Chimeric antigen receptor targeting G-protein coupled receptor and its use

Abstract

The subject matter disclosed in the present invention provides methods and compositions for treating multiple myeloma. The subject matter disclosed in the present invention relates to chimeric antigen receptors (CARs) specifically targeting G-protein coupled receptors (e.g., G-protein coupled receptor C family 5 group D member (GPRC5D)) and immune response cells comprising such CARs. The CARs targeting G-protein coupled receptors (e.g., GPRC5D) disclosed in the present invention have enhanced immune activation properties, including anti-tumor activity.

Description

[0001] This application is a divisional application of the invention patent application with the invention name “Chimeric antigen receptor targeting G-protein coupled receptor and its use” and application number “201580075557.4” submitted to the State Intellectual Property Office of the People’s Republic of China on August 4, 2017.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 088,286, filed December 5, 2014, which is incorporated herein by reference in its entirety and claims the benefit of priority. Technical Field

[0004] The subject matter disclosed in the present invention provides methods and compositions for treating cancer.It relates to chimeric antigen receptors (CARs) specifically targeting G-protein coupled receptors (e.g., G-protein coupled receptor C family 5 group D member (GPRC5D)), immune response cells comprising such CARs, and methods using such cells for treating cancer (e.g., multiple myeloma). Background Art

[0005] Cell-based immunotherapy is a therapy with curative potential for cancer treatment. T cells and other immune cells are modified to target tumor antigens by introducing genetic material encoding artificial or synthetic receptors for antigens, which are called chimeric antigen receptors (CARs), which are specific for the selected antigens. Targeted T cell therapy using CARs has recently been shown to achieve clinical success in the treatment of hematological malignancies.

[0006] Multiple myeloma (MM) is the second most common hematological malignancy. 9 Approximately 25% of patients have high-risk cytogenetics, which portend a median survival of less than 2 years. 10,11 Despite recent advances, the disease remains incurable outside of the immunotherapeutic graft-versus-myeloma (GvM) effect of allogeneic transplantation, regardless of cytogenetics. However, allogeneic transplantation is limited by ineligibility and high rates of transplant-related morbidity and mortality. 12 Similar to the GvM effect, potentially curative T cell effects can be achieved with minimal toxicity through autologous adoptive T cell therapy.

[0007] Myeloma may be an ideal disease to test adoptive T cell therapy. First, as mentioned above, allogeneic transplants have demonstrated that T cells can be curative, even with minimal or no concomitant chemotherapy, such as after non-myeloablative transplantation or post-transplant donor lymphocyte infusion. Second, conditioning chemotherapy may enhance the efficacy of adoptive T cell therapy by depleting regulatory T cells (Tregs). 5,13 Thus, the immediate post-autologous transplant period may be the optimal time to administer T cells, and myeloma is one of the few diseases where autologous stem cell transplantation is the standard of care. Third, the immunomodulatory drug lenalidomide may improve CAR-based therapies, as has been shown in mice. 14 And lenalidomide is commonly used to treat MM. Fourth, compared with solid tumors or extramedullary CLL, 5 Adoptive T cell therapy is most effective in myeloid-primary diseases such as ALL. 7,8 And like ALL, myeloma is a disease of the bone marrow.

[0008] Although there are various reasons to expect that adoptive T-cell therapy may have a role in MM, extending adoptive T-cell therapy to myeloma also presents unique challenges. Unlike other B-cell malignancies, CD19 expression is seen in only 2% of myeloma patients. 15 Furthermore, unlike CD19, common extracellular immunophenotypic markers in myeloma (CD138, CD38, and CD56) are all co-expressed on other essential cell types, and we predict that CARs directed against any of these targets would result in unacceptable off-tumor, on-target toxicity. 7 , which can be lethal even in targets for which antibodies are well tolerated, as is the case with CARs targeting HER2. 16 To address these challenges, we have identified extracellular targets with predicted high MM expression and limited expression in essential normal tissues that may be optimal targets for adoptive T cell therapy for MM. Therefore, novel therapeutic strategies are needed to design CARs targeting antigens that are highly expressed in MM cells and limitedly expressed in normal tissues for the treatment of multiple myeloma, which can induce effective tumor eradication with minimal toxicity and immunogenicity. Summary of the Invention

[0009] The presently disclosed subject matter generally provides chimeric antigen receptors (CARs) that specifically target G-protein coupled receptors, immune response cells comprising such CARs, and uses of these CARs and immune response cells for treating multiple myeloma.

[0010] The subject matter disclosed herein provides CAR. In a non-limiting example, CAR comprises an extracellular antigen binding domain, a transmembrane domain, and an intracellular domain, wherein the extracellular antigen binding domain specifically binds to a G-protein coupled receptor. In certain embodiments, the G-protein coupled receptor is a G-protein coupled receptor C family 5 group D member (GPRC5D). In certain embodiments, the extracellular antigen binding domain is about 1x10 -9 M is about 3x10 -6 The binding affinity (K D ) specifically binds to GPRC5D. In certain embodiments, the extracellular antigen-binding domain is a single-chain variable fragment (scFv). In certain embodiments, the extracellular antigen-binding domain is a murine scFv. In certain embodiments, the extracellular antigen-binding domain is a human scFv. In certain embodiments, the extracellular antigen-binding domain is an optionally cross-linked Fab. In certain embodiments, the extracellular binding domain is a F(ab)2. In certain embodiments, any of the aforementioned molecules can be included in a fusion protein with a heterologous sequence to form the extracellular antigen-binding domain.

[0011] In certain embodiments, the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374, and 386.

[0012] In certain embodiments, the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375, and 387.

[0013] In certain embodiments, the extracellular antigen binding domain comprises (a) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374, and 386; and (b) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: A light chain variable region of an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence of SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387.

[0014] In certain embodiments, the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374 and 386, and conservative modifications thereof.

[0015] In certain embodiments, the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence selected from SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387 and conservative modifications thereof.

[0016] In certain embodiments, the extracellular antigen binding domain comprises (a) a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374, and 386, and conservative modifications thereof, and (b) a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: The amino acid sequences of IDNO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387 and conservatively modified light chain variable regions thereof.

