.g., greater thanWSGR Docket No.53676-769.601 about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or about 2-, 5-, or 10- fold) the affinity and / or binding specificity of the TM23 murine antibody or a humanized version thereof as described in US Patent 5,861,155.

[0479] In some embodiments, the anti-TCRβV antibody molecule does not comprise the CDRs of the TM23 murine antibody.

[0480] In some embodiments, the light or the heavy chain variable framework (e.g., the region encompassing at least FR1, FR2, FR3, and optionally FR4) of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule can be chosen from: (a) a light or heavy chain variable framework including at least 80%, 85%, 87% 90%, 92%, 93%, 95%, 97%, 98%, or 100% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (b) a light or heavy chain variable framework including from 20% to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (c) a non-human framework (e.g., a rodent framework); or (d) a non-human framework that has been modified, e.g., to remove antigenic or cytotoxic determinants, e.g., deimmunized, or partially humanized. In some embodiments, the light or heavy chain variable framework region (particularly FR1, FR2 and / or FR3) includes a light or heavy chain variable framework sequence at least 70, 75, 80, 85, 87, 88, 90, 92, 94, 95, 96, 97, 98, 99% identical or identical to the frameworks of a VL or VH segment of a human germline gene.

[0481] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a heavy chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more changes, e.g., amino acid substitutions or deletions, from an amino acid sequence of any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, e.g., the amino acid sequence of the FR region in the entire variable region, e.g., SEQ ID NO: 9.

[0482] Alternatively, or in combination with the heavy chain substitutions described herein, the anti- TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more amino acid changes, e.g., amino acid substitutions or deletions, from an amino acid sequence of any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, e.g., the amino acid sequence of the FR region in the entire variable region, e.g., SEQ ID NO: 10 or SEQ ID NO: 11.

[0483] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes one, two, three, or four heavy chain framework regions, or a sequence substantially identical thereto.

[0484] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes one, two, three, or four light chain framework regions, or a sequence substantially identical thereto.WSGR Docket No.53676-769.601

[0485] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework region 1 of A-H.1 or A-H.2.

[0486] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework region 2 of A-H.1 or A-H.2.

[0487] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework region 3 of A-H.1 or A-H.2.

[0488] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework region 4 of A-H.1 or A-H.2.

[0489] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain comprising a framework region, e.g., framework region 1 (FR1), comprising a change, e.g., a substitution (e.g., a conservative substitution) at position 10 according to Kabat numbering. In some embodiments, the FR1 comprises a Phenylalanine at position 10, e.g., a Serine to Phenyalanine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0490] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain comprising a framework region, e.g., framework region 2 (FR2), comprising a change, e.g., a substitution (e.g., a conservative substitution) at a position as described herein according to Kabat numbering. In some embodiments, FR2 comprises a Histidine at position 36, e.g., a substitution at position 36 according to Kabat numbering, e.g., a Tyrosine to Histidine substitution. In some embodiments, FR2 comprises an Alanine at position 46, e.g., a substitution at position 46 according to Kabat numbering, e.g., an Arginine to Alanine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0491] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain comprising a framework region, e.g., framework region 3 (FR3), comprising a change, e.g., a substitution (e.g., a conservative substitution) at a position as described herein according to Kabat numbering. In some embodiments, FR3 comprises a Phenyalanine at position 87, e.g., a substitution at position 87 according to Kabat numbering, e.g., a Tyrosine to Phenyalanine substitution. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0492] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain comprising: (a) a framework region 1 (FR1) comprising a Phenylalanine at position 10, e.g., a substitution at position 10 according to Kabat numbering, e.g., a Serine to Phenyalanine substitution; (b) a framework region 2 (FR2) comprising a Histidine at position 36, e.g., a substitution at position 36 according to Kabat numbering, e.g., a Tyrosine to Histidine substitution, and a Alanine at position 46, e.g., a substitution at position 46 according to Kabat numbering, e.g., a Arginine to Alanine substitution; and (c) a framework region 3 (FR3) comprising a Phenylalanine at position 87, e.g., a substitution at position 87 according to KabatWSGR Docket No.53676-769.601 numbering, e.g., a Tyrosine to Phenyalanine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 10. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0493] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain comprising: (a) a framework region 2 (FR2) comprising a Histidine at position 36, e.g., a substitution at position 36 according to Kabat numbering, e.g., a Tyrosine to Histidine substitution, and a Alanine at position 46, e.g., a substitution at position 46 according to Kabat numbering, e.g., a Arginine to Alanine substitution; and (b) a framework region 3 (FR3) comprising a Phenylalanine at position 87, e.g., a substitution at position 87 according to Kabat numbering, e.g., a Tyrosine to Phenyalanine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 11. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0494] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a light chain variable domain comprising: (a) a framework region 1 (FR1) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) positions as described herein according to Kabat numbering, ; (b) a framework region 2 (FR2) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) position as described herein according to Kabat numbering and (c) a framework region 3 (FR3) comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more (e.g., all) position as described herein according to Kabat numbering. In some embodiments, the substitution is relative to a human germline light chain framework region sequence.

[0495] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 1 of A-H.1 or A-H.2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 2 of A-H.1 or A-H.2, e.g. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 3 of A-H.1 or A-H.2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework region 4 of A-H.1 or A-H.2.

[0496] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a heavy chain variable domain comprising a framework region, e.g., framework region 3 (FR3), comprising a change, e.g., a substitution (e.g., a conservative substitution) at a position as described herein according to Kabat numbering. In some embodiments, FR3 comprises a Threonine at position 73, e.g., a substitution at position 73 according to Kabat numbering, e.g., a Glutamic Acid to Threonine substitution. In some embodiments, FR3 comprises a Glycine at position 94, e.g., a substitution at position 94 according to Kabat numbering, e.g., an Arginine to Glycine substitution. In some embodiments, the substitution is relative to a human germline heavy chain framework region sequence.WSGR Docket No.53676-769.601

[0497] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a heavy chain variable domain comprising a framework region 3 (FR3) comprising a Threonine at position 73, e.g., a substitution at position 73 according to Kabat numbering, e.g., a Glutamic Acid to Threonine substitution, and a Glycine at position 94, e.g., a substitution at position 94 according to Kabat numbering, e.g., a Arginine to Glycine substitution, e.g., as shown in the amino acid sequence of SEQ ID NO: 10.

[0498] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework regions 1-4 of A-H.1 or A-H.2, e.g., SEQ ID NO: 9. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework regions 1-4 of A-H.1, e.g., SEQ ID NO: 10. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the light chain framework regions 1-4 of A-H.2, e.g., SEQ ID NO: 11. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework regions 1-4 of A-H.1, e.g., SEQ ID NO: 9; and the light chain framework regions 1-4 of A-H.1, e.g., SEQ ID NO: 10. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises the heavy chain framework regions 1-4 of A-H.2, e.g., SEQ ID NO: 9; and the light chain framework regions 1-4 of A-H.2, e.g., SEQ ID NO: 11.

[0499] In some embodiments, the heavy or light chain variable domain, or both, of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes an amino acid sequence, which is substantially identical to an amino acid as described herein, e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical to a variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or as described in Table 1, or encoded by the nucleotide sequence in Table 1; or which differs at least 1 or 5 residues, but less than 40, 30, 20, or 10 residues, from a variable region of an antibody described herein.

[0500] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises at least one, two, three, or four antigen-binding regions, e.g., variable regions, having an amino acid sequence as set forth in Table 1, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, or which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the sequences shown in Table 1. In another embodiment, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes a VH and / or VL domain encoded by a nucleic acid having a nucleotide sequence as set forth in Table 1, or a sequence substantially identical thereto (e.g., a sequence at least about 85%, 90%, 95%, 99% or more identical thereto, or which differs by no more than 3, 6, 15, 30, or 45 nucleotides from the sequences shown in Table 1.

[0501] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 9, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acidWSGR Docket No.53676-769.601 sequence of SEQ ID NO: 9, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 9; and / or a VL domain comprising the amino acid sequence of SEQ ID NO: 10, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 10.

[0502] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises: a VH domain comprising the amino acid sequence of SEQ ID NO: 9, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 9; and / or a VL domain comprising the amino acid sequence of SEQ ID NO: 11, an amino acid sequence at least about 85%, 90%, 95%, 99% or more identical to the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence which differs by no more than 1, 2, 5, 10, or 15 amino acid residues from the amino acid sequence of SEQ ID NO: 11.

[0503] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule is a full antibody or fragment thereof (e.g., a Fab, F(ab')2, Fv, single domain antibody, or a single chain Fv fragment (scFv)). In embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule is a monoclonal antibody or an antibody with single specificity. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can also be a humanized, chimeric, camelid, shark, or an in vitro-generated antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, is a humanized antibody molecule. The heavy and light chains of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can be full-length (e.g., an antibody can include at least one, and preferably two, complete heavy chains, and at least one, and preferably two, complete light chains) or can include an antigen-binding fragment (e.g., a Fab, F(ab')2, Fv, a single chain Fv fragment, a single domain antibody, a diabody (dAb), a bivalent antibody, or bispecific antibody or fragment thereof, a single domain variant thereof, or a camelid antibody).

[0504] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, is in the form of a multispecific molecule, e.g., a bispecific molecule, e.g., as described herein.

[0505] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, has a heavy chain constant region (Fc) chosen from, e.g., the heavy chain constant regions of IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE. In some embodiments, the Fc region is chosen from the heavy chain constant regions of IgG1, IgG2, IgG3, and IgG4. In some embodiments, the Fc region is chosen from the heavy chain constant region of IgG1 or IgG2 (e.g., human IgG1, or IgG2). In some embodiments, the heavy chain constant region is human IgG1. In some embodiments, the Fc region comprises a Fc region variant, e.g., as described herein.WSGR Docket No.53676-769.601

[0506] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, has a light chain constant region chosen from, e.g., the light chain constant regions of kappa or lambda, preferably kappa (e.g., human kappa). In some embodiments, the constant region is altered, e.g., mutated, to modify the properties of the anti-TCRβV antibody molecule, e.g., anti- TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule (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). For example, the constant region is mutated at positions 296 (M to Y), 298 (S to T), 300 (T to E), 477 (H to K) and 478 (N to F) to alter Fc receptor binding (e.g., the mutated positions correspond to positions 132 (M to Y), 134 (S to T), 136 (T to E), 313 (H to K) and 314 (N to F) of SEQ ID NOs: 212 or 214; or positions 135 (M to Y), 137 (S to T), 139 (T to E), 316 (H to K) and 317 (N to F) of SEQ ID NOs: 215, 216, 217 or 218), e.g., relative to human IgG1.

[0507] Antibody A-H.1 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3278 and a light chain comprising the amino acid sequence of SEQ ID NO: 72. Antibody A-H.2 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 3278 and a light chain comprising the amino acid sequence of SEQ ID NO: 3279. Antibody A-H.68 comprises the amino acid sequence of SEQ ID NO: 1337, or a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto. Antibody A-H.69 comprises the amino acid sequence of SEQ ID NO: 1500, or a sequence having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto.

[0508] Additional exemplary humanized anti-TCRB V6 antibodies are provided in Table 1. In some embodiments, the anti-TCRβ V6 is antibody A, e.g., humanized antibody A (antibody A-H), as provided in Table 1. In some embodiments, the anti-TCRβV antibody comprises one or more (e.g., all three) of a LC CDR1, LC CDR2, and LC CDR3 provided in Table 1; and / or one or more (e.g., all three) of a HC CDR1, HC CDR2, and HC CDR3 provided in Table 1, or a sequence with at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto. In some embodiments, antibody A comprises a variable heavy chain (VH) and / or a variable light chain (VL) provided in Table 1, or a sequence with at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9%, or 100% sequence identity thereto.

[0509] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a VH of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A- H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A-H.19, A- H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A-H.31, A- H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A-H.43, A- H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-H.55, A- H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A-H.67, A- H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A-H.79, A- H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.WSGR Docket No.53676-769.601

[0510] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a VL of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A- H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A-H.19, A- H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A-H.31, A- H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A-H.43, A- H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-H.55, A- H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A-H.67, A- H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A-H.79, A- H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0511] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a VH of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A- H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A-H.19, A- H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A-H.31, A- H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A-H.43, A- H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A-H.55, A- H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A-H.67, A- H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A-H.79, A- H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto; and a VL of A-H.1, A-H.2, A-H.3, A-H.4, A-H.5, A-H.6, A-H.7, A-H.8, A-H.9, A-H.10, A-H.11, A-H.12, A-H.13, A-H.14, A-H.15, A-H.16, A-H.17, A-H.18, A- H.19, A-H.20, A-H.21, A-H.22, A-H.23, A-H.24, A-H.25, A-H.26, A-H.27, A-H.28, A-H.29, A-H.30, A- H.31, A-H.32, A-H.33, A-H.34, A-H.35, A-H.36, A-H.37, A-H.38, A-H.39, A-H.40, A-H.1, A-H.42, A- H.43, A-H.44, A-H.45, A-H.46, A-H.47, A-H.48, A-H.49, A-H.50, A-H.51, A-H.52, A-H.53, A-H.54, A- H.55, A-H.56, A-H.57, A-H.58, A-H.59, A-H.60, A-H.61, A-H.62, A-H.63, A-H.64, A-H.65, A-H.66, A- H.67, A-H.68, A-H.69, A-H.70, A-H.71, A-H.72, A-H.73, A-H.74, A-H.75, A-H.76, A-H.77, A-H.78, A- H.79, A-H.80, A-H.81, A-H.82, A-H.83, A-H.84, or A-H.85, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0512] Exemplary anti-TCRβV antibody molecules and the corresponding TCRβV subfamilies recognized by said anti-TCRβV antibody molecules are disclosed in Table 10A.

[0513] The various TCRβV subfamilies and / or subfamily members can be expressed at different levels in individuals, e.g., healthy individuals, as disclosed in Kitaura K. et al (2016), BMC Immunology vol 17: 38, the entire contents of which are hereby incorporated by reference. For example, TCRβ V6-5 is represented in approximately 3-6% healthy donors.

[0514] The representation of various TCRBV subfamilies and / or subfamily members can also be different in cancer cells. For example, TCRβV is present in about 3-6% of tumor infiltrating T cells irrespective of tumor type (see Li B. et al., Nature Genetics, 2016, vol:48(7):725-32 the entire contents ofWSGR Docket No.53676-769.601 which are hereby incorporated by references). Li et al., also disclose that TCRβ V6-5 is present at a high frequency in tumor cells. Anti-TCRβ V6 antibodies

[0515] In one aspect, provided herein is an anti-TCRβV antibody molecule that binds to human TCRβ V6, e.g., a TCRβ V6 subfamily comprising: 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 the TCRβ V6 subfamily comprises TCRβ V6-5*01 or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-4*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-9*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-8*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-5*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*02, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-6*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-2*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-3*01, or a variant thereof. In some embodiments, TCRβ V6 comprises TCRβ V6-1*01, or a variant thereof.

[0516] In some embodiments, TCRβ V6-5*01 is encoded by the nucleic acid sequence of SEQ ID NO: 43, or a sequence having 85%, 90%, 95%, 99% or more identity thereof. In some embodiments, TCRβ V6-5*01 comprises the amino acid sequence of SEQ ID NO: 44, or an amino acid sequence having 85%, 90%, 95%, 99% or more identity thereof.

[0517] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, is a non-murine antibody molecule, e.g., a human or humanized antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti- TCRβ V6-5*01) antibody molecule is a human antibody molecule. In some embodiments, the anti- TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule is a humanized antibody molecule.

[0518] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, is isolated or recombinant.

[0519] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises at least one antigen-binding region, e.g., a variable region or an antigen-binding fragment thereof, from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0520] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises at least one, two, three or four variable regions from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A- H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequenceWSGR Docket No.53676-769.601 substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0521] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody molecule described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0522] In some embodiments, the anti-TCRβV antibody molecule comprises a heavy chain variable region (VH) having a consensus sequence of SEQ ID NO: 231 or 3290.

[0523] SEQ ID NO: 231 - Consensus VH QVQLVQSGAEVKKPGSSVKVSCKASGH / T / G / YD / T / SFH / R / D / K / TL / D / K / T / NW / F / T / I / Y / GYIHWV RQAPGQGLEWMGR / WV / I / FF / S / YA / PGSGN / ST / V / Y / IK / RYNEKFKGRVTITADTSTSTAYMELSS LRSEDTAVYYCAG / VSY / IYSY / AD / GVLDYWGQGTTVTVSS

[0524] SEQ ID NO: 3290 - Consensus VH QVQLVQSGAEVKKPGSSVKVSCKASGX1X2FX3X4X5YIHWVRQAPGQGLEWMGX6X7X8X9G SGX10X11X12YNEKFKGRVTITADTSTSTAYMELSSLRSEDTAVYYCAX13SX14YSX15X16VLD YWGQGTTVTVSS, where-in: X1 is H or T or G or Y; X2 is D or T or S; X3 is H or R or D or K or T; X4 is L or D or K or T or N; X5 is W or F or T or I or Y or G; X6 is R or W; X7 is V or I or F; X8 is F or S or Y; X9 is A or P; X10 is N or S; X11 is T or V or Y or I; X12 is K or R; X13 is G or V; X14 is Y or I; X15 is Y or A; and X16 is D or G.