[0017] In certain embodiments, the extracellular antigen binding domain comprises a heavy chain variable region comprising amino acids having a sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374, and 386. In certain embodiments, the extracellular antigen-binding domain comprises a light chain variable region comprising amino acids having a sequence selected from the group consisting of SEQ ID NOs: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375, and 387. In certain embodiments, the extracellular antigen-binding domain comprises a heavy chain variable region comprising amino acids having a sequence as set forth in SEQ ID NO: 53. In certain embodiments, the extracellular antigen-binding domain comprises a heavy chain variable region comprising amino acids having a sequence as set forth in SEQ ID NO: 57. In certain embodiments, the extracellular antigen-binding domain comprises a heavy chain variable region comprising amino acids having a sequence as set forth in SEQ ID NO: 61. In certain embodiments, the extracellular antigen-binding domain comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 65. In certain embodiments, the extracellular antigen-binding domain comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 69. In certain embodiments, the extracellular antigen-binding domain comprises a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 54. In certain embodiments, the extracellular antigen-binding domain comprises a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 58. In certain embodiments, the extracellular antigen-binding domain comprises a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 62. In certain embodiments, the extracellular antigen-binding domain comprises a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 66. In certain embodiments, the extracellular antigen-binding domain comprises a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 70. In certain embodiments, the extracellular antigen-binding domain comprises: (a) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 1, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 2; (b) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 5, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 6;(c) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 9, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 10; (d) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 13, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 14; (e) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 17, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 18; (f) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 21, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 22; (g) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 25, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 26; (h) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 29, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 30; (i) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 31. NO: 33, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 34; (j) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 37, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 38; (k) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 41, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 42; (l) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 45, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 46; (m) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 49, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 50; (n) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 53, and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 54; (o) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: NO: 57, and a light chain variable region comprising the amino acids having the sequence set forth in SEQ ID NO: 58; (p) a heavy chain variable region comprising the amino acids having the sequence set forth in SEQ ID NO: 61, and a light chain variable region comprising the amino acids having the sequence set forth in SEQ ID NO: 62; (q) a heavy chain variable region comprising the amino acids having the sequence set forth in SEQ ID NO: 65, and a light chain variable region comprising the amino acids having the sequence set forth in SEQ ID NO: 66;(r) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 69, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 70; (s) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 73, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 74; (t) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 77, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 78; (u) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 81, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 82; (v) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 85, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 86; (w) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 89, and a light chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: 90; (x) a heavy chain variable region comprising the amino acids of the sequence set forth in SEQ ID NO: (aa) a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 326, and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 327; (ab) a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 338, and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 339; (ac) a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 350, and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 351. NO: 351; (ad) a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 362, and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 363; (ae) a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 374, and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 375;or (af) a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 386, and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 387. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 53; and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 54. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 57; and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 58. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 61; and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 62. In another embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region comprising the amino acids set forth in SEQ ID NO: 65; and a light chain variable region comprising the amino acids set forth in SEQ ID NO: 66. In yet another embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 69; and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 70.

[0018] In certain embodiments, the extracellular antigen-binding domain comprises both the heavy and light chains, optionally with a linker sequence, such as a linker peptide, between the heavy and light chain variable regions. For example, in certain non-limiting embodiments, the extracellular antigen-binding domain comprises: (i) a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 57 and (ii) a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 58, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy and light chain variable regions. In certain embodiments, the extracellular antigen-binding domain comprises: (i) a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 61 and (ii) a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 62, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy and light chain variable regions. In certain embodiments, the extracellular antigen-binding domain comprises: (i) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 53 and (ii) a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 54, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the extracellular antigen-binding domain comprises: (i) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 61 and (ii) a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 62, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the extracellular antigen-binding domain comprises: (i) a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 65 and (ii) a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 66, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the extracellular antigen-binding domain comprises: (i) a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 69 and (ii) a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 70, optionally with (iii) a linker sequence, e.g., a linker peptide, between the heavy chain variable region and the light chain variable region.

[0019] In certain embodiments, the extracellular antigen binding domain comprises (a) a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222, 228, 234, 240, 246, 252, 258, 264, 306, 318, 330, 342, 354, 366, 378, and 390; and (b) a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: The light chain variable region CDR3 of the amino acid sequence of NO:129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 195, 201, 207, 213, 219, 225, 231, 237, 243, 249, 255, 261, 267, 309, 321, 333, 345, 357, 369, 381 and 393.

[0020] In certain embodiments, the heavy chain variable region CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305, 317, 329, 341, 353, 365, 377, and 389, and conservative modifications thereof; and (b) the light chain variable region CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: The amino acid sequences of NOs: 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 194, 200, 206, 212, 218, 224, 230, 236, 242, 248, 254, 260, 266, 308, 320, 332, 344, 356, 368, 380 and 392 and conservative modifications thereof.

[0021] In certain embodiments, the heavy chain variable region CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 304, 316, 328, 340, 352, 364, 376, and 388, and conservative modifications thereof; and (b) the light chain The variable region CDR1 comprises an amino acid sequence selected from the group consisting of 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 193, 199, 205, 211, 217, 223, 229, 235, 241, 247, 253, 259, 265, 307, 319, 331, 343, 355, 367, 379 and 391 and conservative modifications thereof.

[0022] In certain embodiments, the extracellular antigen binding domain comprises: (a) a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 304, 316, 328, 340, 352, 364, 376, and 388; (b) a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 NO:125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305, 317, 329, 341, 353, 365, 377 and 389; (c) comprising a heavy chain variable region CDR2 selected from the group consisting of SEQ ID NO:126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222, 228, 234, 240, 246, 252, 258, 264, 306, 318, 330, 342, 354, 366, 378 and 390; (d) comprising a heavy chain variable region CDR3 selected from 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 193, 199, 205, 211, 217, 223, 229, 235, 241, 247, 253, 259, 265, 307, 319, 331, 343, 355, 367, 379 and 391; (e) comprising a light chain variable region CDR1 selected from the group consisting of SEQ ID NO: 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 194, 200, 206, 212, 218, 224, 230, 236, 242, 248, 254, 260, 266, 308, 320, 332, 344, 356, 368, 380 and 392; and (f) comprising a light chain variable region CDR2 selected from the group consisting of SEQ ID NO: The light chain variable region CDR3 of the amino acid sequence of NO:129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 195, 201, 207, 213, 219, 225, 231, 237, 243, 249, 255, 261, 267, 309, 321, 333, 345, 357, 369, 381 and 393.

[0023] In certain embodiments, the extracellular antigen-binding domain comprises (a) a heavy chain variable region CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 124 or conservative modifications thereof; a heavy chain variable region CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 125 or conservative modifications thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 126 or conservative modifications thereof; (b) a heavy chain variable region CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 130 or conservative modifications thereof; a heavy chain variable region CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 131 or conservative modifications thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 132 or conservative modifications thereof; (c) a heavy chain variable region CDR1 comprising an amino acid sequence as set forth in SEQ ID NO: 136 or conservative modifications thereof; a heavy chain variable region CDR2 comprising an amino acid sequence as set forth in SEQ ID NO: 137 or conservative modifications thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence as set forth in SEQ ID NO: 138 or conservative modifications thereof. NO: 138 or conservatively modified amino acids thereof; (d) a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 142 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 143 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 144 or conservatively modified amino acids thereof; (e) a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 148 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 149 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 150 or conservatively modified amino acids thereof; (f) a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 154 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 155 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 156 or conservatively modified amino acids thereof NO: 156 or conservatively modified amino acids thereof; (g) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 160 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 161 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence of SEQ ID NO: 162 or conservatively modified amino acids thereof; (h) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 166 or conservatively modified amino acids thereof;comprising a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 167 or conservative modifications thereof; and a heavy chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 168 or conservative modifications thereof; (i) comprising a heavy chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 172 or conservative modifications thereof; comprising a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 173 or conservative modifications thereof; and a heavy chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 174 or conservative modifications thereof; (j) comprising a heavy chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 178 or conservative modifications thereof; comprising a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 179 or conservative modifications thereof; and a heavy chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 180 or conservative modifications thereof; (k) comprising a heavy chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 184 or conservative modifications thereof; comprising a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 185 or conservative modifications thereof NO: 185 or its conservative modifications; and a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 186 or its conservative modifications; (l) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 190 or its conservative modifications; a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 191 or its conservative modifications; and a heavy chain variable region CDR3 comprising a sequence of SEQ ID NO: 192 or its conservative modifications; (m) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 196 or its conservative modifications; a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 197 or its conservative modifications; and a heavy chain variable region CDR3 comprising a sequence of SEQ ID NO: 198 or its conservative modifications; (n) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 202 or its conservative modifications; NO: 203 or conservatively modified amino acids thereof; and a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 204 or conservatively modified amino acids thereof; (o) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 208 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 209 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence of SEQ ID NO: 210 or conservatively modified amino acids thereof;(p) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 214 or conservative modifications thereof; a heavy chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 215 or conservative modifications thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 216 or conservative modifications thereof; (q) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 220 or conservative modifications thereof; a heavy chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 221 or conservative modifications thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 222 or conservative modifications thereof; (r) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 226 or conservative modifications thereof; a heavy chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 227 or conservative modifications thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 228 or conservative modifications thereof; (s) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 229 or conservative modifications thereof NO: 232 or conservatively modified amino acids thereof; a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 233 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 234 or conservatively modified amino acids thereof; (t) a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 238 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 239 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 240 or conservatively modified amino acids thereof; (u) a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 244 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 245 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 246 or conservatively modified amino acids thereof; (v) a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 247 or conservatively modified amino acids thereof NO: 250 or conservatively modified amino acids thereof; a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 251 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence of SEQ ID NO: 252 or conservatively modified amino acids thereof; (w) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 256 or conservatively modified amino acids thereof; and a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 257 or conservatively modified amino acids thereof;and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 258 or a conservative modification thereof; (x) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 262 or a conservative modification thereof; a heavy chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 263 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 264 or a conservative modification thereof; (y) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 304 or a conservative modification thereof; a heavy chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 305 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 306 or a conservative modification thereof; (z) a heavy chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 316 or a conservative modification thereof; a heavy chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 317 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 318 or a conservative modification thereof NO: 318 or conservatively modified amino acids thereof; (aa) comprising a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 328 or conservatively modified amino acids thereof; comprising a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 329 or conservatively modified amino acids thereof; and comprising a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 330 or conservatively modified amino acids thereof; (ab) comprising a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 340 or conservatively modified amino acids thereof; comprising a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 341 or conservatively modified amino acids thereof; and comprising a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 342 or conservatively modified amino acids thereof; (ac) comprising a heavy chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 352 or conservatively modified amino acids thereof; comprising a heavy chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 353 or conservatively modified amino acids thereof; and comprising a heavy chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 354 or conservatively modified amino acids thereof NO: 354 or conservatively modified amino acids thereof; (ad) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 364 or conservatively modified amino acids thereof; a heavy chain variable region CDR2 comprising a sequence of SEQ ID NO: 365 or conservatively modified amino acids thereof; and a heavy chain variable region CDR3 comprising a sequence of SEQ ID NO: 366 or conservatively modified amino acids thereof; (ae) a heavy chain variable region CDR1 comprising a sequence of SEQ ID NO: 376 or conservatively modified amino acids thereof;comprising a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 377 or conservative modifications thereof; and a heavy chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 378 or conservative modifications thereof; or (af) comprising a heavy chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 388 or conservative modifications thereof; comprising a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 389 or conservative modifications thereof; and comprising a heavy chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 390 or conservative modifications thereof. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 202; a heavy chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 203; and a heavy chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 204. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 208; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 209; and a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 210. In another non-limiting embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 214; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 215; and a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 216. In yet another non-limiting embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 220; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 221; and a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 222. In another embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 226; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 227; and a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 228.