[0525] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A- H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0526] In some embodiments, the anti-TCRβV antibody molecule comprises a light chain variable region (VL) having a consensus sequence of SEQ ID NO: 230 or 3289.

[0527] SEQ ID NO: 230 - Consensus VL DIQMTQSPSFLSASVGDRVTITCKASQNVG / E / A / DN / DR / KVAWY / HQQKPGKAPKALIYSSSHRY K / SGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQFKSYPLTFGQGTKLEIK

[0528] SEQ ID NO: 3289 - Consensus VL DIQMTQSPSFLSASVGDRVTITCKASQNVX1X2X3VAWX4QQKPGKAPKALIYSSSHRYX5GVPS RFSGSGSGTEFTLTISSLQPEDFATYFCQQFKSYPLTFGQGTKLEIK, wherein X1 is G, E, A or D; X2 is N or D; X3 is R or K; X4 is Y or H; and X5 is K or S

[0529] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, comprises a heavy chain constant region for an IgG4, e.g., a human IgG4.WSGR Docket No.53676-769.601 In still another embodiment, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes a heavy chain constant region for an IgG1, e.g., a human IgG1. In some embodiments, the heavy chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0530] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes a kappa light chain constant region, e.g., a human kappa light chain constant region. In some embodiments, the light chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0531] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, or three complementarity determining regions (CDRs) from a heavy chain variable region (VH) of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0532] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a heavy chain variable region comprising an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.

[0533] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, or three complementarity determining regions (CDRs) from a light chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0534] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a light chain variable region comprising an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.

[0535] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, three, four, five or six CDRs (or collectively allWSGR Docket No.53676-769.601 of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 1, or encoded by a nucleotide sequence shown in Table 1.

[0536] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, molecule includes all six CDRs from an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1, or closely related CDRs, e.g., CDRs which are identical or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, may include any CDR described herein.

[0537] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 1) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A- H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 1.

[0538] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 1) from a light chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A- H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 1.

[0539] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, three, four, five, or six CDRs according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Kabat definition as set out in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of theWSGR Docket No.53676-769.601 aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Kabat et al. shown in Table 1.

[0540] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes all six CDRs according to Kabat et al. (e.g., all six CDRs according to the Kabat definition as set out in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A- H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Kabat et al. shown in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, may include any CDR described herein.

[0541] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody chosen from chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, e.g., the same canonical structures as at least loop 1 and / or loop 2 of the heavy and / or light chain variable domains of an antibody described herein. See, e.g., Chothia et al., (1992) J. Mol. Biol.227:799-817; Tomlinson et al., (1992) J. Mol. Biol.227:776-798 for descriptions of hypervariable loop canonical structures. These structures can be determined by inspection of the tables described in these references.

[0542] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 1) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or as described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 1.

[0543] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 1) from a light chain variable region of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A- H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three orWSGR Docket No.53676-769.601 four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 1.

[0544] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes at least one, two, three, four, five, or six CDRs according to Chothia et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Chothia definition as set out in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by the nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Chothia et al. shown in Table 1.

[0545] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, includes all six CDRs according to Chothia et al. (e.g., all six CDRs according to the Chothia definition as set out in Table 1) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody chosen from any one of A-H.1 to A-H.85, e.g., A-H.1, A-H.2 or A-H.68, or an antibody described in Table 1, or encoded by a nucleotide sequence in Table 1; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Chothia et al. shown in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, may include any CDR described herein.

[0546] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, molecule includes a combination of CDRs or hypervariable loops defined according to Kabat et al., Chothia et al., ImMunoGeneTics (IMGT) numbering system, or as described in Table 1.

[0547] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can contain any combination of CDRs or hypervariable loops according to the Kabat and Chothia definitions.

[0548] In some embodiments, a combined CDR as set out in Table 1 is a CDR that comprises a Kabat CDR and a Chothia CDR.

[0549] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, molecule includes a combination of CDRs or hypervariable loops identified as combined CDRs in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule, can contain any combination of CDRs or hypervariable loops according the “combined” CDRs are described in Table 1.WSGR Docket No.53676-769.601

[0550] In some embodiments, e.g., an embodiment comprising a variable region, a CDR (e.g., a combined CDR, Chothia CDR, Kabat CDR, or ImMunoGeneTics (IMGT) CDR), or other sequence referred to herein, e.g., in Table 1, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, a bivalent antibody molecule, a biparatopic antibody molecule, or an antibody molecule that comprises an antigen binding fragment of an antibody, e.g., a half antibody or antigen binding fragment of a half antibody. In certain embodiments the antibody molecule comprises a multispecific molecule, e.g., a bispecific molecule, e.g., as described herein.

[0551] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule includes: (i) one, two or all of a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3) of SEQ ID NO: 2, SEQ ID NO: 10 or SEQ ID NO: 11, and / or (ii) one, two or all of a heavy chain complementarity determining region 1 (HC CDR1), heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3) of SEQ ID NO: 1 or SEQ ID NO: 9.

[0552] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a LC CDR1, LC CDR2, and LC CDR3 of SEQ ID NO: 2, and a HC CDR1, HC CDR2, and HC CDR3 of SEQ ID NO: 1.

[0553] In some embodiments the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a LC CDR1, LC CDR2, and LC CDR3 of SEQ ID NO: 10, and a HC CDR1, HC CDR2, and HC CDR3 of SEQ ID NO: 9.

[0554] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a LC CDR1, LC CDR2, and LC CDR3 of SEQ ID NO: 11, and a HC CDR1, HC CDR2, and HC CDR3 of SEQ ID NO: 9.

[0555] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 6, a LC CDR2 amino acid sequence of SEQ ID NO: 7, or a LC CDR3 amino acid sequence of SEQ ID NO: 8; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 3, a HC CDR2 amino acid sequence of SEQ ID NO: 4, or a HC CDR3 amino acid sequence of SEQ ID NO: 5.

[0556] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 6, a LC CDR2 amino acid sequence of SEQ ID NO: 7, or a LC CDR3 amino acid sequence of SEQ ID NO: 8; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 3, a HC CDR2 amino acid sequence of SEQ ID NO: 4, or a HC CDR3 amino acid sequence of SEQ ID NO: 5.

[0557] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 51, a LC CDR2 amino acid sequence of SEQ ID NO: 52, or a LC CDR3 amino acid sequence of SEQ ID NO: 53;WSGR Docket No.53676-769.601 and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 45, a HC CDR2 amino acid sequence of SEQ ID NO: 46, or a HC CDR3 amino acid sequence of SEQ ID NO: 47.

[0558] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 51, a LC CDR2 amino acid sequence of SEQ ID NO: 52, or a LC CDR3 amino acid sequence of SEQ ID NO: 53; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 45, a HC CDR2 amino acid sequence of SEQ ID NO: 46, or a HC CDR3 amino acid sequence of SEQ ID NO: 47.

[0559] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 54, a LC CDR2 amino acid sequence of SEQ ID NO: 55, or a LC CDR3 amino acid sequence of SEQ ID NO: 56; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 48, a HC CDR2 amino acid sequence of SEQ ID NO: 49, or a HC CDR3 amino acid sequence of SEQ ID NO: 50.

[0560] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 54, a LC CDR2 amino acid sequence of SEQ ID NO: 55, or a LC CDR3 amino acid sequence of SEQ ID NO: 56; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 48, a HC CDR2 amino acid sequence of SEQ ID NO: 49, or a HC CDR3 amino acid sequence of SEQ ID NO: 50.

[0561] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a VH and / or a VL of an antibody described in Table 1, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0562] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V6 (e.g., anti-TCRβ V6-5*01) antibody molecule comprises a VH and a VL of an antibody described in Table 1, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.

[0563] In some embodiments, an anti-TCRVb antibody as described herein has an antigen binding domain having a VL having a consensus sequence of SEQ ID NO: 230, wherein position 30 is G, E, A or D; position 31 is N or D; position 32 is R or K; position 36 is Y or H; and / or position 56 is K or S.

[0564] In some embodiments, an anti-TCRVb antibody as described herein has an antigen binding domain having a VH having a consensus sequence of SEQ ID NO: 231, wherein: position 27 is H or T or G or Y; position 28 is D or T or S; position 30 is H or R or D or K or T; position 31 is L or D or K or T or N; position 32 is W or F or T or I or Y or G; position 49 is R or W; position 50 is V or I or F; position 51 is F or S or Y; position 52 is A or P; position 56 is N or S; position 57 is T or V or Y or I; position 58 is K or R; position 97 is G or V; position 99 is Y or I; position 102 is Y or A; and / or position 103 is D or G.

[0565] In some embodiments, the anti-TCRβ V6 antibody molecule comprises one or more sequences as described in Table 17 and Table 25, or a sequence with at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more identity thereto.WSGR Docket No.53676-769.601 Anti-TCRβ V12 antibodies

[0566] In one aspect, provided herein is an anti-TCRβV antibody molecule that binds to human TCRβ V12, e.g., a TCRβ V12 subfamily comprising: TCRβ V12-4*01, TCRβ V12-3*01 or TCRβ V12-5*01. In some embodiments the TCRβ V12 subfamily comprises TCRβ V12-4*01. In some embodiments the TCRβ V12 subfamily comprises TCRβ V12-3*01.

[0567] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, is a non-murine antibody molecule, e.g., a human or humanized antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule is a human antibody molecule. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule is a humanized antibody molecule.

[0568] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, is isolated or recombinant.

[0569] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, comprises at least one antigen-binding region, e.g., a variable region or an antigen-binding fragment thereof, from an antibody described herein, e.g., an antibody described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0570] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, comprises at least one, two, three or four variable regions from an antibody described herein, e.g., an antibody as described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0571] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, comprises at least one or two heavy chain variable regions from an antibody described herein, e.g., an antibody as described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0572] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, comprises at least one or two light chain variable regions from an antibody described herein, e.g., an antibody as described in Table 2, or encoded by a nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0573] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, comprises a heavy chain constant region for an IgG4, e.g., a human IgG4. In still another embodiment, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes a heavy chain constant region for an IgG1, e.g., a human IgG1. In some embodiments, the heavy chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.WSGR Docket No.53676-769.601

[0574] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes a kappa light chain constant region, e.g., a human kappa light chain constant region. In some embodiments, the light chain constant region comprises an amino sequence set forth in Table 3, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) thereto.

[0575] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes at least one, two, or three complementarity determining regions (CDRs) from a heavy chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0576] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a heavy chain variable region comprising an amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2.

[0577] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes at least one, two, or three complementarity determining regions (CDRs) from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences.

[0578] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes at least one, two, or three CDRs (or collectively all of the CDRs) from a light chain variable region comprising an amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2.

[0579] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, includes at least one, two, three, four, five or six CDRs (or collectively all of the CDRs) from a heavy and light chain variable region comprising an amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2. In some embodiments, one or more of the CDRs (or collectively all of the CDRs) have one, two, three, four, five, six or more changes, e.g., amino acid substitutions or deletions, relative to the amino acid sequence shown in Table 2, or encoded by a nucleotide sequence shown in Table 2.

[0580] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, molecule includes all six CDRs from an antibody described herein, e.g., an antibody asWSGR Docket No.53676-769.601 described in Table 2, or encoded by the nucleotide sequence in Table 2, or closely related CDRs, e.g., CDRs which are identical or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions). In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, may include any CDR described herein.

[0581] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 2) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 2.

[0582] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three CDRs according to Kabat et al. (e.g., at least one, two, or three CDRs according to the Kabat definition as set out in Table 2) from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Kabat et al. shown in Table 2.

[0583] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, three, four, five, or six CDRs according to Kabat et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Kabat definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Kabat et al. shown in Table 2.

[0584] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes all six CDRs according to Kabat et al. (e.g., all six CDRs according to the Kabat definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs accordingWSGR Docket No.53676-769.601 to Kabat et al. shown in Table 2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V12 antibody molecule may include any CDR described herein.

[0585] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three hypervariable loops that have the same canonical structures as the corresponding hypervariable loop of an antibody described herein, e.g., an antibody described in Table 2, e.g., the same canonical structures as at least loop 1 and / or loop 2 of the heavy and / or light chain variable domains of an antibody described herein. See, e.g., Chothia et al., (1992) J. Mol. Biol.227:799- 817; Tomlinson et al., (1992) J. Mol. Biol.227:776-798 for descriptions of hypervariable loop canonical structures. These structures can be determined by inspection of the tables described in these references.

[0586] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 2) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 2.

[0587] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three CDRs according to Chothia et al. (e.g., at least one, two, or three CDRs according to the Chothia definition as set out in Table 2) from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to Chothia et al. shown in Table 2.

[0588] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, three, four, five, or six CDRs according to Chothia et al. (e.g., at least one, two, three, four, five, or six CDRs according to the Chothia definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to Chothia et al. shown in Table 2.

[0589] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes all six CDRs according to Chothia et al. (e.g., all six CDRs according to the Chothia definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; orWSGR Docket No.53676-769.601 encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to Chothia et al. shown in Table 2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V12 antibody molecule may include any CDR described herein.

[0590] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three CDRs according to a combined CDR (e.g., at least one, two, or three CDRs according to the combined CDR definition as set out in Table 2) from a heavy chain variable region of an antibody described herein, e.g., an antibody chosen as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to combined CDR shown in Table 2.

[0591] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, or three CDRs according to a combined CDR (e.g., at least one, two, or three CDRs according to the combined CDR definition as set out in Table 2) from a light chain variable region of an antibody described herein, e.g., an antibody as described in Table 2, or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, or three CDRs according to a combined CDR shown in Table 2.

[0592] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes at least one, two, three, four, five, or six CDRs according to a combined CDR. (e.g., at least one, two, three, four, five, or six CDRs according to the combined CDR definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three or four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to one, two, three, four, five, or six CDRs according to a combined CDR shown in Table 2.

[0593] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes all six CDRs according to a combined CDR (e.g., all six CDRs according to the combined CDR definition as set out in Table 2) from the heavy and light chain variable regions of an antibody described herein, e.g., an antibody as described in Table 2, or encoded by the nucleotide sequence in Table 2; or encoded by the nucleotide sequence in Table 2; or a sequence substantially identical (e.g., at least 80%, 85%, 90%, 92%, 95%, 97%, 98%, 99% or higher identical) to any of the aforesaid sequences; or which have at least one amino acid alteration, but not more than two, three orWSGR Docket No.53676-769.601 four alterations (e.g., substitutions, deletions, or insertions, e.g., conservative substitutions) relative to all six CDRs according to a combined CDR shown in Table 2. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule may include any CDR described herein.

[0594] In some embodiments, a combined CDR as set out in Table 1 is a CDR that comprises a Kabat CDR and a Chothia CDR.

[0595] In some embodiments, the anti-TCRβV antibody molecule, e e.g., anti-TCRβ V12 antibody molecule, molecule includes a combination of CDRs or hypervariable loops identified as combined CDRs in Table 1. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, can contain any combination of CDRs or hypervariable loops according the “combined” CDRs are described in Table 1.

[0596] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes a combination of CDRs or hypervariable loops defined according to the Kabat et al. and Chothia et al., or as described in Table 1.

[0597] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule can contain any combination of CDRs or hypervariable loops according to the Kabat and Chothia definitions.

[0598] In some embodiments, e.g., an embodiment comprising a variable region, a CDR (e.g., a combined CDR, Chothia CDR, Kabat CDR, or ImMunoGeneTics (IMGT) CDR), or other sequence referred to herein, e.g., in Table 2, the antibody molecule is a monospecific antibody molecule, a bispecific antibody molecule, a bivalent antibody molecule, a biparatopic antibody molecule, or an antibody molecule that comprises an antigen binding fragment of an antibody, e.g., a half antibody or antigen binding fragment of a half antibody. In certain embodiments the antibody molecule comprises a multispecific molecule, e.g., a bispecific molecule, e.g., as described herein.

[0599] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes: (i) one, two or all of a light chain complementarity determining region 1 (LC CDR1), a light chain complementarity determining region 2 (LC CDR2), and a light chain complementarity determining region 3 (LC CDR3) of SEQ ID NO: 16, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29 or SEQ ID NO: 30, and / or (ii) one, two or all of a heavy chain complementarity determining region 1 (HC CDR1), heavy chain complementarity determining region 2 (HC CDR2), and a heavy chain complementarity determining region 3 (HC CDR3) of SEQ ID NO: 15, SEQ ID NO: 23, SEQ ID NO: 24 or SEQ ID NO: 25.

[0600] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 20, a LC CDR2 amino acid sequence of SEQ ID NO: 21, or a LC CDR3 amino acid sequence of SEQ ID NO: 22; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 17, a HC CDR2 amino acid sequence of SEQ ID NO: 18, or a HC CDR3 amino acid sequence of SEQ ID NO: 19.

[0601] In some embodiments, the anti-TCRβV antibody molecule,anti-TCRβ V12 antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequenceWSGR Docket No.53676-769.601 of SEQ ID NO: 20, a LC CDR2 amino acid sequence of SEQ ID NO: 21, and a LC CDR3 amino acid sequence of SEQ ID NO: 2; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 17, a HC CDR2 amino acid sequence of SEQ ID NO: 18, and a HC CDR3 amino acid sequence of SEQ ID NO: 19.