[0024] In certain embodiments, the extracellular antigen-binding domain comprises: (a) a light chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 127 or conservative modifications thereof; a light chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 129 or conservative modifications thereof; and a light chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 130 or conservative modifications thereof; (b) a light chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 133 or conservative modifications thereof; a light chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 134 or conservative modifications thereof; and a light chain variable region CDR3 comprising SEQ ID NO: 135 or conservative modifications thereof; (c) a light chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 139 or conservative modifications thereof; a light chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 140 or conservative modifications thereof; and a light chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 141 or conservative modifications thereof. NO: 141 or a conservatively modified amino acid sequence thereof; (d) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 145 or a conservatively modified amino acid sequence thereof; a light chain variable region CDR2 comprising SEQ ID NO: 146 or a conservatively modified amino acid sequence thereof; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 147 or a conservatively modified amino acid sequence thereof; (e) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 151 or a conservatively modified amino acid sequence thereof; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 152 or a conservatively modified amino acid sequence thereof; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 153 or a conservatively modified amino acid sequence thereof; (f) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 157 or a conservatively modified amino acid sequence thereof; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 158 or a conservatively modified amino acid sequence thereof; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 159 or a conservatively modified amino acid sequence thereof. NO: 159 or a conservative modification thereof; (g) a light chain variable region CDR1 comprising the sequence of SEQ ID NO: 163 or a conservative modification thereof; a light chain variable region CDR2 comprising the sequence of SEQ ID NO: 164 or a conservative modification thereof; and a light chain variable region CDR3 comprising the sequence of SEQ ID NO: 165 or a conservative modification thereof; (h) a light chain variable region CDR1 comprising the sequence of SEQ ID NO: 169 or a conservative modification thereof; and a light chain variable region CDR2 comprising the sequence of SEQ ID NO: 170 or a conservative modification thereof;and a light chain variable region CDR3 comprising SEQ ID NO: 171 or a conservative modification thereof; (i) a light chain variable region CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 175 or a conservative modification thereof; a light chain variable region CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 176 or a conservative modification thereof; and a light chain variable region CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 177 or a conservative modification thereof; (j) a light chain variable region CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 181 or a conservative modification thereof; a light chain variable region CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 182 or a conservative modification thereof; and a light chain variable region CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 183 or a conservative modification thereof; (k) a light chain variable region CDR1 comprising an amino acid sequence set forth in SEQ ID NO: 187 or a conservative modification thereof; a light chain variable region CDR2 comprising an amino acid sequence set forth in SEQ ID NO: 188 or a conservative modification thereof; and a light chain variable region CDR3 comprising an amino acid sequence set forth in SEQ ID NO: 189 or a conservative modification thereof NO: 189 or conservatively modified amino acids thereof; (l) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 193 or conservatively modified amino acids thereof; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 194 or conservatively modified amino acids thereof; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 195 or conservatively modified amino acids thereof; (m) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 199 or conservatively modified amino acids thereof; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 200 or conservatively modified amino acids thereof; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 201 or conservatively modified amino acids thereof; (n) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 205 or conservatively modified amino acids thereof; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 206 or conservatively modified amino acids thereof; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 207 or conservatively modified amino acids thereof NO: 207 or conservatively modified amino acids thereof; (o) a light chain variable region CDR1 comprising a sequence of SEQ ID NO: 211 or conservatively modified amino acids thereof; a light chain variable region CDR2 comprising a sequence of SEQ ID NO: 212 or conservatively modified amino acids thereof; and a light chain variable region CDR3 comprising a sequence of SEQ ID NO: 213 or conservatively modified amino acids thereof; (p) a light chain variable region CDR1 comprising a sequence of SEQ ID NO: 217 or conservatively modified amino acids thereof;comprising a light chain variable region CDR2 having an amino acid sequence of SEQ ID NO: 218 or a conservative modification thereof; and a light chain variable region CDR3 having an amino acid sequence of SEQ ID NO: 219 or a conservative modification thereof; (q) comprising a light chain variable region CDR1 having an amino acid sequence of SEQ ID NO: 223 or a conservative modification thereof; comprising a light chain variable region CDR2 having an amino acid sequence of SEQ ID NO: 224 or a conservative modification thereof; and a light chain variable region CDR3 having an amino acid sequence of SEQ ID NO: 225 or a conservative modification thereof; (r) comprising a light chain variable region CDR1 having an amino acid sequence of SEQ ID NO: 229 or a conservative modification thereof; comprising a light chain variable region CDR2 having an amino acid sequence of SEQ ID NO: 230 or a conservative modification thereof; and a light chain variable region CDR3 having an amino acid sequence of SEQ ID NO: 231 or a conservative modification thereof; (s) comprising a light chain variable region CDR1 having an amino acid sequence of SEQ ID NO: 235 or a conservative modification thereof; comprising a light chain variable region CDR2 having an amino acid sequence of SEQ ID NO: 236 or a conservative modification thereof NO: 236 or its conservative modifications; and a light chain variable region CDR2 comprising a sequence of SEQ ID NO: 237 or its conservative modifications; (t) a light chain variable region CDR1 comprising a sequence of SEQ ID NO: 241 or its conservative modifications; a light chain variable region CDR2 comprising a sequence of SEQ ID NO: 242 or its conservative modifications; and a light chain variable region CDR3 comprising a sequence of SEQ ID NO: 243 or its conservative modifications; (u) a light chain variable region CDR1 comprising a sequence of SEQ ID NO: 247 or its conservative modifications; a light chain variable region CDR2 comprising a sequence of SEQ ID NO: 248 or its conservative modifications; and a light chain variable region CDR3 comprising a sequence of SEQ ID NO: 249 or its conservative modifications; (v) a light chain variable region CDR1 comprising a sequence of SEQ ID NO: 253 or its conservative modifications; NO: 254 or a conservative modification thereof; and a light chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 255 or a conservative modification thereof; (w) a light chain variable region CDR1 comprising an amino acid sequence of SEQ ID NO: 259 or a conservative modification thereof; a light chain variable region CDR2 comprising an amino acid sequence of SEQ ID NO: 260 or a conservative modification thereof; and a light chain variable region CDR3 comprising an amino acid sequence of SEQ ID NO: 261 or a conservative modification thereof;(x) comprising a light chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 265 or conservative modifications thereof; a light chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 266 or conservative modifications thereof; and a light chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 267 or conservative modifications thereof; (y) comprising a light chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 307 or conservative modifications thereof; a light chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 308 or conservative modifications thereof; and a light chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 309 or conservative modifications thereof; (z) comprising a light chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 319 or conservative modifications thereof; a light chain variable region CDR2 having an amino acid sequence as set forth in SEQ ID NO: 320 or conservative modifications thereof; and a light chain variable region CDR3 having an amino acid sequence as set forth in SEQ ID NO: 321 or conservative modifications thereof; (aa) comprising a light chain variable region CDR1 having an amino acid sequence as set forth in SEQ ID NO: 319 or conservative modifications thereof; NO: 331 or its conservatively modified amino acids; a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 332 or its conservatively modified amino acids; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 333 or its conservatively modified amino acids; (ab) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 343 or its conservatively modified amino acids; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 344 or its conservatively modified amino acids; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 345 or its conservatively modified amino acids; (ac) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 355 or its conservatively modified amino acids; a light chain variable region CDR2 comprising a sequence as shown in SEQ ID NO: 356 or its conservatively modified amino acids; and a light chain variable region CDR3 comprising a sequence as shown in SEQ ID NO: 357 or its conservatively modified amino acids; (ad) a light chain variable region CDR1 comprising a sequence as shown in SEQ ID NO: 358 or its conservatively modified amino acids. NO: 367 or conservatively modified amino acids thereof; a light chain variable region CDR1 comprising a light chain variable region CDR2 comprising a light chain variable region CDR3 comprising a light chain variable region CDR4 comprising a light chain variable region CDR5 comprising a light chain variable region CDR6 comprising a light chain variable region CDR7 comprising a light chain variable region CDR8 comprising a light chain variable region CDR9 comprising a light chain variable region CDR10 comprising a light chain variable region CDR11 comprising a light chain variable region CDR12 comprising a light chain variable region CDR13 comprising a light chain variable region CDR14 comprising a light chain variable region CDR15 comprising a light chain variable region CDR16 comprising a light chain variable region CDR17 comprising a light chain variable region CDR18 comprising a light chain variable region CDR19 comprising a light chain variable region CDR210 comprising a light chain variable region CDR11 comprising a light chain variable region CDR12 comprising a light chain variable region CDR13 comprising a light chain variable region CDR14 comprising a light chain variable region CDR15 comprising a light chain variable region CDR16 comprising a light chain variable region CDR17 comprising a light chain variable region CDR18 comprising a light chain variable region CDR19 comprising a light chain variable region CDR1 ...and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 381 or conservative modifications thereof; or (af) a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 391 or conservative modifications thereof; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 392 or conservative modifications thereof; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 393 or conservative modifications thereof. In certain embodiments, the extracellular antigen-binding domain comprises: a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 205; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 206; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 207. In certain embodiments, the extracellular antigen-binding domain comprises: a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 211; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 212; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 213. In certain embodiments, the extracellular antigen-binding domain comprises: a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 217; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 218; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 219. In another non-limiting embodiment, the extracellular antigen-binding domain comprises: a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 223; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 224; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 225. In another non-limiting embodiment, the extracellular antigen-binding domain comprises: a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 229; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 230; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 231.

[0025] In certain embodiments, the extracellular antigen-binding domain comprises: (a) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 124; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 125; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 126; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 127; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 128; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 129; (b) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 130; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 131; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 132; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 133; and a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 139; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 140; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 141; (d) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 142; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 143; and a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 144. NO: 144; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 145; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 146; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 147; (e) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 148; and a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 149;a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 150; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 151; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 152; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 153; (f) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 154; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 155; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 156; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 157; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 158; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 159; (g) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 154; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 155; NO: 160; a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 161; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 162; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 163; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 164; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 165; (h) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 166; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 167; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 168; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 169; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 170; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 171. (i) a heavy chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 172; a heavy chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 173; a heavy chain variable region CDR3 comprising the amino acids set forth in SEQ ID NO: 174; a light chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 175; and a light chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 176;and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 177; (j) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 178; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 179; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 180; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 181; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 182; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 183; (k) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 184; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 185; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 186; and a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 187; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 188; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 189; (l) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 190; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 191; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 192; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 193; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 194; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 195; (m) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 196; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 197; NO: 197; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 198; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 199; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 200; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 201; (n) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 202; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 203;a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 204; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 205; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 206; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 207; (o) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 208; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 209; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 210; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 211; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 212; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 213; (p) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 214; NO: 214; a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 215; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 216; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 217; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 218; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 219; (q) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 220; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 221; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 222; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 223; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 224; and (r) a heavy chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 226; a heavy chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 227; a heavy chain variable region CDR3 comprising the amino acids set forth in SEQ ID NO: 228; a light chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 229; and a light chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 230;and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 231; (s) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 232; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 233; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 234; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 235; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 236; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 237; (t) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 238; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 239; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 240; and NO: 241; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 242; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 243; (u) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 244; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 245; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 246; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 247; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 248; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 249; (v) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 250; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 251; NO: 251; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 252; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 253; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 254; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 255; (w) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 256; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 257;a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 258; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 259; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 260; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 261; (x) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 262; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 263; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 264; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 265; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 266; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 267; (y) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 268; NO: 304; a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 305; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 306; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 307; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 308; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 309; (z) a heavy chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 316; a heavy chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 317; a heavy chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 318; a light chain variable region CDR1 comprising the amino acids of the sequence set forth in SEQ ID NO: 319; a light chain variable region CDR2 comprising the amino acids of the sequence set forth in SEQ ID NO: 320; and a light chain variable region CDR3 comprising the amino acids of the sequence set forth in SEQ ID NO: 321. (aa) a heavy chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 328; a heavy chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 329; a heavy chain variable region CDR3 comprising the amino acids set forth in SEQ ID NO: 330; a light chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 331; and a light chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 332.and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 333; (ab) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 340; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 341; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 342; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 343; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 344; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 345; (ac) a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 352; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 353; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 354; and NO: 355; a light chain variable region CDR1 comprising amino acids of the sequence set forth in SEQ ID NO: 356; and a light chain variable region CDR3 comprising amino acids of the sequence set forth in SEQ ID NO: 357; (ad) a heavy chain variable region CDR1 comprising amino acids of the sequence set forth in SEQ ID NO: 364; a heavy chain variable region CDR2 comprising amino acids of the sequence set forth in SEQ ID NO: 365; a heavy chain variable region CDR3 comprising amino acids of the sequence set forth in SEQ ID NO: 366; a light chain variable region CDR1 comprising amino acids of the sequence set forth in SEQ ID NO: 367; a light chain variable region CDR2 comprising amino acids of the sequence set forth in SEQ ID NO: 368; and a light chain variable region CDR3 comprising amino acids of the sequence set forth in SEQ ID NO: 369; (ae) a heavy chain variable region CDR1 comprising amino acids of the sequence set forth in SEQ ID NO: 376; a heavy chain variable region CDR2 comprising amino acids of the sequence set forth in SEQ ID NO: 377; NO: 377; a heavy chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 378; a light chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 379; a light chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 380; and a light chain variable region CDR3 comprising the amino acids set forth in SEQ ID NO: 381; or (af) a heavy chain variable region CDR1 comprising the amino acids set forth in SEQ ID NO: 388; a heavy chain variable region CDR2 comprising the amino acids set forth in SEQ ID NO: 389;comprising a heavy chain variable region CDR3 having the amino acid sequence of SEQ ID NO: 390; a light chain variable region CDR1 having the amino acid sequence of SEQ ID NO: 391; a light chain variable region CDR2 having the amino acid sequence of SEQ ID NO: 392; and a light chain variable region CDR3 having the amino acid sequence of SEQ ID NO: 393. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 having the amino acid sequence of SEQ ID NO: 202; a heavy chain variable region CDR2 having the amino acid sequence of SEQ ID NO: 203; a heavy chain variable region CDR3 having the amino acid sequence of SEQ ID NO: 204; a light chain variable region CDR1 having the amino acid sequence of SEQ ID NO: 205; a light chain variable region CDR2 having the amino acid sequence of SEQ ID NO: 206; and a light chain variable region CDR3 having the amino acid sequence of SEQ ID NO: 207. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence shown in SEQ ID NO: 208; a heavy chain variable region CDR2 comprising amino acids having the sequence shown in SEQ ID NO: 209; a heavy chain variable region CDR3 comprising amino acids having the sequence shown in SEQ ID NO: 210; a light chain variable region CDR1 comprising amino acids having the sequence shown in SEQ ID NO: 211; a light chain variable region CDR2 comprising amino acids having the sequence shown in SEQ ID NO: 212; and a light chain variable region CDR3 comprising amino acids having the sequence shown in SEQ ID NO: 213. In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 214; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 215; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 216; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 217; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 218; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 219. In another non-limiting embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 220; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 221; and a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 222.A light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 223; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 224; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 225. In yet another non-limiting embodiment, the extracellular antigen-binding domain comprises: a heavy chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 226; a heavy chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 227; a heavy chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 228; a light chain variable region CDR1 comprising amino acids having the sequence set forth in SEQ ID NO: 229; a light chain variable region CDR2 comprising amino acids having the sequence set forth in SEQ ID NO: 230; and a light chain variable region CDR3 comprising amino acids having the sequence set forth in SEQ ID NO: 231. In certain embodiments, the human scFv comprises a heavy chain variable region, a light chain variable region, a linker peptide between the heavy chain variable region and the light chain variable region, and a His-tag and an HA-tag. In certain embodiments, the amino acid sequence of the His-tag and the HA-tag comprises the amino acid sequence of SEQ ID NO: 275, which is provided below:

[0026]

[0027] The nucleotide sequence encoding SEQ ID NO:275 is SEQ ID NO:276, which is provided below:

[0028]

[0029] In certain embodiments, GPRC5D comprises the amino acid sequence set forth in SEQ ID NO: 97. In certain embodiments, the extracellular antigen-binding domain binds to one, two, three, or four epitope regions selected from the group consisting of an epitope region within the N-terminal region encompassing amino acids 1-27 of SEQ ID NO: 97, an epitope region within the ECL1 region encompassing amino acids 85-93 of SEQ ID NO: 97, an epitope region within the ECL2 region encompassing amino acids 145-167 of SEQ ID NO: 97, and an epitope region within the ECL3 region encompassing amino acids 226-239 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen-binding domain binds to an epitope region encompassing amino acids 16-23 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen-binding domain binds to an epitope region encompassing amino acids 15-23 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen-binding domain binds to an epitope region encompassing amino acids 16-25 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 10-17 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 5-17 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 85-95 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 157-164 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 157-167 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 230-237 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 229-237 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 230-243 of SEQ ID NO: 97. In certain embodiments, the extracellular antigen binding domain binds to an epitope region comprising amino acids 227-237 of SEQ ID NO: 97.

[0030] In certain embodiments, the extracellular antigen binding domain binds to one, two, or three epitope regions selected from the group consisting of an epitope region comprising amino acids 16-25 of SEQ ID NO: 97, an epitope region comprising amino acids 157-164 of SEQ ID NO: 97, and an epitope region comprising amino acids 229-237 of SEQ ID NO: 97. For example, the extracellular antigen binding domain comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 57 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 58. For example, the extracellular antigen binding domain comprises a V chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 208. H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 209 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 210 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 211 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 212 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 213 L CDR3.

[0031] In certain embodiments, the extracellular antigen binding domain binds to one, two, or three epitope regions selected from the group consisting of an epitope region comprising amino acids 5-17 of SEQ ID NO: 97, an epitope region comprising amino acids 85-95 of SEQ ID NO: 97, and an epitope region comprising amino acids 157-164 of SEQ ID NO: 97. For example, the extracellular antigen binding domain comprises a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 61 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 62. For example, the extracellular antigen binding domain comprises a V chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 214. H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 215 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 216 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 217 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 218 LCDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 219 L CDR3.