[0602] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 63, a LC CDR2 amino acid sequence of SEQ ID NO: 64, or a LC CDR3 amino acid sequence of SEQ ID NO: 65; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 57, a HC CDR2 amino acid sequence of SEQ ID NO: 58, or a HC CDR3 amino acid sequence of SEQ ID NO: 59.

[0603] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 63, a LC CDR2 amino acid sequence of SEQ ID NO: 64, or a LC CDR3 amino acid sequence of SEQ ID NO: 65; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 57, a HC CDR2 amino acid sequence of SEQ ID NO: 58, or a HC CDR3 amino acid sequence of SEQ ID NO: 59.

[0604] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises: (i) a LC CDR1 amino acid sequence of SEQ ID NO: 66, a LC CDR2 amino acid sequence of SEQ ID NO: 67, or a LC CDR3 amino acid sequence of SEQ ID NO: 68; and / or (ii) a HC CDR1 amino acid sequence of SEQ ID NO: 60, a HC CDR2 amino acid sequence of SEQ ID NO: 61, or a HC CDR3 amino acid sequence of SEQ ID NO: 62.

[0605] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises: (i) a light chain variable region (VL) comprising a LC CDR1 amino acid sequence of SEQ ID NO: 63, a LC CDR2 amino acid sequence of SEQ ID NO: 64, or a LC CDR3 amino acid sequence of SEQ ID NO: 65; and / or (ii) a heavy chain variable region (VH) comprising a HC CDR1 amino acid sequence of SEQ ID NO: 57, a HC CDR2 amino acid sequence of SEQ ID NO: 58, or a HC CDR3 amino acid sequence of SEQ ID NO: 59.

[0606] In some embodiments, the light or the heavy chain variable framework (e.g., the region encompassing at least FR1, FR2, FR3, and optionally FR4) of the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule can be chosen from: (a) a light or heavy chain variable framework including at least 80%, 85%, 87% 90%, 92%, 93%, 95%, 97%, 98%, or 100% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (b) a light or heavy chain variable framework including from 20% to 80%, 40% to 60%, 60% to 90%, or 70% to 95% of the amino acid residues from a human light or heavy chain variable framework, e.g., a light or heavy chain variable framework residue from a human mature antibody, a human germline sequence, or a human consensus sequence; (c) a non-human framework (e.g., a rodent framework); or (d) a non-human framework that has been modified, e.g., to remove antigenic or cytotoxic determinants, e.g., deimmunized, or partially humanized. In some embodiments, the light or heavy chain variableWSGR Docket No.53676-769.601 framework region (particularly FR1, FR2 and / or FR3) includes a light or heavy chain variable framework sequence at least 70, 75, 80, 85, 87, 88, 90, 92, 94, 95, 96, 97, 98, 99% identical or identical to the frameworks of a VL or VH segment of a human germline gene.

[0607] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule, comprises a heavy chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more changes, e.g., amino acid substitutions or deletions, from an amino acid sequence described in Table 2.e.g., the amino acid sequence of the FR region in the entire variable region, e.g., shown in FIGS.3A and 3B, or in SEQ ID NOs: 23-25.

[0608] Alternatively, or in combination with the heavy chain substitutions described herein the anti- TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain variable domain having at least one, two, three, four, five, six, seven, ten, fifteen, twenty or more amino acid changes, e.g., amino acid substitutions or deletions, from an amino acid sequence of an antibody described herein .e.g., the amino acid sequence of the FR region in the entire variable region, e.g., shown in FIGS.3A and 3B, or in SEQ ID NOs: 26-30.

[0609] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes one, two, three, or four heavy chain framework regions shown in FIG.3A, or a sequence substantially identical thereto.

[0610] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule includes one, two, three, or four light chain framework regions shown in FIG.3B, or a sequence substantially identical thereto. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework region 1 e.g., as shown in FIG. 3B. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework region 2 e.g., as shown in FIG.3B. In some embodiments, the anti- TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises the light chain framework region 3, e.g., as shown in FIG.3B. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti- TCRβ V12 antibody molecule comprises the light chain framework region 4, e.g., as shown in FIG.3B.

[0611] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a change, e.g., a substitution (e.g., a conservative substitution) at one or more, e.g., all, position as described herein according to Kabat numbering. In some embodiments, FR1 comprises an Aspartic Acid at position 1, e.g., a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution. In some embodiments, FR1 comprises an Asparagine at position 2, e.g., a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution. In some embodiments, FR1 comprises a Leucine at position 4, e.g., a substitution at position 4 according to Kabat numbering, e.g., a Methionine to Leucine substitution.

[0612] In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1),WSGR Docket No.53676-769.601 comprising a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution, a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution, and a substitution at position 4 according to Kabat numbering, e.g., a Methionine to Leucine substitution. In some embodiments, the anti-TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework region, e.g., framework region 1 (FR1), comprising a substitution at position 1 according to Kabat numbering, e.g., an Alanine to Aspartic Acid substitution, and a substitution at position 2 according to Kabat numbering, e.g., an Isoleucine to Asparagine substitution, Serine to Asparagine substitution or Tyrosine to Asparagine substitution. In some embodiments, the anti- TCRβV antibody molecule, e.g., anti-TCRβ V12 antibody molecule comprises a light chain comprising a framework...

Claims

WSGR Docket No.53676-769.601 CLAIMS WHAT IS CLAIMED IS:

1. A composition for treating a disease or condition in a subject in need thereof comprising: (a) a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region; and (b) a second agent comprising a second domain that binds to CD3.

2. A formulation for treating a disease or condition in a subject in need thereof comprising: (a) a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region; and (b) a second agent comprising a second domain that binds to CD3.

3. The composition of claim 1 or the formulation of claim 2, wherein the composition or the formulation comprises a therapeutically effective amount of the first agent and the second agent.

4. The composition of claim 1 or 3 or the formulation of claim 2 or 3, wherein the subject exhibits an improved therapeutic effect after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

5. The composition of any one of claims 1, 3, and 4, or the formulation of any one of claims 2-4, wherein the subject comprises more activated and / or expanded T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

6. The composition of any one of claims 1 and 3-5, or the formulation of any one of claims 2-5, wherein the subject comprises less exhausted T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

7. The composition of any one of claims 1 and 3-6, or the formulation of any one of claims 2-6, wherein the subject exhibits more anti-tumor efficacy after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

8. The composition of any one of claims 1 and 3-7, or the formulation of any one of claims 2-7, wherein the subject comprises T cells that are more cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

9. The composition of any one of claims 1 and 3-8, or the formulation of any one of claims 2-8, wherein the subject comprises more T cells that are cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

10. The composition of any one of claims 1 and 3-9, or the formulation of any one of claims 2-9, wherein the subject exhibits more cytotoxic activity against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

11. The composition of any one of claims 1 and 3-10, or the formulation of any one of claims 2-10, wherein the TCR variable region is a TCR β variable (TCRβV) region.

12. The composition of any one of claims 1 and 3-11, or the formulation of any one of claims 2-11, wherein the TCR variable region is a human TCR β variable (TCRβV) region.WSGR Docket No.53676-769.601 13. The composition or the formulation of claim 11 or 12, wherein the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of TCRβ V1 subfamily, TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 sub-family, TCRβ V6 subfamily, TCRβ V7 subfamily, TCRβ V8 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V14 sub-family, TCRβ V15 subfamily, TCRβ V16 subfamily, TCRβ V17 subfamily, TCRβ V18 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V22 subfamily TCRβ V23 subfamily, TCRβ V24 subfamily, TCRβ V25 subfamily, TCRβ V26 subfamily, TCRβ V27 subfamily, TCRβ V28 subfamily, TCRβ V29 subfamily, and TCRβ V30 subfamily.

14. The composition or the formulation of any one of claims 11-13, wherein the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V1 subfamily comprising TCRβ V1*01; (ii) TCRβ V2 subfamily comprising one or more selected from TCRβ V2*01, TCRβ V2*02, and TCRβ V2*03; (iii) TCRβ V3 subfamily comprising one or more selected from TCRβ V3-1*01 and TCRβ V3-1*02; (iv) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1*01, TCRβ V4-1*02, TCRβ V4-2*01, TCRβ V4-2*02, TCRβ V4-3*01, TCRβ V4-3*02, TCRβ V4-3*03, and TCRβ V4-3*04; (v) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-1*01, TCRβ V5-1*02, TCRβ V5-3*01, TCRβ V5-3*02, TCRβ V5-4*01, TCRβ V5-4*02, TCRβ V5-4*03, TCRβ V5-4*04, TCRβ V5-5*01, TCRβ V5-5*02, TCRβ V5-5*03, TCRβ V5-6*01, TCRβ V5-7*01, TCRβ V5- 8*01, and TCRβ V5-8*02; (vi) TCRβ V6 subfamily comprising one or more selected from TCRβ V6-1*01, TCRβ V6-2*01, TCRβ V6-3*01, TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-5*01, TCRβ V6-6*01, TCRβ V6-6*02, TCRβ V6-6*03, TCRβ V6-6*04, TCRβ V6-6*05, TCRβ V6-7*01, TCRβ V6-8*01, and TCRβ V6- 9*01; (vii) TCRβ V7 subfamily comprising one or more selected from TCRβ V7-1*01, TCRβ V7-2*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*04, TCRβ V7-3*01, TCRβ V7-3*02, TCRβ V7- 3*03, TCRβ V7-3*04, TCRβ V7-3*05, TCRβ V7-4*01, TCRβ V7-4*02, TCRβ V7-6*01, TCRβ V7-6*02, TCRβ V7-7*01, TCRβ V7-7*02, TCRβ V7-8*01, TCRβ V7-8*02, TCRβ V7-8*03, TCRβ V7-9*01, TCRβ V7-9*02, TCRβ V7-9*03, TCRβ V7-9*04, TCRβ V7-9*05, TCRβ V7- 9*06, and TCRβ V7-9*07. (viii) TCRβ V8 subfamily comprising one or more selected from TCRβ V8-1*01, TCRβ V8-1*02, TCRβ V8-2*01, and TCRβ V8-2*02; (ix) TCRβ V9 subfamily comprising one or more selected from TCRβ V9-1*01, TCRβ V9-1*02, and TCRβ V9-1*03; (x) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-1*03, TCRβ V10-2*01, TCRβ V10-2*02, TCRβ V10-3*01, TCRβ V10-3*02, TCRβ V10-3*03, and TCRβ V10-3*04;WSGR Docket No.53676-769.601 (xi) TCRβ V11 subfamily comprising TCRβ V11-1*01, TCRβ V11-2*01, TCRβ V11-2*02, TCRβ V11-2*03, TCRβ V11-3*01, TCRβ V11-3*02, TCRβ V11-3*03, and TCRβ V11-3*04; (xii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-3*01, TCRβ V12-4*01, TCRβ V12-4*02, and TCRβ V12-5*01; (xiii) TCRβ V13 subfamily comprising one or more selected from TCRβ V13*01 and TCRβ V13*02; (xiv) TCRβ V14 subfamily comprising one or more comprising from TCRβ V14*01 and TCRβ V14*02; (xv) TCRβ V15 subfamily comprising one or more selected from TCRβ V15*01, TCRβ V15*02, and TCRβ V15*03; (xvi) TCRβ V16 subfamily comprising one or more selected from TCRβ V16*01, TCRβ V16*02, and TCRβ V16*03; (xvii) TCRβ V17 subfamily comprising TCRβ V17*01; (xviii) TCRβ V18 subfamily comprising TCRβ V18*01; (xix) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01, TCRβ V19*02, and TCRβ V19*03; (xx) TCRβ V20 subfamily comprising one or more selected from TCRβ V20-1*01, TCRβ V20-1*02, TCRβ V20-1*03, TCRβ V20-1*04, TCRβ V20-1*05, TCRβ V20-1*06, and TCRβ V20-1*07; (xxi) TCRβ V21 subfamily comprising one or more selected from TCRβ V21-1*01 and TCRβ V21- 1*02; (xxii) TCRβ V22 subfamily comprising TCRβ V22-1*01; (xxiii) TCRβ V23 subfamily comprising TCRβ V23-1*01; (xxiv) TCRβ V24 subfamily comprising TCRβ V24-1*01; (xxv) TCRβ V25 subfamily comprising TCRβ V25-1*01; (xxvi) TCRβ V26 subfamily comprising TCRβ V26-1*01; (xxvii) TCRβ V27 subfamily comprising TCRβ V27*01; (xxviii) TCRβ V28 subfamily comprising TCRβ V28*01; (xxix) TCRβ V29 subfamily comprising one or more selected from TCRβ V29-1*01, TCRβ V29-1*02, and TCRβ V29-1*03; and (xxx) TCRβ V30 subfamily comprising one or more selected from TCRβ V30*01, TCRβ V30*02, TCRβ V30*03, TCRβ V30*04, and TCRβ V30*05.

15. The composition or the formulation of any one of claims 11-14, wherein the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRβV region.

16. The composition or the formulation of claim 15, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).WSGR Docket No.53676-769.601 17. The composition of any one of claims 1 and 3-10, or the formulation of any one of claims 2-10, wherein the TCR variable region is a TCR α variable region.

18. The composition of any one of claims 1, 3-10, and 17, or the formulation of any one of claims 2-10 and 17, wherein the TCR variable region is a human TCR α variable region.

19. The composition or the formulation of claim 17 or 18, wherein the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

20. The composition or the formulation of any one of claims 17-19, wherein the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02; (ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02; (iii) TCRα V3 subfamily comprising TCRα V3*01; (iv) TCRα V4 subfamily comprising TCRα V4*01; (v) TCRα V5 subfamily comprising TCRα V5*01 (vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06; (vii) TCRα V7 subfamily comprising TCRα V7*01; (viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8- 4*01, TCRα V8-4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8-6*02, and TCRα V8-7*01; (ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; (x) TCRα V10 subfamily comprising TCRα V10*01; (xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02; (xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02;WSGR Docket No.53676-769.601 (xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04; (xiv) TCRα V16 subfamily comprising TCRα V16*01; (xv) TCRα V17 subfamily comprising TCRα V17*01; (xvi) TCRα V18 subfamily comprising TCRα V18*01; (xvii) TCRα V19 subfamily comprising TCRα V19*01 (xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04; (xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02; (xx) TCRα V22 subfamily comprising TCRα V22*01; (xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04; (xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02; (xxiii) TCRα V25 subfamily comprising TCRα V25*01; (xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRα V26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02; (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01; (xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02; (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38-1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01; (xxxiii) TCRα V40 subfamily comprising TCRα V40*01; and (xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

21. The composition or the formulation of any one of claims 17-20, wherein the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRαV region.

22. The composition or the formulation of claim 21, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).WSGR Docket No.53676-769.601 23. The composition of any one of claims 1 and 3-22, or the formulation of any one of claims 2-22, wherein the first agent comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or fragment thereof, IgG2 heavy chain constant region or fragment thereof, IgG3 heavy chain constant region or fragment thereof, IgGA1 heavy chain constant region or fragment thereof, IgGA2 heavy chain constant region or fragment thereof, IgG4 heavy chain constant region or fragment thereof, IgJ heavy chain constant region or fragment thereof, IgM heavy chain constant region or fragment thereof, IgD heavy chain constant region or fragment thereof, and IgE heavy chain constant region or fragment thereof.

24. The composition of any one of claims 1 and 3-23, or the formulation of any one of claims 2-23, wherein the first agent comprises a kappa light chain constant region or fragment thereof, a lambda light chain constant region or fragment thereof, or a combination thereof.

25. The composition of any one of claims 1 and 3-24, or the formulation of any one of claims 2-24, wherein the first agent is a multifunctional molecule.

26. The composition of any one of claims 1 and 3-25, or the formulation of any one of claims 2-25, wherein the first agent comprises at least two non-contiguous polypeptide chains; wherein a first polypeptide chain of the at least two non-contiguous polypeptide chains comprises a first Fc region, and a second polypeptide chain of the at least two non-contiguous polypeptide chains comprises a second Fc region.

27. The composition or the formulation of claim 26, wherein the first Fc region and the second Fc region comprise an Fc interface with a knob-in-a hole.

28. The composition or the formulation of claim 26 or 27, wherein the first Fc region, the second Fc region, or a combination thereof comprises an Asn297Ala mutation, a Leu234Ala / Leu235Ala mutation, or a combination thereof.

29. The composition of any one of claims 1 and 3-28, or the formulation of any one of claims 2-28, wherein the first agent is a multispecific molecule that further comprises one or more of a tumor- targeting moiety, a cytokine molecule, a stromal modifying moiety, or an immune cell engager.

30. The composition of any one of claims 1 and 3-29, or the formulation of any one of claims 2-29, wherein the first agent is a multispecific molecule that further comprises a cytokine molecule.

31. The composition of any one of claims 1 and 3-30, or the formulation of any one of claims 2-30, wherein the first agent comprises the following configuration: A-, B-[dimerization module]-C, -D, wherein: (a) the dimerization module comprises a first immunoglobulin chain constant region and a second immunoglobulin chain constant region; (b) A and C are linked to the first immunoglobulin chain constant region; and B and D are linked to the second immunoglobulin chain constant region; (c) A, B, C, and D are independently (i) absent; (ii) the first binder; (iii) the tumor targeting moiety, (iv) the cytokine molecule; (v) stromal modifying moiety; or (vi) the immune cell engager; andWSGR Docket No.53676-769.601 wherein at least one, two, or three of A, B, C, and D comprises the first agent.