[0032] In certain embodiments, the extracellular antigen binding domain binds to one or two epitope regions selected from the epitope region comprising amino acids 15-23 of SEQ ID NO: 97 and the epitope region comprising amino acids 230-243 of SEQ ID NO: 97. For example, the extracellular antigen binding domain comprises: a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 65 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 66. For example, the extracellular antigen binding domain comprises: a V chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 220. H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 221 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 222 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 223 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 224 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 225 L CDR3.

[0033] In certain embodiments, the extracellular antigen binding domain binds to one, two, or three epitope regions selected from the group consisting of an epitope region comprising amino acids 10-17 of SEQ ID NO: 97, an epitope region comprising amino acids 157-167 of SEQ ID NO: 97, and an epitope region comprising amino acids 227-237 of SEQ ID NO: 97. For example, the extracellular antigen binding domain comprises: a heavy chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 69 and a light chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 70. For example, the extracellular antigen binding domain comprises: a V chain variable region comprising amino acids having the sequence set forth in SEQ ID NO: 226. H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 227 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 228 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 229 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 230L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 231 L CDR3.

[0034] According to the subject matter disclosed in the present invention, the extracellular antigen binding domain is covalently bound to the membrane spaning domain.The extracellular antigen binding domain may include a signal peptide covalently bound to the 5' end of the extracellular antigen binding domain.In certain embodiments, the membrane spaning domain of CAR includes CD8 polypeptides, CD28 polypeptides, CD3ζ polypeptides, CD4 polypeptides, 4-1BB polypeptides, OX40 polypeptides, ICOS polypeptides, CTLA-4 polypeptides, PD-1 polypeptides, LAG-3 polypeptides, 2B4 polypeptides, BTLA polypeptides, synthetic peptides (non-based on proteins associated with immune responses), or combinations thereof.In certain embodiments, the membrane spaning domain includes CD8 polypeptides.In certain embodiments, the membrane spaning domain includes CD28 polypeptides.

[0035] According to the subject matter disclosed herein, the intracellular domain comprises a CD3 ζ polypeptide. In certain embodiments, the intracellular domain further comprises at least one signaling region. In certain embodiments, the at least one signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, a PD-1 polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide (not based on a protein associated with an immune response), or a combination thereof. In certain embodiments, the signaling region is a costimulatory signaling region. In certain embodiments, the costimulatory signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combination thereof. In certain embodiments, the at least one costimulatory signaling region comprises a CD28 polypeptide. In certain embodiments, the at least one costimulatory signaling region comprises a 4-1BB polypeptide. In certain embodiments, the transmembrane domain comprises a CD28 polypeptide, the intracellular domain comprises a CD3 ζ polypeptide, and the costimulatory signaling domain comprises a CD28 polypeptide.

[0036] In certain embodiments, the CAR is recombinantly expressed. The CAR can be expressed by a vector. In certain embodiments, the vector is a gamma-retroviral vector.

[0037] The subject matter disclosed in the present invention also provides the immune response cell of separation comprising the above-mentioned CAR.In certain embodiments, the immune response cell separated is transduced with CAR, for example, CAR is constitutively expressed on the surface of the immune response cell.In certain embodiments, the immune response cell separated is further transduced with at least one costimulatory ligand so that the immune response cell expresses the at least one costimulatory ligand.In certain embodiments, at least one costimulatory ligand is selected from 4-1BBL, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, and combinations thereof.In certain embodiments, the immune response cell separated is further transduced with at least one cytokine so that the immune response cell secretes the at least one cytokine.In certain embodiments, at least the cytokine is selected from IL-2, IL-3, IL-6, IL-7, IL-11, IL-12, IL-15, IL-17, IL-21, and combinations thereof. In some embodiments, the isolated immune response cells are selected from T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTL), regulatory T cells, human embryonic stem cells, lymphoid progenitor cells, T cell precursor cells and pluripotent stem cells that can differentiate into lymphoid cells. In certain embodiments, the immune response cells are T cells.

[0038] The presently disclosed subject matter also provides nucleic acid molecules encoding the CAR disclosed herein, vectors comprising the nucleic acid molecules, and host cells expressing such nucleic acid molecules. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 397. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 398. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 399. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 400. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 401. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 402. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 403. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 406. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 407. In certain embodiments, the nucleic acid molecule comprises a nucleic acid having a sequence as shown in SEQ ID NO: 408. In certain embodiments, the vector is a γ-retroviral vector. In certain embodiments, the host cell is a T cell.

[0039] In addition, the subject matter disclosed in the present invention provides a method for using the above-mentioned immune response cells to reduce the tumor load of a subject. For example, the subject matter disclosed in the present invention provides a method for reducing the tumor load of a subject, wherein the method comprises administering an effective amount of the immune response cells disclosed in the present invention to the subject, thereby inducing tumor cell death in the subject. In certain embodiments, the method reduces the number of tumor cells. In another embodiment, the method reduces tumor size. In yet another embodiment, the method eradicates the tumor in the subject. In certain embodiments, the subject is a human. In certain embodiments, the immune response cells are T cells. In certain embodiments, the tumor is multiple myeloma or Waldenstrom's Macroglobulinemia. In certain embodiments, the tumor is multiple myeloma.

[0040] In addition, the presently disclosed subject matter provides methods for using the above-described immune response cells to increase or prolong the survival of a subject suffering from a neoplasia. For example, the presently disclosed subject matter provides methods for increasing or prolonging the survival of a subject suffering from a neoplasia, wherein the method comprises administering to the subject an effective amount of the immune response cells disclosed herein, thereby increasing or prolonging the survival of the subject. In certain embodiments, the neoplasia is multiple myeloma or Waldenstrom's macroglobulinemia. In certain embodiments, the neoplasia is multiple myeloma. In certain embodiments, the method reduces or eradicates the tumor burden in the subject.

[0041] The subject matter disclosed in the present invention also provides a method for producing immune response cells in conjunction with G- protein coupled receptors. In a non-limiting example, the method includes introducing a nucleic acid sequence encoding a chimeric antigen receptor (CAR) into immune response cells, and the chimeric antigen receptor (CAR) includes an extracellular antigen binding domain, a transmembrane domain, and an intracellular domain, wherein the extracellular antigen binding domain specifically binds to G- protein coupled receptors. In certain embodiments, G- protein coupled receptors are G- protein coupled receptor C family 5 group D members (GPRC5D). In a specific non-limiting embodiment, the extracellular antigen binding domain is scFv.

[0042] The presently disclosed subject matter also provides pharmaceutical compositions comprising an effective amount of the immune response cells disclosed herein and a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical compositions are used to treat a neoplasia. In certain embodiments, the neoplasia is multiple myeloma or Waldenstrom's macroglobulinemia. In certain embodiments, the neoplasia is multiple myeloma.

[0043] The presently disclosed subject matter also provides kits for treating neoplasia comprising the immune response cells disclosed herein. In certain embodiments, the kits further include written instructions for using the immune response cells to treat the neoplasia. In certain embodiments, the neoplasia is multiple myeloma or Waldenstrom's macroglobulinemia. In certain embodiments, the neoplasia is multiple myeloma.

[0044] 1. A chimeric antigen receptor (CAR), comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular domain, wherein the extracellular antigen-binding domain specifically binds to a G-protein coupled receptor.

[0045] 2. The CAR according to embodiment 1, wherein the G protein receptor is a G-protein coupled receptor C family group 5 D member (GPRC5D).