32. The composition or the formulation of claim 31, wherein the first immunoglobulin chain constant region comprises the first fragment crystallizable region (Fc region) and the second first immunoglobulin chain constant region comprises the second fragment crystallizable region (Fc region).

33. The composition or the formulation of claim 32, wherein dimerization of the first Fc region and the second Fc region is enhanced by providing an Fc interface of the first Fc region and the second Fc region with one or more of a paired cavity-protuberance, an electrostatic interaction, or a strand- exchange, such that a greater ratio of heteromultimer to homomultimer forms relative to a non- engineered interface.

34. The composition or the formulation of any one of claims 29-33, 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, a macrophage cell engager, and any combination thereof.

35. The composition or the formulation of any one of claims 29-34, wherein the cytokine molecule is selected from the group consisting of inter-leukin-2 (IL-2) or functional variant thereof, interleukin- 7 (IL-7) or functional variant thereof, in-terleukin-12 (IL-12) or functional variant thereof, interleukin-15 (IL-15) or functional variant thereof, interleukin-18 (IL-18) or functional variant thereof, interleukin-21 (IL-21) or functional variant thereof, interferon gamma or functional variant thereof, and any combination thereof.

36. The composition or the formulation of any one of claims 29-35, wherein the cytokine molecule comprises interleukin-2 (IL-2) or functional variant thereof.

37. The composition or the formulation of claim 36, wherein the interleukin-2 (IL-2) or functional variant thereof comprises the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

38. The composition or the formulation of any one of claims 29-35, wherein the cytokine molecule comprises interleukin-15 (IL-15) or functional variant thereof.

39. The composition or the formulation of claim 38, wherein the interleukin-15 (IL-15) or functional variant thereof comprises the sequence of SEQ ID NO: 2170.

40. The composition or the formulation of claim 38 or 39, wherein the cytokine molecule further comprises an IL15Ralpha dimerizing domain covalently linked the interleukin-15 (IL-15) or functional variant thereof.

41. The composition or the formulation of claim 40, wherein the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or functional variant thereof.

42. The composition or the formulation of claim 40 or 41, wherein the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

43. The composition or the formulation of any one of claims 40-42, wherein the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.WSGR Docket No.53676-769.601 44. The composition or the formulation of claim 43, wherein the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

45. The composition or the formulation of any one of claims 40-44, wherein the cytokine molecule comprises the sequence of SEQ ID NO: 3474.

46. The composition of any one of claims 1 and 3-45, or the formulation of any one of claims 2-45, wherein the second domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to CD3.

47. The composition or the formulation of claim 46, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

48. The composition of any one of claims 1 and 3-47, or the formulation of any one of claims 2-47, wherein the second domain binds to CD3ε.

49. The composition of any one of claims 1 and 3-48, or the formulation of any one of claims 2-48, wherein the second domain binds to human CD3ε.

50. The composition or the formulation of claim 48 or 49, wherein the second domain is selected from the group consisting of muromonab-CD3, an antigen binding domain thereof, or a functional fragment or variant thereof, otelixizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, teplizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, and visilizumab, an antigen binding domain thereof, or a functional fragment or variant thereof.

51. The composition of any one of claims 1 and 3-50, or the formulation of any one of claims 2-50, wherein the second agent further comprises a full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen.

52. The composition or the formulation of claim 51, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen is operatively linked to the second domain.

53. The composition or the formulation of claim 51 or 52, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, TRP-1 / -2, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, MUC-1, CA-125, BAGE, GAGE, NY-ESO-1, β- catenin, CDK4, CDC27, α actinin-4, TRP1 / gp75, TRP2, gp100, Melan-A / MART1, gangliosides,WSGR Docket No.53676-769.601 WT1, EphA3, Epidermal growth factor receptor (EGFR), MART-2, MART-1, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

54. The composition of any one of claims 1 and 3-53, or the formulation of any one of claims 2-53, wherein the condition or disease is cancer.

55. The composition or the formulation of claim 54, wherein the cancer is a solid tumor, a hematological cancer, a metastatic cancer, a soft tissue tumor, or a combination thereof.

56. The composition or the formulation of claim 55, wherein the cancer is the solid tumor, and wherein the solid tumor is selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

57. The composition or the formulation of claim 55, wherein the cancer is the hematological cancer, and wherein the hematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T-cell lymphoma, acute lymphocytic leukemia, and a combination thereof.

58. The composition or the formulation of claim 57, wherein the Non-Hodgkin’s lymphoma is selected from the group consisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

59. The composition or the formulation of claim 58, wherein the T-cell lymphoma is peripheral T-cell lymphoma.

60. The composition or the formulation of any one of claims 54-59, wherein the cancer is characterized by a cancer antigen present on the cancer.

61. The composition or the formulation of claim 60, wherein the cancer antigen is a tumor antigen, a stromal antigen, or a hematological antigen.

62. The composition or the formulation of claim 60 or 61, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, ganglioside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, TRP1 / gp75, TRP2, gangliosides, WT1, Epi-dermal growth factor receptor (EGFR), MART-2, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG,WSGR Docket No.53676-769.601 TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Integrins, Carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

63. The composition of any one of claims 1 and 3-62, wherein the composition is pharmaceutical composition.

64. The formulation of any one of claims 2-62, wherein the formulation is pharmaceutical formulation.

65. The composition of any one of claims 1 and 3-63, or the formulation of any one of claims 2-62 and 64, wherein the composition or formulation further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

66. A method of treating a disease or condition in a subject in need thereof comprising: (a) administering a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region; and (b) administering a second agent comprising a second domain that binds to CD3; wherein a therapeutically effective amount of the first agent and the second agent is administered to the subject, thereby treating the disease or condition in the subject.

67. A method of treating a disease or condition in a subject in need thereof comprising administering to the subject a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, wherein the subject has been previously treated with a second agent comprising a second domain that binds to CD3, thereby treating the disease or condition in the subject.

68. A method of improving a therapeutic efficacy of a second agent comprising a second domain that binds to CD3 in a subject in need thereof comprising: administering a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, thereby improving the therapeutic efficacy of the second agent in the subject.

69. A combination therapy for treating a disease or condition in a subject in need thereof a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, and a second agent comprising a second domain that binds to CD3.

70. The method of any one of claims 66-68 or the combination therapy of claim 69, wherein administering the first agent and the second agent induces an improved therapeutic effect in the subject, relative to a subject administered the second agent without the first agent.

71. The method of any one of claims 66-68 and 70 or the combination therapy of claim 69 or 70, wherein administering the first agent and the second agent induces more activated and / or expanded T cells in the subject, relative to a subject administered the second agent without the first agent.

72. The method of any one of claims 66-68, 70, and 71 or the combination therapy of any one of claims 69-71, wherein administering the first agent and the second agent induces less exhausted T cells in the subject, relative to a subject administered the second agent without the first agent.

73. The method of any one of claims 66-68 and 70-72 or the combination therapy of any one of claims 69-72, wherein administering the first agent and the second agent induces more anti-tumor efficacy in the subject, relative to a subject administered the second agent without the first agent.WSGR Docket No.53676-769.601 74. The method of any one of claims 66-68 and 70-73 or the combination therapy of any one of claims 69-73, wherein administering the first agent and the second agent induces more cytotoxicity against tumor cells or cancer cells in the subject, relative to a subject administered the second agent without the first agent.

75. The method of any one of claims 66-68 and 70-74 or the combination therapy of any one of claims 69-74, wherein administering the first agent and the second agent expands or activates more T cells that are cytotoxic against tumor cells or cancer cells in the subject, relative to a subject administered the second agent without the first agent.

76. The method of any one of claims 66-68 and 70-75 or the combination therapy of any one of claims 69-75, wherein administering the first agent and the second agent induces more cytotoxic activity against tumor cells or cancer cells in the subject, relative to a subject administered the second agent without the first agent.

77. The method of any one of claims 66-68 and 70-76 or the combination therapy of any one of claims 69-76, wherein the first agent and the second agent are not concomitantly administered to the subject.

78. The method of any one of claims 66-68 and 70-76 or the combination therapy of any one of claims 69-76, wherein the first agent and the second agent are concomitantly administered to the subject.

79. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years after the subject has been treated with a dose of the second agent.

80. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years after the subject has been treated with a first dose of the second agent.

81. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years after the subject has been treated with a last dose of the second agent.

82. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with the second agent.WSGR Docket No.53676-769.601 83. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with a dose of the second agent.

84. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with a first dose of the second agent.

85. The method of any one of claims 66-68 and 70-78 or the combination therapy of any one of claims 69-78, wherein the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years after being treated with a last dose of the second agent.

86. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 hour to 24 hours after being treated with a dose of the second agent.

87. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 day to 7 days after being treated with a dose of the second agent.

88. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 week to 4 weeks after being treated with a dose of the second agent.

89. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 month to 12 months after being treated with a dose of the second agent.

90. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 year to 3 years after being treated with a dose of the second agent.

91. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 hour to 24 hours after being treated with a first dose of the second agent.

92. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 day to 7 days after being treated with a first dose of the second agent.WSGR Docket No.53676-769.601 93. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 week to 4 weeks after being treated with a first dose of the second agent.

94. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 month to 12 months after being treated with a first dose of the second agent.

95. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 year to 3 years after being treated with a first dose of the second agent.

96. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 hour to 24 hours after being treated with a last dose of the second agent.

97. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 week to 4 weeks after being treated with a last dose of the second agent.

98. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 month to 12 months after being treated with a last dose of the second agent.

99. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject from 1 year to 3 years after being treated with a last dose of the second agent.

100. The method of any one of claims 66-68 and 70-85 or the combination therapy of any one of claims 69-85, wherein the method comprises administering the first agent to the subject on the same day the subject had been previously treated with the second agent.

101. The method of any one of claims 66-68 and 70-100 or the combination therapy of any one of claims 69-100, wherein the method is repeated twice or more.

102. The method of any one of claims 66-68 and 70-101 or the combination therapy of any one of claims 69-101, wherein the administration of the first agent comprises one or more doses.

103. The method of any one of claims 66-68 and 70-102 or the combination therapy of any one of claims 69-102, wherein the administration of the first agent comprises two or more doses.

104. The method of any one of claims 66-68 and 70-103 or the combination therapy of any one of claims 69-103, wherein the administration of the first agent comprises one dose per week.

105. The method of any one of claims 66-68 and 70-103 or the combination therapy of any one of claims 69-103, wherein the administration of the first agent comprises two or more doses per week.

106. The method of any one of claims 66-68 and 70-103 or the combination therapy of any one of claims 69-103, wherein the first agent is administered to the subject once every two weeks.

107. The method of any one of claims 66-68 and 70-103 or the combination therapy of any one of claims 69-103, wherein the first agent is administered to the subject once every three weeks.WSGR Docket No.53676-769.601 108. The method of any one of claims 66-68 and 70-104 or the combination therapy of any one of claims 69-104, wherein the first agent is administered to the subject once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

109. The method of any one of claims 66-68, 70-103, and 105 or the combination therapy of any one of claims 69-103 and 105, wherein the administration of the first agent comprises two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

110. The method of any one of claims 66-68, 70-103, and 106 or the combination therapy of any one of claims 69-103 and 106, wherein the first agent is administered to the subject once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

111. The method of any one of claims 66-68, 70-103, and 107 or the combination therapy of any one of claims 69-103 and 107, wherein the first agent is administered to the subject once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

112. The method of any one of claims 66-68 and 70-111 or the combination therapy of any one of claims 69-111, wherein the administration of the second agent comprises one or more doses.

113. The method of any one of claims 66-68 and 70-112 or the combination therapy of any one of claims 69-112, wherein the administration of the second agent comprises two or more doses.

114. The method of any one of claims 66-68 and 70-113 or the combination therapy of any one of claims 69-113, wherein the administration of the second agent comprises one dose per week.

115. The method of any one of claims 66-68 and 70-113 or the combination therapy of any one of claims 69-113, wherein the administration of the second agent comprises two or more doses per week.

116. The method of any one of claims 66-68 and 70-113 or the combination therapy of any one of claims 69-113, wherein the second agent is administered to the subject once every two weeks.

117. The method of any one of claims 66-68 and 70-113 or the combination therapy of any one of claims 69-113, wherein the second agent is administered to the subject once every three weeks.

118. The method of any one of claims 66-68 and 70-114 or the combination therapy of any one of claims 69-114, wherein the second agent is administered to the subject once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

119. The method of any one of claims 66-68, 70-113, and 115 or the combination therapy of any one of claims 69-113 and 115, wherein the administration of the second agent comprises two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

120. The method of any one of claims 66-68, 70-113, and 116 or the combination therapy of any one of claims 69-113 and 116, wherein the second agent is administered to the subject once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.WSGR Docket No.53676-769.601 121. The method of any one of claims 66-68, 70-113, and 117 or the combination therapy of any one of claims 69-113 and 117, wherein the second agent is administered to the subject once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

122. The method of any one of claims 66-68 and 70-121 or the combination therapy of any one of claims 69-121, wherein the method comprises administering the first agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years prior to administering the second agent to the subject.

123. The method of any one of claims 66-68 and 70-122 or the combination therapy of any one of claims 69-122, wherein the method comprises administering the first agent to the subject at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years prior to administering the second agent to the subject.

124. The method of any one of claims 66-68 and 70-123 or the combination therapy of any one of claims 69-123, wherein the method comprises administering the first agent to the subject from 1 hour to 24 hours prior to administering the second agent to the subject.

125. The method of any one of claims 66-68 and 70-123 or the combination therapy of any one of claims 69-123, wherein the method comprises administering the first agent to the subject from 1 day to 7 days prior to administering the second agent to the subject.

126. The method of any one of claims 66-68 and 70-123 or the combination therapy of any one of claims 69-123, wherein the method comprises administering the first agent to the subject from 1 week to 4 weeks prior to administering the second agent to the subject.

127. The method of any one of claims 66-68 and 70-123 or the combination therapy of any one of claims 69-123, wherein the method comprises administering the first agent to the subject from 1 month to 12 months prior to administering the second agent to the subject.

128. The method of any one of claims 66-68 and 70-123 or the combination therapy of any one of claims 69-123, wherein the method comprises administering the first agent to the subject from 1 year to 3 years prior to administering the second agent to the subject.

129. The method of any one of claims 66-68 and 70-128 or the combination therapy of any one of claims 69-128, wherein the administration of the first agent comprises two or more doses, and each dose comprises a same amount of the first agent.

130. The method of any one of claims 66-68 and 70-128 or the combination therapy of any one of claims 69-128, wherein the administration of the first agent comprises two or more doses, and each dose comprises different amounts of the first agent.

131. The method of any one of claims 66-68 and 70-130 or the combination therapy of any one of claims 69-130 wherein the administration of the second agent comprises two or more doses, and each dose comprises a same amount of the second agent.WSGR Docket No.53676-769.601 132. The method of any one of claims 66-68 and 70-130 or the combination therapy of any one of claims 69-130, wherein the administration of the second agent comprises two or more doses, and each dose comprises different amounts of the second agent.

133. The method of any one of claims 66-68 and 70-132 or the combination therapy of any one of claims 69-132, wherein the first agent is administered to the subject at a dose of from about 0.001 mg / kg to about 20 mg / kg.

134. The method of any one of claims 66-68 and 70-133 or the combination therapy of any one of claims 69-133, wherein the second agent is administered to the subject at a dose of from about 0.001 mg / kg to about 20 mg / kg.

135. The method of any one of claims 66-68 and 70-134 or the combination therapy of any one of claims 69-134, wherein the method comprises administering the first agent and the second agent to the subject simultaneously or on the same day.

136. The method of any one of claims 66-68 and 70-135 or the combination therapy of any one of claims 69-135, wherein the disease or condition in the subject is treated to a higher extent relative to a corresponding method in which the first agent is administered and the second agent is not administered.

137. The method of any one of claims 66-68 and 70-136 or the combination therapy of any one of claims 69-136, wherein the method is more effective to treat the disease or condition in the subject relative to a method in which the first agent is administered to the subject but not the second agent.

138. The method of any one of claims 66-68 and 70-137 or the combination therapy of any one of claims 69-137, wherein the method is more effective to treat the disease or condition in the subject relative to a method in which the second agent is administered to the subject but not the first agent.

139. The method of any one of claims 66-68 and 70-138 or the combination therapy of any one of claims 69-138, wherein the TCR variable region is a TCR β variable (TCRβV) region.

140. The method of any one of claims 66-68 and 70-139 or the combination therapy of any one of claims 69-139, wherein the TCR variable region is a human TCR β variable (TCRβV) region.

141. The method or the combination therapy of claim 139 or 140, wherein the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of TCRβ V1 subfamily, TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 sub-family, TCRβ V6 subfamily, TCRβ V7 subfamily, TCRβ V8 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V14 sub-family, TCRβ V15 subfamily, TCRβ V16 subfamily, TCRβ V17 subfamily, TCRβ V18 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V22 subfamily TCRβ V23 subfamily, TCRβ V24 subfamily, TCRβ V25 subfamily, TCRβ V26 subfamily, TCRβ V27 subfamily, TCRβ V28 subfamily, TCRβ V29 subfamily, and TCRβ V30 subfamily.

142. The method or the combination therapy of any one of claims 139-141, wherein the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V1 subfamily comprising TCRβ V1*01;WSGR Docket No.53676-769.601 (ii) TCRβ V2 subfamily comprising one or more selected from TCRβ V2*01, TCRβ V2*02, and TCRβ V2*03; (iii) TCRβ V3 subfamily comprising one or more selected from TCRβ V3-1*01 and TCRβ V3-1*02; (iv) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1*01, TCRβ V4-1*02, TCRβ V4-2*01, TCRβ V4-2*02, TCRβ V4-3*01, TCRβ V4-3*02, TCRβ V4-3*03, and TCRβ V4-3*04; (v) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-1*01, TCRβ V5-1*02, TCRβ V5-3*01, TCRβ V5-3*02, TCRβ V5-4*01, TCRβ V5-4*02, TCRβ V5-4*03, TCRβ V5-4*04, TCRβ V5-5*01, TCRβ V5-5*02, TCRβ V5-5*03, TCRβ V5-6*01, TCRβ V5-7*01, TCRβ V5- 8*01, and TCRβ V5-8*02; (vi) TCRβ V6 subfamily comprising one or more selected from TCRβ V6-1*01, TCRβ V6-2*01, TCRβ V6-3*01, TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-5*01, TCRβ V6-6*01, TCRβ V6-6*02, TCRβ V6-6*03, TCRβ V6-6*04, TCRβ V6-6*05, TCRβ V6-7*01, TCRβ V6-8*01, and TCRβ V6- 9*01; (vii) TCRβ V7 subfamily comprising one or more selected from TCRβ V7-1*01, TCRβ V7-2*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*04, TCRβ V7-3*01, TCRβ V7-3*02, TCRβ V7- 3*03, TCRβ V7-3*04, TCRβ V7-3*05, TCRβ V7-4*01, TCRβ V7-4*02, TCRβ V7-6*01, TCRβ V7-6*02, TCRβ V7-7*01, TCRβ V7-7*02, TCRβ V7-8*01, TCRβ V7-8*02, TCRβ V7-8*03, TCRβ V7-9*01, TCRβ V7-9*02, TCRβ V7-9*03, TCRβ V7-9*04, TCRβ V7-9*05, TCRβ V7- 9*06, and TCRβ V7-9*07. (viii) TCRβ V8 subfamily comprising one or more selected from TCRβ V8-1*01, TCRβ V8-1*02, TCRβ V8-2*01, and TCRβ V8-2*02; (ix) TCRβ V9 subfamily comprising one or more selected from TCRβ V9-1*01, TCRβ V9-1*02, and TCRβ V9-1*03; (x) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-1*03, TCRβ V10-2*01, TCRβ V10-2*02, TCRβ V10-3*01, TCRβ V10-3*02, TCRβ V10-3*03, and TCRβ V10-3*04; (xi) TCRβ V11 subfamily comprising TCRβ V11-1*01, TCRβ V11-2*01, TCRβ V11-2*02, TCRβ V11-2*03, TCRβ V11-3*01, TCRβ V11-3*02, TCRβ V11-3*03, and TCRβ V11-3*04; (xii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-3*01, TCRβ V12-4*01, TCRβ V12-4*02, and TCRβ V12-5*01; (xiii) TCRβ V13 subfamily comprising one or more selected from TCRβ V13*01 and TCRβ V13*02; (xiv) TCRβ V14 subfamily comprising one or more comprising from TCRβ V14*01 and TCRβ V14*02; (xv) TCRβ V15 subfamily comprising one or more selected from TCRβ V15*01, TCRβ V15*02, and TCRβ V15*03; (xvi) TCRβ V16 subfamily comprising one or more selected from TCRβ V16*01, TCRβ V16*02, and TCRβ V16*03; (xvii) TCRβ V17 subfamily comprising TCRβ V17*01;WSGR Docket No.53676-769.601 (xviii) TCRβ V18 subfamily comprising TCRβ V18*01; (xix) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01, TCRβ V19*02, and TCRβ V19*03; (xx) TCRβ V20 subfamily comprising one or more selected from TCRβ V20-1*01, TCRβ V20-1*02, TCRβ V20-1*03, TCRβ V20-1*04, TCRβ V20-1*05, TCRβ V20-1*06, and TCRβ V20-1*07; (xxi) TCRβ V21 subfamily comprising one or more selected from TCRβ V21-1*01 and TCRβ V21- 1*02; (xxii) TCRβ V22 subfamily comprising TCRβ V22-1*01; (xxiii) TCRβ V23 subfamily comprising TCRβ V23-1*01; (xxiv) TCRβ V24 subfamily comprising TCRβ V24-1*01; (xxv) TCRβ V25 subfamily comprising TCRβ V25-1*01; (xxvi) TCRβ V26 subfamily comprising TCRβ V26-1*01; (xxvii) TCRβ V27 subfamily comprising TCRβ V27*01; (xxviii) TCRβ V28 subfamily comprising TCRβ V28*01; (xxix) TCRβ V29 subfamily comprising one or more selected from TCRβ V29-1*01, TCRβ V29-1*02, and TCRβ V29-1*03; and (xxx) TCRβ V30 subfamily comprising one or more selected from TCRβ V30*01, TCRβ V30*02, TCRβ V30*03, TCRβ V30*04, and TCRβ V30*05.

143. The method or the combination therapy of any one of claims 139-142, wherein the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRβV region.

144. The method or the combination therapy of claim 143, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

145. The method of any one of claims 66-68 and 70-137 or the combination therapy of any one of claims 69-138, wherein the TCR variable region is a TCR α variable region.

146. The method of any one of claims 66-68, 70-137, and 145 or the combination therapy of any one of claims 69-138 and 145, wherein the TCR variable region is a human TCR α variable region.

147. The method or the combination therapy of claim 145 or 146, wherein the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36WSGR Docket No.53676-769.601 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

148. The method or the combination therapy of any one of claims 145-147, wherein the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02; (ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02; (iii) TCRα V3 subfamily comprising TCRα V3*01; (iv) TCRα V4 subfamily comprising TCRα V4*01; (v) TCRα V5 subfamily comprising TCRα V5*01 (vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06; (vii) TCRα V7 subfamily comprising TCRα V7*01; (viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8- 4*01, TCRα V8-4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8-6*02, and TCRα V8-7*01; (ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; (x) TCRα V10 subfamily comprising TCRα V10*01; (xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02; (xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02; (xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04; (xiv) TCRα V16 subfamily comprising TCRα V16*01; (xv) TCRα V17 subfamily comprising TCRα V17*01; (xvi) TCRα V18 subfamily comprising TCRα V18*01; (xvii) TCRα V19 subfamily comprising TCRα V19*01 (xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04; (xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02; (xx) TCRα V22 subfamily comprising TCRα V22*01; (xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04; (xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02; (xxiii) TCRα V25 subfamily comprising TCRα V25*01;WSGR Docket No.53676-769.601 (xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRα V26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02; (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01; (xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02; (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38-1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01; (xxxiii) TCRα V40 subfamily comprising TCRα V40*01; and (xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

149. The method or the combination therapy of any one of claims 145-148, wherein the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRαV region.

150. The method or the combination therapy of claim 149, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

151. The method of any one of claims 66-68 and 70-150 or the combination therapy of any one of claims 69-150, wherein the first agent comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or fragment thereof, IgG2 heavy chain constant region or fragment thereof, IgG3 heavy chain constant region or fragment thereof, IgGA1 heavy chain constant region or fragment thereof, IgGA2 heavy chain constant region or fragment thereof, IgG4 heavy chain constant region or fragment thereof, IgJ heavy chain constant region or fragment thereof, IgM heavy chain constant region or fragment thereof, IgD heavy chain constant region or fragment thereof, and IgE heavy chain constant region or fragment thereof.

152. The method of any one of claims 66-68 and 70-151 or the combination therapy of any one of claims 69-151, wherein the first agent comprises a kappa light chain constant region or fragment thereof, a lambda light chain constant region or fragment thereof, or a combination thereof.

153. The method of any one of claims 66-68 and 70-152 or the combination therapy of any one of claims 69-152, wherein the first agent is a multifunctional molecule.

154. The method of any one of claims 66-68 and 70-153 or the combination therapy of any one of claims 69-153, wherein the first agent comprises at least two non-contiguous polypeptide chains;WSGR Docket No.53676-769.601 wherein a first polypeptide chain of the at least two non-contiguous polypeptide chains comprises a first Fc region, and a second polypeptide chain of the at least two non-contiguous polypeptide chains comprises a second Fc region.

155. The method or the combination therapy of claim 154, wherein the first Fc region and the second Fc region comprise an Fc interface with a knob-in-a hole.

156. The method or the combination therapy of claim 154 or 155, wherein the first Fc region, the second Fc region, or a combination thereof comprises an Asn297Ala mutation, a Leu234Ala / Leu235Ala mutation, or a combination thereof.

157. The method of any one of claims 66-68 and 70-156 or the combination therapy of any one of claims 69-156, wherein the first agent is a multispecific molecule that further comprises one or more of a tumor-targeting moiety, a cytokine molecule, a stromal modifying moiety, or an immune cell engager.

158. The method of any one of claims 66-68 and 70-157 or the combination therapy of any one of claims 69-157, wherein the first agent is a multispecific molecule that further comprises a cytokine molecule.

159. The method of any one of claims 66-68 and 70-158 or the combination therapy of any one of claims 69-158, wherein the first agent comprises the following configuration: A-, B-[dimerization module]-C, -D, wherein: (a) the dimerization module comprises a first immunoglobulin chain constant region and a second immunoglobulin chain constant region; (b) A and C are linked to the first immunoglobulin chain constant region; and B and D are linked to the second immunoglobulin chain constant region; (c) A, B, C, and D are independently (i) absent; (ii) the first binder; (iii) the tumor targeting moiety, (iv) the cytokine molecule; (v) stromal modifying moiety; or (vi) the immune cell engager; and wherein at least one, two, or three of A, B, C, and D comprises the first agent.

160. The method or the combination therapy of claim 159, wherein the first immunoglobulin chain constant region comprises the first fragment crystallizable region (Fc region) and the second first immunoglobulin chain constant region comprises the second fragment crystallizable region (Fc region).

161. The method or the combination therapy of claim 160, wherein dimerization of the first Fc region and the second Fc region is enhanced by providing an Fc interface of the first Fc region and the second Fc region with one or more of a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms relative to a non-engineered interface.

162. The method or the combination therapy of any one of claims 157-161, 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, a macrophage cell engager, and any combination thereof.WSGR Docket No.53676-769.601 163. The method or the combination therapy of any one of claims 157-162, wherein the cytokine molecule is selected from the group consisting of inter-leukin-2 (IL-2) or functional variant thereof, interleukin-7 (IL-7) or functional variant thereof, in-terleukin-12 (IL-12) or functional variant thereof, interleukin-15 (IL-15) or functional variant thereof, interleukin-18 (IL-18) or functional variant thereof, interleukin-21 (IL-21) or functional variant thereof, interferon gamma or functional variant thereof, and any combination thereof.

164. The method or the combination therapy of any one of claims 157-163, wherein the cytokine molecule comprises interleukin-2 (IL-2) or functional variant thereof.

165. The method or the combination therapy of claim 164, wherein the interleukin-2 (IL-2) or functional variant thereof comprises the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

166. The method or the combination therapy of any one of claims 157-163, wherein the cytokine molecule comprises interleukin-15 (IL-15) or functional variant thereof.

167. The method or the combination therapy of claim 166, wherein the interleukin-15 (IL-15) or functional variant thereof comprises the sequence of SEQ ID NO: 2170.

168. The method or the combination therapy of claim 166 or 167, wherein the cytokine molecule further comprises an IL15Ralpha dimerizing domain covalently linked the interleukin-15 (IL-15) or functional variant thereof.

169. The method or the combination therapy of claim 168, wherein the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or functional variant thereof.

170. The method or the combination therapy of claim 168 or 169, wherein the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

171. The method or the combination therapy of any one of claims 168-170, wherein the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

172. The method or the combination therapy of claim 171, wherein the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

173. The method or the combination therapy of any one of claims 168-172, wherein the cytokine molecule comprises the sequence of SEQ ID NO: 3474.

174. The method of any one of claims 66-68 and 70-173 or the combination therapy of any one of claims 69-173, wherein the second domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to CD3.

175. The method or the combination therapy of claim 174, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).WSGR Docket No.53676-769.601 176. The method of any one of claims 66-68 and 70-175 or the combination therapy of any one of claims 69-175, wherein the second domain binds to CD3ε.

177. The method of any one of claims 66-68 and 70-176 or the combination therapy of any one of claims 69-176, wherein the second domain binds to human CD3ε.

178. The method or the combination therapy of claim 176 or 177, wherein the second domain is selected from the group consisting of muromonab-CD3, an antigen binding domain thereof, or a functional fragment or variant thereof, otelixizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, teplizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, and visilizumab, an antigen binding domain thereof, or a functional fragment or variant thereof.

179. The method of any one of claims 66-68 and 70-178 or the combination therapy of any one of claims 69-178, wherein the second agent further comprises a full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen.

180. The method or the combination therapy of claim 179, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen is operatively linked to the second domain.

181. The method or the combination therapy of claim 179 or 180, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate- specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan- A / MART-1, Gp100 / pmel17, Tyrosinase, TRP-1 / -2, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, MUC-1, CA-125, BAGE, GAGE, NY- ESO-1, β-catenin, CDK4, CDC27, α actinin-4, TRP1 / gp75, TRP2, gp100, Melan-A / MART1, gangliosides, WT1, EphA3, Epidermal growth factor receptor (EGFR), MART-2, MART-1, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

182. The method of any one of claims 66-68 and 70-181 or the combination therapy of any one of claims 69-181, wherein the condition or disease is cancer.

183. The method or the combination therapy of claim 182, wherein the cancer is a solid tumor, a hematological cancer, a metastatic cancer, a soft tissue tumor, or a combination thereof.

184. The method or the combination therapy of claim 183, wherein the cancer is the solid tumor, and wherein the solid tumor is selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.WSGR Docket No.53676-769.601 185. The method or the combination therapy of claim 183, wherein the cancer is the hematological cancer, and wherein the hematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T-cell lymphoma, acute lymphocytic leukemia, and a combination thereof.

186. The method or the combination therapy of claim 185, wherein the Non-Hodgkin’s lymphoma is selected from the group consisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

187. The method or the combination therapy of claim 186, wherein the T-cell lymphoma is peripheral T- cell lymphoma.

188. The method or the combination therapy of any one of claims 182-187, wherein the cancer is characterized by a cancer antigen present on the cancer.

189. The method or the combination therapy of claim 188, wherein the cancer antigen is a tumor antigen, a stromal antigen, or a hematological antigen.

190. The method or the combination therapy of claim 188 or 189, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, ganglioside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate- specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan- A / MART-1, Gp100 / pmel17, Tyrosinase, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, TRP1 / gp75, TRP2, gangliosides, WT1, Epi-dermal growth factor receptor (EGFR), MART-2, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Integrins, Carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

191. The method of any one of claims 66-68 and 70-190 or the combination therapy of any one of claims 69-190, wherein the first agent is administered to the subject prior to, concurrently, or after administration of the second agent to the subject.

192. A kit comprising: (i) a first agent comprising a first domain that binds to a TCR (T Cell Receptor) variable region, (ii) a second agent comprising a second domain that binds to CD3, and (iii) instructions for administering the first agent and the second agent to a human subject in need thereof to treat a disease or condition in the human subject.WSGR Docket No.53676-769.601 193. The kit of claim 192, wherein the instructions comprise instructions for administering a therapeutic effective amount of the first agent to the human subject.

194. The kit of claim 192 or 193, wherein the instructions comprise instructions for administering a therapeutic effective amount of the second agent to the human subject.

195. The kit of any one of claims 192-194, wherein the instructions comprise instructions for frequency of administering the first agent to the human subject.

196. The kit of any one of claims 192-195, wherein the instructions comprise instructions for frequency of administering the second agent to the human subject.

197. The kit of any one of claims 192-196, wherein the instructions comprise instructions to administer the second agent to the human subject prior to administering the first agent to the human subject.

198. The kit of any one of claims 192-196, wherein the instructions comprise instructions to administer the second agent and the first agent not concomitantly to the human subject.

199. The kit of any one of claims 192-198, wherein the instructions comprise instructions to administer to the human subject one or more doses of the second agent.

200. The kit of any one of claims 192-199, wherein the instructions comprise instructions to administer to the human subject two or more doses of the second agent.

201. The kit of any one of claims 192-200, wherein the instructions comprise instructions to administer to the human subject the second agent one dose per week.

202. The kit of any one of claims 192-200, wherein the instructions comprise instructions to administer to the human subject the second agent two or more doses per week.

203. The kit of any one of claims 192-200, wherein the instructions comprise instructions to administer to the human subject the second agent once every two weeks.

204. The kit of any one of claims 192-200, wherein the instructions comprise instructions to administer to the human subject the second agent once every three weeks.