[0046] 3. The CAR according to embodiment 2, wherein the extracellular antigen binding domain of the CAR is at about 1×10 -9 M is about 3×10 -6 The binding affinity (K d ) binds to GPRC5D.

[0047] 4. The CAR of embodiment 1, wherein the extracellular antigen binding domain is a single-chain variable fragment (scFv).

[0048] 5. The CAR of embodiment 4, wherein the extracellular antigen binding domain is a murine scFv.

[0049] 6. The CAR of embodiment 4, wherein the extracellular antigen binding domain is a human scFv.

[0050] 7. The CAR of embodiment 1, wherein the extracellular antigen binding domain is an optionally cross-linked Fab.

[0051] 8. The CAR of embodiment 1, wherein the extracellular antigen binding domain is F(ab)2.

[0052] 9. According to the CAR according to any one of embodiments 2-8, one or more of the scFv, Fab and F(ab)2 are contained in a fusion protein with a heterologous sequence to form the extracellular antigen binding domain.

[0053] 10. A CAR according to any one of embodiments 1-9, wherein the extracellular antigen binding domain comprises a heavy chain variable region, which contains an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to an amino acid sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374 and 386.

[0054] 11. A CAR according to any one of embodiments 1-10, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374 and 386 and conservative modifications thereof.

[0055] 12. A CAR according to any one of embodiments 1-11, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374 and 386.

[0056] 13. The CAR of embodiment 12, wherein the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:53.

[0057] 14. The CAR of embodiment 12, wherein the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO:57.

[0058] 15. The CAR of embodiment 12, wherein the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 61.

[0059] 16. The CAR of embodiment 12, wherein the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 65.

[0060] 17. The CAR of embodiment 12, wherein the extracellular antigen binding domain comprises a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 69.

[0061] 18. The CAR according to any one of embodiments 1-17, wherein the extracellular antigen binding domain comprises a light chain variable region comprising a light chain variable region selected from SEQ , about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence of ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375, and 387.

[0062] 19. A CAR according to any one of embodiments 1-18, wherein the extracellular antigen binding domain comprises: a light chain variable region comprising an amino acid sequence selected from SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387 and conservative modifications thereof.

[0063] 20. A CAR according to any one of embodiments 1-19, wherein the extracellular antigen binding domain comprises: a light chain variable region comprising amino acids with a sequence selected from SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387.

[0064] 21. The CAR of embodiment 20, wherein the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:54.

[0065] 22. The CAR of embodiment 20, wherein the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:58.

[0066] 23. The CAR of embodiment 20, wherein the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:62.

[0067] 24. The CAR of embodiment 20, wherein the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 66.

[0068] 25. The CAR of embodiment 20, wherein the extracellular antigen binding domain comprises a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO:70.

[0069] 26. The CAR according to any one of embodiments 1-25, wherein the extracellular antigen binding domain comprises: (a) a region containing a region selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374, and 386; and (b) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, , 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100,

[0070] 27. A CAR according to any one of embodiments 1-26, wherein the extracellular antigen binding domain comprises: (a) a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374 and 386 and conservative modifications thereof, and (b) a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: The amino acid sequences of NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387 and their conservatively modified light chain variable regions.

[0071] 28. A CAR according to any one of embodiments 1-27, wherein the extracellular antigen binding domain comprises: (a) a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374 and 386; and (b) a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375 and 387.

[0072] 29. The CAR of embodiment 28, wherein the extracellular antigen binding domain comprises:

[0073] (a) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 1 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 2;

[0074] (b) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 5 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 6;

[0075] (c) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 9 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 10;

[0076] (d) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 13 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 14;

[0077] (e) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 17 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 18;

[0078] (f) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 21 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 22;

[0079] (g) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 25 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 26;

[0080] (h) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 29 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 30;

[0081] (i) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 33 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 34;

[0082] (j) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 37 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 38;

[0083] (k) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 41 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 42;

[0084] (1) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 45 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 46;

[0085] (m) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 49 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 50;

[0086] (n) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 54;

[0087] (o) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 57 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 58;

[0088] (p) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 62;

[0089] (q) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 65 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 66;

[0090] (r) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 69 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 70;

[0091] (s) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 73 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 74;

[0092] (t) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 77 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 78;

[0093] (u) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 81 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 82;

[0094] (v) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 85 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 86;

[0095] (w) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 89 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 90;

[0096] (x) a heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 93 and a light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 94;

[0097] (y) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 302 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 303;

[0098] (z) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 314 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 315;

[0099] (aa) a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 326 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 327;

[0100] (ab) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 338 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 339;

[0101] (ac) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 350 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 351;

[0102] (ad) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 362 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 363;

[0103] (ae) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 374 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 375; or

[0104] (af) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 386 and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 387.

[0105] 30. A CAR according to embodiment 29, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 53; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 54.

[0106] 31. A CAR according to embodiment 29, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 57; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 58.

[0107] 32. The CAR of embodiment 29, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 61; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 62.

[0108] 33. A CAR according to embodiment 29, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 65; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 66.

[0109] 34. The CAR of embodiment 29, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 69; and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 70.

[0110] 35. A CAR according to any one of embodiments 1-34, wherein the extracellular antigen binding domain comprises a linker between the heavy chain variable region and the light chain variable region of the extracellular antigen binding domain.

[0111] 36. A CAR according to any one of embodiments 1-35, wherein the extracellular antigen binding domain comprises: (a) a heavy chain variable region CDR3 comprising an amino acid sequence selected from SEQ ID NO: 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222, 228, 234, 240, 246, 252, 258, 264, 306, 318, 330, 342, 354, 366, 378 and 390 and conservative modifications thereof; and (b) a heavy chain variable region CDR3 comprising an amino acid sequence selected from SEQ ID NO: The amino acid sequences of NO:129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 195, 201, 207, 213, 219, 225, 231, 237, 243, 249, 255, 261, 267, 309, 321, 333, 345, 357, 369, 381 and 393 and their conservatively modified light chain variable region CDR3.

[0112] 37. The CAR according to embodiment 36, wherein the extracellular antigen binding domain comprises: (a) a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305, 317, 329, 341, 353, 365, 377 and 389 and conservative modifications thereof; and (b) a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: The amino acid sequences of NO:128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 194, 200, 206, 212, 218, 224, 230, 236, 242, 248, 254, 260, 266, 308, 320, 332, 344, 356, 368, 380 and 392 and their conservatively modified light chain variable region CDR2.

[0113] 38. The CAR according to embodiment 36 or 37, wherein the extracellular antigen binding domain comprises: (a) a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 304, 316, 328, 340, 352, 364, 376 and 388, and conservative modifications thereof; and (b) a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: The amino acid sequences of NO:127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 193, 199, 205, 211, 217, 223, 229, 235, 241, 247, 253, 259, 265, 307, 319, 331, 343, 355, 367, 379 and 391 and their conservatively modified light chain variable region CDR1.