205. The kit of any one of claims 192-201, wherein the instructions comprise instructions to administer to the human subject the second agent once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

206. The kit of any one of claims 192-200 and 202, wherein the instructions comprise instructions to administer to the human subject the second agent two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

207. The kit of any one of claims 192-200 and 203, wherein the instructions comprise instructions to administer to the human subject the second agent once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

208. The kit of any one of claims 192-200 and 204, wherein the instructions comprise instructions to administer to the human subject the second agent once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

209. The kit of any one of claims 192-208, wherein the instructions comprise instructions to administer one or more doses of the first agent.WSGR Docket No.53676-769.601 210. The kit of any one of claims 192-209, wherein the instructions comprise instructions to administer two or more doses of the first agent.

211. The kit of any one of claims 192-210, wherein the administration of the first agent comprises one dose per week.

212. The kit of any one of claims 192-210, wherein the administration of the first agent comprises two or more doses per week.

213. The kit of any one of claims 192-210, wherein the instructions comprise instructions to administer to the human subject the first agent once every two weeks.

214. The kit of any one of claims 192-210, wherein the instructions comprise instructions to administer to the human subject the first agent once every three weeks.

215. The kit of any one of claims 192-211, wherein the instructions comprise instructions to administer to the human subject the first agent once every week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

216. The kit of any one of claims 192-210 and 212, wherein the instructions comprise instructions to administer to the human subject the first agent two or more doses per week for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

217. The kit of any one of claims 192-210 and 213, wherein the instructions comprise instructions to administer to the human subject the first agent once every two weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

218. The kit of any one of claims 192-210 and 214, wherein the instructions comprise instructions to administer to the human subject the first agent once every three weeks for at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years.

219. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent to the subject at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at least 1, 2, 3, 4, 5, 6, or 7 days, at least 1, 2, 3, or 4 weeks, at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at least 1, 2, or 3 years prior to administering the first agent to the subject.

220. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 24 hours, at most 1, 2, 3, 4, 5, 6, or 7 days, at most 1, 2, 3, or 4 weeks, at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 months, or at most 1, 2, or 3 years prior to administering the first agent to the subject.

221. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent from 1 hour to 24 hours prior to administering the first agent to the subject.

222. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent from 1 day to 7 days prior to administering the first agent to the subject.WSGR Docket No.53676-769.601 223. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent from 1 week to 4 weeks prior to administering the first agent to the subject.

224. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent to the subject from 1 month to 12 months prior to administering the first agent to the subject.

225. The kit of any one of claims 192-218, wherein the instructions comprise instructions to administer to the human subject the second agent to the subject from 1 year to 3 years prior to administering the first agent to the subject.

226. The kit of any one of claims 192-225, wherein the instructions comprise instructions to administer to the human subject two or more doses of the second agent, and each dose comprises a same amount of the second agent.

227. The kit of any one of claims 192-225, wherein the instructions comprise instructions to administer to the human subject two or more doses of the second agent, and each dose comprises different amounts of the second agent.

228. The kit of any one of claims 192-227, wherein the instructions comprise instructions to administer to the human subject two or more doses of the first agent, and each dose comprises a same amount of the first agent.

229. The kit of any one of claims 192-227, wherein the instructions comprise instructions to administer to the human subject two or more doses of the first agent, and each dose comprises different amounts of the first agent.

230. The kit of any one of claims 192-229, wherein the instructions comprise instructions to administer to the human subject the second agent at a dose of from about 0.001 mg / kg to about 20 mg / kg.

231. The kit of any one of claims 192-230, wherein the instructions comprise instructions to administer to the human subject the first agent at a dose of from about 0.001 mg / kg to about 20 mg / kg.

232. The kit of any one of claims 192-231, wherein the instructions comprise instructions to administer to the human subject the second agent and the first agent to the subject simultaneously or on the same day.

233. The kit of any one of claims 192-232, wherein the instructions comprise instructions to administer the first agent to the subject prior to, concurrently, or after administration of the second agent to the subject.

234. The kit of any one of claims 192-233, wherein the subject exhibits an improved therapeutic effect after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

235. The kit of any one of claims 192-234, wherein the subject comprises more activated and / or expanded T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.WSGR Docket No.53676-769.601 236. The kit of any one of claims 192-235, wherein the subject comprises less exhausted T cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

237. The kit of any one of claims 192-236, wherein the subject exhibits more anti-tumor efficacy after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

238. The kit of any one of claims 192-237, wherein the subject comprises T cells that are more cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

239. The kit of any one of claims 192-238, wherein the subject comprises more T cells that are cytotoxic against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

240. The kit of any one of claims 192-239, wherein the subject exhibits more cytotoxic activity against tumor cells or cancer cells after administration of the first agent and the second agent, relative to a subject administered the second agent without the first agent.

241. The kit of any one of claims 192-240, wherein the TCR variable region is a TCR β variable (TCRβV) region.

242. The kit of any one of claims 192-241, wherein the TCR variable region is a human TCR β variable (TCRβV) region.

243. The kit of claim 241 or 242, wherein the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of TCRβ V1 subfamily, TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 sub-family, TCRβ V6 subfamily, TCRβ V7 subfamily, TCRβ V8 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V14 sub-family, TCRβ V15 subfamily, TCRβ V16 subfamily, TCRβ V17 subfamily, TCRβ V18 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V22 subfamily TCRβ V23 subfamily, TCRβ V24 subfamily, TCRβ V25 subfamily, TCRβ V26 subfamily, TCRβ V27 subfamily, TCRβ V28 subfamily, TCRβ V29 subfamily, and TCRβ V30 subfamily.

244. The kit of any one of claims 241-243, wherein the first domain binds to one or more of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V1 subfamily comprising TCRβ V1*01; (ii) TCRβ V2 subfamily comprising one or more selected from TCRβ V2*01, TCRβ V2*02, and TCRβ V2*03; (iii) TCRβ V3 subfamily comprising one or more selected from TCRβ V3-1*01 and TCRβ V3-1*02; (iv) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1*01, TCRβ V4-1*02, TCRβ V4-2*01, TCRβ V4-2*02, TCRβ V4-3*01, TCRβ V4-3*02, TCRβ V4-3*03, and TCRβ V4-3*04; (v) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-1*01, TCRβ V5-1*02, TCRβ V5-3*01, TCRβ V5-3*02, TCRβ V5-4*01, TCRβ V5-4*02, TCRβ V5-4*03, TCRβ V5-4*04,WSGR Docket No.53676-769.601 TCRβ V5-5*01, TCRβ V5-5*02, TCRβ V5-5*03, TCRβ V5-6*01, TCRβ V5-7*01, TCRβ V5- 8*01, and TCRβ V5-8*02; (vi) TCRβ V6 subfamily comprising one or more selected from TCRβ V6-1*01, TCRβ V6-2*01, TCRβ V6-3*01, TCRβ V6-4*01, TCRβ V6-4*02, TCRβ V6-5*01, TCRβ V6-6*01, TCRβ V6-6*02, TCRβ V6-6*03, TCRβ V6-6*04, TCRβ V6-6*05, TCRβ V6-7*01, TCRβ V6-8*01, and TCRβ V6- 9*01; (vii) TCRβ V7 subfamily comprising one or more selected from TCRβ V7-1*01, TCRβ V7-2*01, TCRβ V7-2*02, TCRβ V7-2*03, TCRβ V7-2*04, TCRβ V7-3*01, TCRβ V7-3*02, TCRβ V7- 3*03, TCRβ V7-3*04, TCRβ V7-3*05, TCRβ V7-4*01, TCRβ V7-4*02, TCRβ V7-6*01, TCRβ V7-6*02, TCRβ V7-7*01, TCRβ V7-7*02, TCRβ V7-8*01, TCRβ V7-8*02, TCRβ V7-8*03, TCRβ V7-9*01, TCRβ V7-9*02, TCRβ V7-9*03, TCRβ V7-9*04, TCRβ V7-9*05, TCRβ V7- 9*06, and TCRβ V7-9*07. (viii) TCRβ V8 subfamily comprising one or more selected from TCRβ V8-1*01, TCRβ V8-1*02, TCRβ V8-2*01, and TCRβ V8-2*02; (ix) TCRβ V9 subfamily comprising one or more selected from TCRβ V9-1*01, TCRβ V9-1*02, and TCRβ V9-1*03; (x) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10-1*02, TCRβ V10-1*03, TCRβ V10-2*01, TCRβ V10-2*02, TCRβ V10-3*01, TCRβ V10-3*02, TCRβ V10-3*03, and TCRβ V10-3*04; (xi) TCRβ V11 subfamily comprising TCRβ V11-1*01, TCRβ V11-2*01, TCRβ V11-2*02, TCRβ V11-2*03, TCRβ V11-3*01, TCRβ V11-3*02, TCRβ V11-3*03, and TCRβ V11-3*04; (xii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-3*01, TCRβ V12-4*01, TCRβ V12-4*02, and TCRβ V12-5*01; (xiii) TCRβ V13 subfamily comprising one or more selected from TCRβ V13*01 and TCRβ V13*02; (xiv) TCRβ V14 subfamily comprising one or more comprising from TCRβ V14*01 and TCRβ V14*02; (xv) TCRβ V15 subfamily comprising one or more selected from TCRβ V15*01, TCRβ V15*02, and TCRβ V15*03; (xvi) TCRβ V16 subfamily comprising one or more selected from TCRβ V16*01, TCRβ V16*02, and TCRβ V16*03; (xvii) TCRβ V17 subfamily comprising TCRβ V17*01; (xviii) TCRβ V18 subfamily comprising TCRβ V18*01; (xix) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01, TCRβ V19*02, and TCRβ V19*03; (xx) TCRβ V20 subfamily comprising one or more selected from TCRβ V20-1*01, TCRβ V20-1*02, TCRβ V20-1*03, TCRβ V20-1*04, TCRβ V20-1*05, TCRβ V20-1*06, and TCRβ V20-1*07; (xxi) TCRβ V21 subfamily comprising one or more selected from TCRβ V21-1*01 and TCRβ V21- 1*02;WSGR Docket No.53676-769.601 (xxii) TCRβ V22 subfamily comprising TCRβ V22-1*01; (xxiii) TCRβ V23 subfamily comprising TCRβ V23-1*01; (xxiv) TCRβ V24 subfamily comprising TCRβ V24-1*01; (xxv) TCRβ V25 subfamily comprising TCRβ V25-1*01; (xxvi) TCRβ V26 subfamily comprising TCRβ V26-1*01; (xxvii) TCRβ V27 subfamily comprising TCRβ V27*01; (xxviii) TCRβ V28 subfamily comprising TCRβ V28*01; (xxix) TCRβ V29 subfamily comprising one or more selected from TCRβ V29-1*01, TCRβ V29-1*02, and TCRβ V29-1*03; and (xxx) TCRβ V30 subfamily comprising one or more selected from TCRβ V30*01, TCRβ V30*02, TCRβ V30*03, TCRβ V30*04, and TCRβ V30*05.

245. The kit of any one of claims 241-244, wherein the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRβV region.

246. The kit of claim 245, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

247. The kit of any one of claims 192-240, wherein the TCR variable region is a TCR α variable region.

248. The kit of any one of claims 192-240 and 247, wherein the TCR variable region is a human TCR α variable region.

249. The kit of claim 247 or 248, wherein the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

250. The kit of any one of claims 247-249, wherein the first domain binds to one or more of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02; (ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02; (iii) TCRα V3 subfamily comprising TCRα V3*01; (iv) TCRα V4 subfamily comprising TCRα V4*01;WSGR Docket No.53676-769.601 (v) TCRα V5 subfamily comprising TCRα V5*01 (vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06; (vii) TCRα V7 subfamily comprising TCRα V7*01; (viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8- 4*01, TCRα V8-4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8-6*02, and TCRα V8-7*01; (ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; (x) TCRα V10 subfamily comprising TCRα V10*01; (xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02; (xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02; (xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04; (xiv) TCRα V16 subfamily comprising TCRα V16*01; (xv) TCRα V17 subfamily comprising TCRα V17*01; (xvi) TCRα V18 subfamily comprising TCRα V18*01; (xvii) TCRα V19 subfamily comprising TCRα V19*01 (xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04; (xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02; (xx) TCRα V22 subfamily comprising TCRα V22*01; (xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04; (xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02; (xxiii) TCRα V25 subfamily comprising TCRα V25*01; (xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRα V26-1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02; (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01; (xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02;WSGR Docket No.53676-769.601 (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38-1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01; (xxxiii) TCRα V40 subfamily comprising TCRα V40*01; and (xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

251. The kit of any one of claims 247-250, wherein the first domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRαV region.

252. The kit of claim 251, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

253. The kit of any one of claims 192-252, wherein the first agent comprises one or more heavy chain constant regions selected from the group consisting of IgG1 heavy chain constant region or fragment thereof, IgG2 heavy chain constant region or fragment thereof, IgG3 heavy chain constant region or fragment thereof, IgGA1 heavy chain constant region or fragment thereof, IgGA2 heavy chain constant region or fragment thereof, IgG4 heavy chain constant region or fragment thereof, IgJ heavy chain constant region or fragment thereof, IgM heavy chain constant region or fragment thereof, IgD heavy chain constant region or fragment thereof, and IgE heavy chain constant region or fragment thereof.

254. The kit of any one of claims 192-253, wherein the first agent comprises a kappa light chain constant region or fragment thereof, a lambda light chain constant region or fragment thereof, or a combination thereof.

255. The kit of any one of claims 192-254, wherein the first agent is a multifunctional molecule.

256. The kit of any one of claims 192-255, wherein the first agent comprises at least two non-contiguous polypeptide chains; wherein a first polypeptide chain of the at least two non-contiguous polypeptide chains comprises a first Fc region, and a second polypeptide chain of the at least two non-contiguous polypeptide chains comprises a second Fc region.

257. The kit of claim 256, wherein the first Fc region and the second Fc region comprise an Fc interface with a knob-in-a hole.

258. The kit of claim 256 or 257, wherein the first Fc region, the second Fc region, or a combination thereof comprises an Asn297Ala mutation, a Leu234Ala / Leu235Ala mutation, or a combination thereof.WSGR Docket No.53676-769.601 259. The kit of any one of claims 192-258, wherein the first agent is a multispecific molecule that further comprises one or more of a tumor-targeting moiety, a cytokine molecule, a stromal modifying moiety, or an immune cell engager.

260. The kit of any one of claims 192-259, wherein the first agent is a multispecific molecule that further comprises a cytokine molecule.

261. The kit of any one of claims 192-260, wherein the first agent comprises the following configuration: A-, B-[dimerization module]-C, -D, wherein: (a) the dimerization module comprises a first immunoglobulin chain constant region and a second immunoglobulin chain constant region; (b) A and C are linked to the first immunoglobulin chain constant region; and B and D are linked to the second immunoglobulin chain constant region; (c) A, B, C, and D are independently (i) absent; (ii) the first binder; (iii) the tumor targeting moiety, (iv) the cytokine molecule; (v) stromal modifying moiety; or (vi) the immune cell engager; and wherein at least one, two, or three of A, B, C, and D comprises the first agent.

262. The kit of claim 261, wherein the first immunoglobulin chain constant region comprises the first fragment crystallizable region (Fc region) and the second first immunoglobulin chain constant region comprises the second fragment crystallizable region (Fc region).

263. The kit of claim 262, wherein dimerization of the first Fc region and the second Fc region is enhanced by providing an Fc interface of the first Fc region and the second Fc region with one or more of a paired cavity-protuberance, an electrostatic interaction, or a strand-exchange, such that a greater ratio of heteromultimer to homomultimer forms relative to a non-engineered interface.

264. The kit of any one of claims 258-263, 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, a macrophage cell engager, and any combination thereof.

265. The kit of any one of claims 258-264, wherein the cytokine molecule is selected from the group consisting of inter-leukin-2 (IL-2) or functional variant thereof, interleukin-7 (IL-7) or functional variant thereof, in-terleukin-12 (IL-12) or functional variant thereof, interleukin-15 (IL-15) or functional variant thereof, interleukin-18 (IL-18) or functional variant thereof, interleukin-21 (IL- 21) or functional variant thereof, interferon gamma or functional variant thereof, and any combination thereof.

266. The kit of any one of claims 258-265, wherein the cytokine molecule comprises interleukin-2 (IL-2) or functional variant thereof.

267. The kit of claim 266, wherein the interleukin-2 (IL-2) or functional variant thereof comprises the sequence of SEQ ID NO: 2270 or SEQ ID NO: 2191.

268. The kit of any one of claims 258-265, wherein the cytokine molecule comprises interleukin-15 (IL- 15) or functional variant thereof.WSGR Docket No.53676-769.601 269. The kit of claim 268, wherein the interleukin-15 (IL-15) or functional variant thereof comprises the sequence of SEQ ID NO: 2170.

270. The kit of claim 268 or 269, wherein the cytokine molecule further comprises an IL15Ralpha dimerizing domain covalently linked the interleukin-15 (IL-15) or functional variant thereof.

271. The kit of claim 270, wherein the IL15Ralpha dimerizing domain comprises an IL-15 receptor alpha sushi domain or functional variant thereof.

272. The kit of claim 270 or 271, wherein the IL15Ralpha dimerizing domain comprises the sequence of SEQ ID NO: 3472.

273. The kit of any one of claims 270-272, wherein the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker.

274. The kit of claim 273, wherein the interleukin-15 (IL-15) or functional variant thereof is covalently linked to the IL15Ralpha dimerizing domain via a linker comprising the sequence of SEQ ID NO: 3473.

275. The kit of any one of claims 270-274, wherein the cytokine molecule comprises the sequence of SEQ ID NO: 3474.

276. The kit of any one of claims 192-275, wherein the second domain is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to CD3.

277. The kit of claim 276, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

278. The kit of any one of claims 192-277, wherein the second domain binds to CD3ε.

279. The kit of any one of claims 192-278, wherein the second domain binds to human CD3ε.

280. The kit of claim 278 or 279, wherein the second domain is selected from the group consisting of muromonab-CD3, an antigen binding domain thereof, or a functional fragment or variant thereof, otelixizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, teplizumab, an antigen binding domain thereof, or a functional fragment or variant thereof, and visilizumab, an antigen binding domain thereof, or a functional fragment or variant thereof.

281. The kit of any one of claims 192-280, wherein the second agent further comprises a full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen.

282. The kit of claim 281, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant that binds to a cancer antigen is operatively linked to the second domain.

283. The kit of claim 281 or 282, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING- 4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase,WSGR Docket No.53676-769.601 SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, TRP-1 / -2, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, MUC-1, CA-125, BAGE, GAGE, NY-ESO-1, β-catenin, CDK4, CDC27, α actinin-4, TRP1 / gp75, TRP2, gp100, Melan-A / MART1, gangliosides, WT1, EphA3, Epidermal growth factor receptor (EGFR), MART-2, MART-1, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

284. The kit of any one of claims 192-283, wherein the condition or disease is cancer.

285. The kit of claim 284, wherein the cancer is a solid tumor, a hematological cancer, a metastatic cancer, a soft tissue tumor, or a combination thereof.

286. The kit of claim 285, wherein the cancer is the solid tumor, and wherein the solid tumor is selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

287. The kit of claim 285, wherein the cancer is the hematological cancer, and wherein the hematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T-cell lymphoma, acute lymphocytic leukemia, and a combination thereof.

288. The kit of claim 287, wherein the Non-Hodgkin’s lymphoma is selected from the group consisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

289. The kit of claim 288, wherein the T-cell lymphoma is peripheral T-cell lymphoma.

290. The kit of any one of claims 284-289, wherein the cancer is characterized by a cancer antigen present on the cancer.

291. The kit of claim 290, wherein the cancer antigen is a tumor antigen, a stromal antigen, or a hematological antigen.

292. The kit of claim 290 or 291, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, ganglioside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING- 4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, CA-125, BAGE, GAGE, CDC27, α actinin-4, TRP1 / gp75, TRP2, gangliosides, WT1, Epi-dermal growth factor receptor (EGFR), MART-2, MUC1, MUC2, MUM1, MUM2, MUM3, MSLN, NA88-1, NPM, OA1, OGT, RCC, RU11, RU12, SAGE, TRG, TSTA, FolateWSGR Docket No.53676-769.601 receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Integrins, Carbohydrates, IGF1R, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, and tenascin W.

293. Use of the composition of any one of claims 1, 3-63, and 65, the formulation of any one of claims 2- 62, 64 and 65, or the kit of any one of claims 192-292 for the method of any one of claims 66-68 and 70-191 or the combination therapy of any one of claims 69-191.

294. A multifunctional molecule comprising: (a) a tumor-associated antigen binding moiety, wherein the tumor-associated antigen is DLL-3 or CLDN18.2; and (b) a TCR (T Cell Receptor) variable region (TCRV)-binding moiety.

295. The multifunctional molecule of claim 294, wherein the TCRV-binding moiety binds to a TCR β variable (TCRβV) region.

296. The multifunctional molecule of claim 294, wherein the TCRV-binding moiety binds to a TCR α variable (TCRαV) region.

297. The multifunctional molecule of any one of claims 294-296, further comprising at least one cytokine molecule or functional fragment or functional variant thereof.

298. The multifunctional molecule of claim 297, wherein the TCRV-binding moiety is covalently linked to the at least one cytokine molecule or functional fragment or functional variant thereof.

299. The multifunctional molecule of any one of claims 294-298, wherein the multifunctional molecule comprises a first polypeptide comprising a first portion of a dimerization module, and a second polypeptide comprising a second portion of the dimerization module; wherein the first polypeptide and the second polypeptide are non-contiguous, and wherein the tumor-associated antigen binding moiety is linked to the first portion of the dimerization module, and the at least one cytokine molecule or functional fragment or functional variant thereof is linked to the first portion of the dimerization module, the second portion of the dimerization module, or a combination thereof.

300. The multifunctional molecule of claim 299, wherein: (i) the tumor-associated antigen binding moiety is linked to the N-terminus of the first portion of the dimerization module, and the at least one cytokine molecule or functional fragment or functional variant thereof is linked to the C-terminus of the first portion of the dimerization module, the N- terminus of the second portion of the dimerization module, the C-terminus of the second portion of the dimerization module, or any combination thereof; or (ii) the tumor-associated antigen binding moiety is linked to the C-terminus of the first portion of the dimerization module, and the at least one cytokine molecule or functional fragment or functional variant thereof is linked to the N-terminus of the first portion of the dimerization module, the N- terminus of the second portion of the dimerization module, the C-terminus of the second portion of the dimerization module, or any combination thereof.WSGR Docket No.53676-769.601 301. The multifunctional molecule of claim 299 or 300, wherein the TCRV-binding moiety and the at least one cytokine molecule or functional fragment or functional variant thereof is within a single contiguous polypeptide chain of the first polypeptide or the second polypeptide.

302. The multifunctional molecule of any one of claims 294-301, wherein the tumor-associated antigen binding moiety, the TCRV-binding moiety, or a combination thereof comprises an antibody or antigen binding fragment thereof, wherein the antigen binding fragment comprises any one selected from the group consisting of a Fab, a F(ab')2, an Fv, a single chain Fv (scFv), a single domain antibody, a diabody, a camelid antibody, and a combination thereof.

303. The multifunctional molecule of any one of claims 294-302, wherein the TCRV-binding moiety comprises a heavy chain variable domain (VH) and a light chain variable domain (VL), or a single domain antibody.

304. The multifunctional molecule of any one of claims 299-302, wherein the TCRV-binding moiety comprises a first portion of the TCRV-binding moiety, and wherein the multifunctional molecule further comprises a third polypeptide comprising a second portion of the TCRV-binding moiety, wherein the third polypeptide is non-contiguous with the first polypeptide and the second polypeptide.

305. The multifunctional molecule of claim 304, wherein the first portion of the TCRV-binding moiety comprises a VH of the TCRV-binding moiety and the second portion of the TCRV-binding moiety comprises a VL of the TCRV-binding moiety, or the first portion of the TCRV-binding moiety comprises a VL of the TCRV-binding moiety and the second portion of the TCRV-binding moiety comprises a VH of the TCRV-binding moiety.

306. The multifunctional molecule of any one of claims 294-305, wherein the tumor-associated antigen binding moiety comprises a VH and a VL, or a single domain antibody.

307. The multifunctional molecule of any one of claims 299-305, wherein the tumor-associated antigen binding moiety comprises a first portion of the tumor-associated antigen binding moiety, and wherein the multifunctional molecule further comprises a fourth polypeptide comprising a second portion of the tumor-associated antigen binding moiety, wherein the fourth polypeptide is non- contiguous with the first polypeptide, the second polypeptide, and the third polypeptide.

308. The multifunctional molecule of claim 307, wherein the first portion of the tumor-associated antigen binding moiety comprises a VH of the tumor-associated antigen binding moiety and the second portion of the tumor-associated antigen binding moiety comprises a VL of the tumor-associated antigen binding moiety, or the first portion of the tumor-associated antigen binding moiety comprises a VL of the tumor-associated antigen binding moiety and the second portion of the tumor-associated antigen binding moiety comprises a VH of the tumor-associated antigen binding moiety.

309. The multifunctional molecule of any one of claims 294-308, wherein the first portion of the dimerization module and the second portion of the dimerization module are dimerized.

310. The multifunctional molecule of any one of claims 294-309, wherein:WSGR Docket No.53676-769.601 (i) the tumor-associated antigen binding moiety further comprises a heavy chain constant domain 1 (CH1) linked to the VH of the tumor-associated antigen binding moiety; (ii) the TCRV-binding moiety further comprises a heavy chain constant domain 1 (CH1) linked to the VH of the TCRV-binding moiety; or (iii) a combination thereof.

311. The multifunctional molecule of any one of claims 294-310, wherein: (i) the tumor-associated antigen binding moiety further comprises a light chain constant domain (CL) linked to the VL of the tumor-associated antigen binding moiety; (ii) the TCRV-binding moiety further comprises a light chain constant domain (CL) linked to the VL of the TCRV-binding moiety; or (iii) a combination thereof.

312. The multifunctional molecule of claim 311, wherein: (i) the CL linked to the VL of the tumor-associated antigen binding moiety comprises a kappa chain constant domain or a lambda chain constant domain; (ii) the CL linked to the VL of the TCRV-binding moiety comprises a kappa chain constant domain or a lambda chain constant domain; or (iii) a combination thereof.

313. The multifunctional molecule of claim 312, wherein the kappa chain constant domain or the lambda chain constant domain comprises any one of the light chain constant region sequences listed in Table 3.

314. The multifunctional molecule of any one of claims 294-313, further comprising: (i) a linker between the first portion of the dimerization module and the tumor-associated antigen binding moiety or the first portion of the tumor-associated antigen; (ii) a linker between the at least one cytokine molecule or functional fragment or functional variant thereof and the first portion of the dimerization module, a linker between the at least one cytokine molecule or functional fragment or functional variant thereof and the second portion of the dimerization module, or a combination thereof; (iii) a linker between the at least one cytokine molecule or functional fragment or functional variant thereof and the TCRV-binding moiety or the first portion of the TCRV-binding moiety; (iv) a linker between the VH and the VL of the tumor-associated antigen binding moiety; (v) a linker between the VH and the VL of the TCRV-binding moiety; (vi) a linker between the CH1 and the VH of the tumor-associated antigen binding moiety; (vii) a linker between the CH1 and the VH of the TCRV-binding moiety; (viii) a linker between the CL and the VL of the tumor-associated antigen binding moiety; (ix) a linker between the CL and the VL of the TCRV-binding moiety; or (x) any combination thereof.WSGR Docket No.53676-769.601 315. The multifunctional molecule of claim 314, 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.

316. The multifunctional molecule of claim 315, wherein the linker is the peptide linker, and wherein the linker comprises the sequence of SEQ ID NO: 3308 or SEQ ID NO: 3643.

317. The multifunctional molecule of any one of claims 294-316, wherein the multifunctional molecule is an isolated multifunctional molecule .

318. The multifunctional molecule of any one of claims 294-317, wherein the tumor-associated antigen binding moiety, the TCRV-binding moiety, or a combination thereof comprises the Fab or the scFv.

319. The multifunctional molecule of any one of claims 294-318, wherein the at least one cytokine molecule or functional fragment or functional variant thereof is selected from the group consisting of interleukin-2 (IL-2) or functional fragment or functional variant thereof, interleukin-7 (IL-7) or functional fragment or functional variant thereof, interleukin-12 (IL-12) or functional fragment or functional variant thereof, interleukin-15 (IL-15) or functional fragment or functional variant thereof, interleukin-18 (IL-18) or functional fragment or functional variant thereof, interleukin-21 (IL-21) or functional fragment or functional variant thereof, or interferon gamma or functional fragment or functional variant thereof, or a combination thereof.

320. The multifunctional molecule of any one of claims 294-318, wherein the at least one cytokine molecule or functional fragment or functional variant thereof comprises interleukin-2 (IL-2) or functional fragment or functional variant thereof.

321. The multifunctional molecule of claim 320, wherein the at least one cytokine molecule or functional fragment or functional variant thereof is an IL-2 variant comprising a substitution mutation.

322. The multifunctional molecule of claim 321, wherein the at least one cytokine molecule or functional fragment or functional variant thereof is an IL-2 variant comprising C125A mutation.

323. The multifunctional molecule of any one of claims 320-322, wherein the at least one cytokine molecule or functional fragment or functional variant thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 2270.

324. The multifunctional molecule of any one of claims 320-322, wherein the at least one cytokine molecule or functional fragment or functional variant thereof comprises the sequence of SEQ ID NO: 2270.

325. The multifunctional molecule of any one of claims 294-324, wherein the first portion of the dimerization module comprises a first immunoglobulin constant regions (Fc regions) and the second portion of the dimerization module comprises a second Fc region.

326. The multifunctional molecule of claim 325, wherein the first Fc region, the second Fc region, or a combination thereof is selected from the group consisting of an IgG1 Fc region or a fragment thereof, an IgG2 Fc region or a fragment thereof, an IgG3 Fc region or a fragment thereof, an IgGA1 Fc region or a fragment thereof, an IgGA2 Fc region or a fragment thereof, an IgG4 Fc region or aWSGR Docket No.53676-769.601 fragment thereof, an IgJ Fc region or a fragment thereof, an IgM Fc region or a fragment thereof, an IgD Fc region or a fragment thereof, and an IgE Fc region or a fragment thereof.

327. The multifunctional molecule of claim 326, wherein the first Fc region, the second Fc region, or a combination thereof is selected from the group consisting of a human IgG1 Fc region or a fragment thereof, a human IgG2 Fc region or a fragment thereof, and a human IgG4 Fc region or a fragment thereof.

328. The multifunctional molecule of any one of claims 325-327, 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.

329. The multifunctional molecule of claim 328, wherein the first Fc region, the second Fc region, or a combination thereof comprises an amino acid substitution listed in Table 14.

330. The multifunctional molecule of claim 328 or 329, wherein the first Fc region, the second Fc region, or a combination thereof comprises an Asn297Ala (N297A) mutation or a Leu234Ala / Leu235Ala (LALA) mutation.

331. The multifunctional molecule of any one of claims 328-330, wherein the first Fc region, the second Fc region, or a combination thereof comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 3452, SEQ ID NO: 3454, SEQ ID NO: 3645, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO:3648, SEQ ID NO: 3649, SEQ ID NO: 3792, or SEQ ID NO: 3794.

332. The multifunctional molecule of any one of claims 328-331, wherein the first Fc region, the second Fc region, or a combination thereof comprises the sequence of SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 206, SEQ ID NO: 207, SEQ ID NO: 3452, SEQ ID NO: 3454, SEQ ID NO: 3645, SEQ ID NO: 3646, SEQ ID NO: 3647, SEQ ID NO:3648, SEQ ID NO: 3649, SEQ ID NO: 3792, or SEQ ID NO: 3794.

333. The multifunctional molecule of any one of claims 328-332, wherein the first Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3792, and the second Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, or the first Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, and the second Fc region comprises a sequence having at least 75% sequence identity to the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794.

334. The multifunctional molecule of any one of claims 328-333, wherein the first Fc region comprises a sequence having the sequence of SEQ ID NO: 3649 or SEQ ID NO: 3792, and the second Fc region comprises the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, or the first Fc region comprisesWSGR Docket No.53676-769.601 the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794, and the second Fc region comprises the sequence of SEQ ID NO: 3648 or SEQ ID NO: 3794.

335. The multifunctional molecule of any one of claims 294-334, wherein the TCRV-binding moiety binds to one or more of a TCRβV subfamily.

336. The multifunctional molecule of any one of claims 294-335, wherein the TCRV-binding moiety binds to one or more of a TCRβV subfamily selected from the group consisting of TCRβ V2 subfamily, TCRβ V3 subfamily, TCRβ V4 subfamily, TCRβ V5 subfamily, TCRβ V6 subfamily, TCRβ V9 subfamily, TCRβ V10 subfamily, TCRβ V11 subfamily, TCRβ V12 subfamily, TCRβ V13 subfamily, TCRβ V16 subfamily, TCRβ V19, TCRβ V20 subfamily, TCRβ V21 subfamily, TCRβ V23 subfamily, TCRβ V27 subfamily, and TCRβ V28 subfamily.

337. The multifunctional molecule of any one of claims 294-336, wherein the TCRV-binding moiety binds to one or more of a TCRβV subfamily selected from the group consisting of: (i) TCRβ V2 subfamily comprising TCRβ V2*01; (ii) TCRβ V3 subfamily comprising TCRβ V3-1*01; (iii) TCRβ V4 subfamily comprising one or more selected from TCRβ V4-1, TCRβ V4-2, and TCRβ V4-3; (iv) TCRβ V5 subfamily comprising one or more selected from TCRβ V5-6*01, TCRβ V5-4*01, TCRβ V5-1*01, and TCRβ V5-8*01; (v) TCRβ V6 subfamily comprising one or more selected from 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, and TCRβ V6-1*01; (vi) TCRβ V10 subfamily comprising one or more selected from TCRβ V10-1*01, TCRβ V10- 1*02, TCRβ V10-3*01, and TCRβ V10-2*01; (vii) TCRβ V11 subfamily comprising TCRβ V11-2; (viii) TCRβ V12 subfamily comprising one or more selected from TCRβ V12-4*01, TCRβ V12- 3*01, and TCRβ V12-5*01; (ix) TCRβ V13 subfamily comprising TCRβ V13*01; (x) TCRβ V16 subfamily comprising TCRβ V16*01; (xi) TCRβ V19 subfamily comprising one or more selected from TCRβ V19*01 and TCRβ V19*02; or (xii) TCRβ V20 subfamily comprising TCRβ V20-1*01, or TCRβ V20-1*02.

338. The multifunctional molecule of any one of claims 294-337, wherein the TCRV-binding moiety binds to TCRβ V6 subfamily or TCRβ V20 subfamily.

339. The multifunctional molecule of any one of claims 335-338, wherein the TCRV-binding moiety is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRβV region.

340. The multifunctional molecule of claim 339, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variableWSGR Docket No.53676-769.601 fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

341. The multifunctional molecule of any one of claims 335-340, wherein the TCRV-binding moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

342. The multifunctional molecule of any one of claims 335-341, wherein the TCRV-binding moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

343. The multifunctional molecule of any one of claims 335-342, wherein the TCRV-binding moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

344. The multifunctional molecule of any one of claims 335-343, wherein the TCRV-binding moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 1, 2, 10B, 10C, 11-13, 15-21, 23, or 25; or (iii) a combination thereof.

345. The multifunctional molecule of any one of claims 294-334, wherein the TCRV-binding moiety binds to one or more of a TCRαV subfamily.

346. The multifunctional molecule of any one of claims 294-334 and 345, wherein the TCRV-binding moiety binds to one or more of a one or more of a TCRαV subfamily selected from the group consisting of: a TCRαV1 subfamily, a TCRα V2 subfamily, a TCRα V3 subfamily, a TCRα V4WSGR Docket No.53676-769.601 subfamily, a TCRα V5 subfamily, a TCRα V6 subfamily, a TCRα V7 subfamily, a TCRα V8 subfamily, a TCRα V9 subfamily, a TCRα V10 subfamily, a TCRα V12 subfamily, a TCRα V13 subfamily, a TCRα V14 subfamily, a TCRα V16 subfamily, a TCRα V17 subfamily, a TCRα V18 subfamily, a TCRα V19 subfamily, a TCRα V20 subfamily, a TCRα V21 subfamily, a TCRα V22 subfamily, a TCRα V23 subfamily, a TCRα V24 subfamily, TCRα V25 subfamily, a TCRα V26 subfamily, a TCRα V27 subfamily, a TCRα V29 subfamily, a TCRα V30 subfamily, a TCRα V34 subfamily, a TCRα V35 subfamily, a TCRα V36 subfamily, a TCRα V38 subfamily, a TCRα V39 subfamily, a TCRα V40 subfamily, and a TCRα V41 subfamily.

347. The multifunctional molecule of any one of claims 294-334, 345, and 346, wherein the TCRV- binding moiety binds to one or more of a TCRαV subfamily selected from the group consisting of: (i) TCRαV1 subfamily comprising one or more selected from TCRα V1-1*01, TCRα V1-1*02, TCRα V1-2*01, and TCRα V1-2*02; (ii) TCRα V2 subfamily comprising one or more selected from TCRα V2*01 and TCRα V2*02; (iii) TCRα V3 subfamily comprising TCRα V3*01; (iv) TCRα V4 subfamily comprising TCRα V4*01; (v) TCRα V5 subfamily comprising TCRα V5*01 (vi) TCRα V6 subfamily comprising one or more selected from TCRα V6*01, TCRα V6*02, TCRα V6*03, TCRα V6*04, TCRα V6*05, and TCRα V6*06; (vii) TCRα V7 subfamily comprising TCRα V7*01; (viii) TCRα V8 subfamily comprising one or more selected from TCRα V8-1*01, TCRα V8-1*02, TCRα V8-2*01, TCRα V8-2*02, TCRα V8-3*01, TCRα V8-3*02, TCRα V8-3*03, TCRα V8- 4*01, TCRα V8-4*02, TCRα V8-4*03, TCRα V8-4*04, TCRα V8-4*05, TCRα V8-4*06, TCRα V8-4*07, TCRα V8-6*01, TCRα V8-6*02, and TCRα V8-7*01; (ix) TCRα V9 subfamily comprising one or more selected from TCRα V9-1*01, TCRα V9-2*01, TCRα V9-2*02, TCRα V9-2*03, and TCRα V9-2*04; (x) TCRα V10 subfamily comprising TCRα V10*01; (xi) TCRα V12 subfamily comprising one or more selected from TCRα V12-1*01, TCRα V12-1*02, TCRα V12-2*01, TCRα V12-2*02, TCRα V12-2*03, TCRα V12-3*01, and TCRα V12-3*02; (xii) TCRα V13 subfamily comprising one or more selected from TCRα V13-1*01, TCRα V13-1*02, TCRα V13-1*03, TCRα V13-2*01, and TCRα V13-2*02; (xiii) TCRα V14 subfamily comprising one or more selected from TCRα V14*01, TCRα V14*02, TCRα V14*03, and TCRα V14*04; (xiv) TCRα V16 subfamily comprising TCRα V16*01; (xv) TCRα V17 subfamily comprising TCRα V17*01; (xvi) TCRα V18 subfamily comprising TCRα V18*01; (xvii) TCRα V19 subfamily comprising TCRα V19*01 (xviii) TCRα V20 subfamily comprising one or more selected from TCRα V20*01, TCRα V20*02, TCRα V20*03, and TCRα V20*04;WSGR Docket No.53676-769.601 (xix) TCRα V21 subfamily comprising one or more selected from TCRα V21*01 and TCRα V20*02; (xx) TCRα V22 subfamily comprising TCRα V22*01; (xxi) TCRα V23 subfamily comprising one or more selected from TCRα V23*01, TCRα V23*02, TCRα V23*03, and TCRα V23*04; (xxii) TCRα V24 subfamily comprising one or more selected from TCRα V23*01 and TCRα V23*02; (xxiii) TCRα V25 subfamily comprising TCRα V25*01; (xxiv) TCRα V26 subfamily comprising one or more selected from TCRα V26-1*01, TCRα V26- 1*02, TCRα V26-1*03, TCRα V26-2*01, and TCRα V26-2*02; (xxv) TCRα V27 subfamily comprising one or more selected from TCRα V27*01, TCRα V27*02, and TCRα V27*03; (xxvi) TCRα V29 subfamily comprising one or more selected from TCRα V29*01 and TCRα V27*02; (xxvii) TCRα V30 subfamily comprising one or more selected from TCRα V30*01, TCRα V30*02, TCRα V30*03, and TCRα V30*04; (xxviii) TCRα V34 subfamily comprising TCRα V34*01; (xxix) TCRα V35 subfamily comprising one or more selected from TCRα V35*01 and TCRα V35*02; (xxx) TCRα V36 subfamily comprising one or more selected from TCRα V36*01, TCRα V36*02, TCRα V36*03, and TCRα V36*04; (xxxi) TCRα V38 subfamily comprising one or more selected from TCRα V38-1*01, TCRα V38- 1*02, TCRα V38-1*03, TCRα V38-1*04, and TCRα V38-2*01; (xxxii) TCRα V39 subfamily comprising TCRα V39*01; (xxxiii) TCRα V40 subfamily comprising TCRα V40*01; and (xxxiv) TCRα V41 subfamily comprising TCRα V41*01.

348. The multifunctional molecule of any one of claims 294-334 and 345-347, wherein the TCRV- binding moiety is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to a TCRαV region.

349. The multifunctional molecule of claim 348, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

350. The multifunctional molecule of any one of claims 345-349, wherein the TCRV-binding moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Table 22; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Table 22; orWSGR Docket No.53676-769.601 (iii) a combination thereof.

351. The multifunctional molecule of any one of claims 345-350, wherein the TCRV-binding moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

352. The multifunctional molecule of any one of claims 345-351, wherein the TCRV-binding moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

353. The multifunctional molecule of any one of claims 345-352, wherein the TCRV-binding moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Table 22; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Table 22; or (iii) a combination thereof.

354. The multifunctional molecule of any one of claims 294-353, wherein the multifunctional molecule consists of a first polypeptide and a second polypeptide, wherein the first polypeptide chain comprises the tumor-associated antigen binding moiety linked to an Fc region, wherein the tumor- associated antigen binding moiety is an scFv that binds to a tumor-associated antigen.

355. The multifunctional molecule of any one of claims 294-353, wherein the multifunctional molecule consists of a first polypeptide, a second polypeptide, and a third polypeptide, wherein the first polypeptide chain comprises a first portion of the tumor-associated antigen binding moiety linked to an Fc region, wherein the tumor-associated antigen binding moiety is an Fab that binds to a tumor- associated antigen; and the third polypeptide comprises a second portion of the tumor-associated antigen binding moiety.

356. The multifunctional molecule of any one of claims 294-355, wherein the tumor-associated antigen binding moiety is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to DLL3.

357. The multifunctional molecule of claim 356, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variableWSGR Docket No.53676-769.601 fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

358. The multifunctional molecule of claim 356 or 357, wherein the tumor-targeting moiety comprises: (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 34 or 35; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 34 or 35; or (iii) a combination thereof.

359. The multifunctional molecule of any one of claims claim 356-358, wherein the tumor-targeting moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 34 or 35; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 34 or 35; or (iii) a combination there of.

360. The multifunctional molecule of any one of claims claim 356-359, wherein the tumor-targeting moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 34 or 35; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 34 or 35; or (iii) a combination thereof.

361. The multifunctional molecule of any one of claims claim 356-360, wherein the tumor-targeting moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 34 or 35; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 34 or 35; or (iii) a combination thereof.

362. The multifunctional molecule of any one of claims 294-355, wherein the tumor-associated antigen binding moiety is a full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof, a bivalent antibody, a bispecific antibody, or a biparatopic antibody that binds to CLDN18.

2.

363. The multifunctional molecule of claim 362, wherein the full antibody, an antigen binding domain thereof, or a functional fragment or variant thereof is a Fab, a Fab', a F(ab’)2, a F(ab)2, a variable fragment (Fv), a single domain antibody, a diabody, a nanobody, a camelid single domain antibody, or a single chain variable fragment (scFv).

364. The multifunctional molecule of claim 362 or 363, wherein the tumor-targeting moiety comprises:WSGR Docket No.53676-769.601 (i) a VH comprising a combination of a HC CDR1, a HC CDR2 and a HC CDR3 listed in Tables 38 or 39; (ii) a VL comprising a combination of a LC CDR1, a LC CDR2, and a LC CDR3 listed in Tables 38 or 39; or (iii) a combination thereof.

365. The multifunctional molecule of any one of claims claim 362-364, wherein the tumor-targeting moiety comprises: (i) a VH comprising a HC CDR1, a HC CDR2 and a HC CDR3 of any one of the heavy chain variable domain amino acid sequences listed in Tables 38 or 39; (ii) a VL comprising a LC CDR1, a LC CDR2, and a LC CDR3 of any one of the light chain variable domain amino acid sequences listed in Tables 38 or 39; or (iii) a combination there of.

366. The multifunctional molecule of any one of claims claim 362-365, wherein the tumor-targeting moiety comprises: (i) a VH comprising a sequence having at least 70% sequence identity to any one of the heavy chain variable domain amino acid sequences listed in Tables 38 or 39; (ii) a VL comprising a sequence having at least 70% sequence identity to any one of the light chain variable domain amino acid sequences listed in Tables 38 or 39; or (iii) a combination thereof.

367. The multifunctional molecule of any one of claims claim 362-366, wherein the tumor-targeting moiety comprises: (i) a VH comprising any one of the heavy chain variable domain amino acid sequences listed in Tables 38 or 39; (ii) a VL comprising any one of the light chain variable domain amino acid sequences listed in Tables 38 or 39; or (iii) a combination thereof.

368. The multifunctional molecule of any one of claims 294-367, wherein the multifunctional molecule is a polypeptide molecule.

369. The multifunctional molecule of any one of claims 294-368, wherein the multifunctional molecule is a multispecific molecule.

370. A polynucleotide comprising a sequence encoding the multifunctional molecule of any one of claims 294-369.

371. The polynucleotide of claim 370, wherein the polynucleotide is an isolated nucleic acid molecule.

372. A vector comprising one or more of the polynucleotides of claim 370 or 371.

373. A cell comprising the polynucleotide of claim 370 or 371, or the vector of claim 372.

374. A pharmaceutical composition comprising the multifunctional molecule of any one of claims 294- 369, the polynucleotide of claim 370 or 371, the vector of claim 372, or the cell of claim 373, and a pharmaceutically acceptable carrier, excipient, or diluent.WSGR Docket No.53676-769.601 375. A method of treating a condition or disease in a subject in need therefor comprising administering to the subject a therapeutically effective amount of the multifunctional molecule of any one of claims 294-369, the polynucleotide of claim 370 or 371, the vector of claim 372, the cell of claim 373, the pharmaceutical composition of claim 374, or a combination thereof, wherein the administering is effective to treat the condition or disease in the subject.

376. The method of claim 375, wherein the condition or disease is cancer.

377. The method of claim 376, wherein the cancer is a solid tumor, a hematological cancer, a metastatic cancer, a soft tissue tumor, or a combination thereof.

378. The method of claim 377, wherein the cancer is the solid tumor, and wherein the solid tumor is selected from the group consisting of melanoma, pancreatic cancer, breast cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, liver cancer, and a combination thereof.

379. The method of claim 377, wherein the cancer is the hematological cancer, and wherein the hematological cancer is selected from the group consisting of Hodgkin’s lymphoma, Non-Hodgkin’s lymphoma, acute myeloid leukemia (AML), chronic myeloid leukemia, myelodysplastic syndrome, multiple myeloma, T-cell lymphoma, acute lymphocytic leukemia, and a combination thereof.

380. The method of claim 379, wherein the Non-Hodgkin’s lymphoma is selected from the group consisting of B cell lymphoma, diffuse large B cell lymphoma (DLBCL), follicular lymphoma, chronic lymphocytic leukemia (B-CLL), mantle cell lymphoma, marginal zone B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma, hairy cell leukemia, and a combination thereof.

381. The method of claim 379, wherein the T-cell lymphoma is peripheral T-cell lymphoma.

382. The method of any one of claims 376-381, wherein the cancer is characterized by a cancer antigen present on the cancer.

383. The method of claim 382, wherein the cancer antigen is a tumor antigen, a stromal antigen, or a hematological antigen.

384. The method of any one of claims 382-383, wherein the cancer antigen is selected from the group consisting of BCMA, CD19, CD20, CD22, FcRH5, PDL1, CD47, gangloside 2 (GD2), prostate stem cell antigen (PSCA), prostate specific membrane antigen (PMSA), prostate-specific antigen (PSA), carcinoembryonic antigen (CEA), Ron Kinase, c-Met, Immature laminin receptor, TAG-72, BING-4, Calcium-activated chloride channel 2, Cyclin-B1, 9D7, Ep-CAM, EphA3, Her2 / neu, Telomerase, SAP-1, Survivin, NY-ESO-1 / LAGE-1, PRAME, SSX-2, Melan-A / MART-1, Gp100 / pmel17, Tyrosinase, TRP-1 / -2, MC1R, β-catenin, BRCA1 / 2, CDK4, CML66, Fibronectin, p53, Ras, TGF-Β receptor, AFP, ETA, MAGE, MUC-1, CA-125, BAGE, GAGE, NY-ESO-1, β-catenin, CDK4, CDC27, α actinin-4, TRP1 / gp75, TRP2, gp100, Melan-A / MART1, gangliosides, WT1, EphA3, Epidermal growth factor receptor (EGFR), MART-2, MART-1, MUC1, MUC2, MUM1, MUM2, MUM3, NA88-1, NPM, OA1, OGT, RCC, RUI1, RUI2, SAGE, TRG, TRP1, TSTA, Folate receptor alpha, L1-CAM, CAIX, gpA33, GD3, GM2, VEGFR, Intergrins, carbohydrates, IGF1R, EPHA3, TRAILR1, TRAILR2, RANKL, FAP, TGF-beta, hyaluronic acid, collagen, tenascin C, delta-like ligand 3 (DLL3), CLDN18.2, and tenascin W.WSGR Docket No.53676-769.601 385. The method of any one of claims 382-384, wherein the cancer antigen is DLL3.

386. The method of any one of claims 382-384, wherein the cancer antigen is CLDN18.

2.

387. The method of any one of claims 375-386, further comprising administering a second therapeutic agent or therapy to the subject.

388. The method of claim 387, wherein the second therapeutic agent or therapy comprises a chemotherapeutic agent, a biologic agent, a hormonal therapy, radiation, or surgery.

389. The method of claim 387 or 388, wherein the second therapeutic agent or therapy is administered in combination with the multifunctional molecule of any one of claims 294-369, the polynucleotide of claim 370 or 371, the vector of claim 372, the cell of claim 373, or the pharmaceutical composition of claim 374, sequentially, simultaneously, or concurrently.

390. Use of the multifunctional molecule of any one of claims 294-369, the polynucleotide of claim 370 or 371, the vector of claim 372, the cell of claim 373, or the pharmaceutical composition of claim 374 for the method of any one of claims 375-389.

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