[0114] 39. A CAR according to any one of embodiments 1-38, wherein the extracellular antigen binding domain comprises: (a) a heavy chain variable region CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 304, 316, 328, 340, 352, 364, 376 and 388; (b) a heavy chain variable region CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: NO:125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305, 317, 329, 341, 353, 365, 377 and 389; (c) containing a heavy chain variable region CDR2 selected from the group consisting of SEQ (d) a heavy chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222, 228, 234, 240, 246, 252, 258, 264, 306, 318, 330, 342, 354, 366, 378, and 390; NO:127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 193, 199, 205, 211, 217, 223, 229, 235, 241, 247, 253, 259, 265, 307, 319, 331, 343, 355, 367, 379 and 391; (e) a light chain variable region CDR1 comprising an amino acid sequence selected from SEQ ID the light chain variable region CDR2 of the amino acid sequence of NO: 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 194, 200, 206, 212, 218, 224, 230, 236, 242, 248, 254, 260, 266, 308, 320, 332, 344, 356, 368, 380 and 392;and (f) a light chain variable region CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 195, 201, 207, 213, 219, 225, 231, 237, 243, 249, 255, 261, 267, 309, 321, 333, 345, 357, 369, 381, and 393;

[0115] 40. The CAR of any one of embodiments 1-39, wherein the extracellular antigen binding domain comprises:

[0116] (a) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 124 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 125 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 126 or a conservative modification thereof;

[0117] (b) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 130 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 131 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 132 or a conservative modification thereof;

[0118] (c) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 136 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 137 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 138 or a conservative modification thereof;

[0119] (d) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 142 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 143 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 144 or a conservative modification thereof;

[0120] (e) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 148 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 149 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 150 or a conservative modification thereof;

[0121] (f) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 154 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 155 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 156 or a conservative modification thereof;

[0122] (g) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 160 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 161 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 162 or a conservative modification thereof;

[0123] (h) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 166 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 167 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a conservative modification thereof;

[0124] (i) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 172 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 173 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 174 or a conservative modification thereof;

[0125] (j) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 178 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 179 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 180 or a conservative modification thereof;

[0126] (k) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 184 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 185 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a conservative modification thereof;

[0127] (1) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 190 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 191 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 192 or a conservative modification thereof;

[0128] (m) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 196 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 197 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 198 or a conservative modification thereof;

[0129] (n) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 202 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 203 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 204 or a conservative modification thereof;

[0130] (o) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 208 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 209 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 210 or a conservative modification thereof;

[0131] (p) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 214 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 215 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 216 or a conservative modification thereof;

[0132] (q) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 220 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 221 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 222 or a conservative modification thereof;

[0133] (r) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 226 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 227 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 228 or a conservative modification thereof;

[0134] (s) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 232 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 233 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 234 or a conservative modification thereof;

[0135] (t) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 238 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 239 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 240 or a conservative modification thereof;

[0136] (u) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 244 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 245 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 246 or a conservative modification thereof;

[0137] (v) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 250 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 251 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 252 or a conservative modification thereof;

[0138] (w) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 256 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 257 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 258 or a conservative modification thereof;

[0139] (x) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 262 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 263 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 264 or a conservative modification thereof;

[0140] (y) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 304 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 305 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 306 or a conservative modification thereof;

[0141] (z) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 316 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 317 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 318 or a conservative modification thereof;

[0142] (aa) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 328 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 329 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 330 or a conservative modification thereof;

[0143] (ab) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 340 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 341 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 342 or a conservative modification thereof;

[0144] (ac) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 352 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 353 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 354 or a conservative modification thereof;

[0145] (ad) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 364 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 365 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 366 or a conservative modification thereof;

[0146] (ae) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 376 or a conservative modification thereof; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 377 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 378 or a conservative modification thereof; or

[0147] (af) a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 388 or a conservative modification thereof; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 389 or a conservative modification thereof; and a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 390 or a conservative modification thereof.

[0148] 41. A CAR according to embodiment 40, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 202; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 203; and a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 204.

[0149] 42. A CAR according to embodiment 40, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 208; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 209; and a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 210.

[0150] 43. A CAR according to embodiment 40, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 214; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 215; and a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 216.

[0151] 44. The CAR according to embodiment 40, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 220; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 221; and a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 222.

[0152] 45. The CAR according to embodiment 40, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 226; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 227; and a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 228;

[0153] 46. ​​The CAR of any one of embodiments 1-45, wherein the extracellular antigen binding domain comprises:

[0154] (a) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 127 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 129 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 130 or a conservative modification thereof;

[0155] (b) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 133 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 134 or a conservative modification thereof; and a light chain variable region CDR3 comprising SEQ ID NO: 135 or a conservative modification thereof;

[0156] (c) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 139 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 140 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 141 or a conservative modification thereof;

[0157] (d) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 145 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 146 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 147 or a conservative modification thereof;

[0158] (e) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 151 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 152 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 153 or a conservative modification thereof;

[0159] (f) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 157 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 158 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 159 or a conservative modification thereof;

[0160] (g) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 163 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 164 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 165 or a conservative modification thereof;

[0161] (h) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 169 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 170 or a conservative modification thereof; and a light chain variable region CDR3 comprising SEQ ID NO: 171 or a conservative modification thereof;

[0162] (i) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 175 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 176 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 177 or a conservative modification thereof;

[0163] (j) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 181 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 182 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a conservative modification thereof;

[0164] (k) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 187 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 188 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a conservative modification thereof;

[0165] (1) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 193 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 194 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 195 or a conservative modification thereof;

[0166] (m) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 199 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 200 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a conservative modification thereof;

[0167] (n) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 205 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 206 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 207 or a conservative modification thereof;

[0168] (o) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 211 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 212 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 213 or a conservative modification thereof;

[0169] (p) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 217 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 218 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 219 or a conservative modification thereof;

[0170] (q) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 223 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 224 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 225 or a conservative modification thereof;

[0171] (r) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 229 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 230 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 231 or a conservative modification thereof;

[0172] (s) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 235 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 236 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 237 or a conservative modification thereof;

[0173] (t) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 241 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 242 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 243 or a conservative modification thereof;

[0174] (u) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 247 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 248 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 249 or a conservative modification thereof;

[0175] (v) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 253 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 254 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 255 or a conservative modification thereof;

[0176] (w) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 259 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 260 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 261 or a conservative modification thereof;

[0177] (x) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 265 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 266 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 267 or a conservative modification thereof;

[0178] (y) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 307 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 308 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 309 or a conservative modification thereof;

[0179] (z) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 319 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 320 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 321 or a conservative modification thereof;

[0180] (aa) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 331 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 332 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 333 or a conservative modification thereof;

[0181] (ab) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 343 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 344 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 345 or a conservative modification thereof;

[0182] (ac) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 355 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 356 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 357 or a conservative modification thereof;

[0183] (ad) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 367 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 368 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 369 or a conservative modification thereof;

[0184] (ae) a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 379 or a conservative modification thereof; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 380 or a conservative modification thereof; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 381 or a conservative modification thereof; or

[0185] (af) a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 391 or a conservative modification thereof; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 392 or a conservative modification thereof; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 393 or a conservative modification thereof.

[0186] 47. A CAR according to embodiment 46, wherein the extracellular antigen binding domain comprises: a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 205; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 206; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 207.

[0187] 48. A CAR according to embodiment 46, wherein the extracellular antigen binding domain comprises: a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 211; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 212; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 213.

[0188] 49. A CAR according to embodiment 46, wherein the extracellular antigen binding domain comprises: a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 217; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 218; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 219.

[0189] 50. A CAR according to embodiment 46, wherein the extracellular antigen binding domain comprises: a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 223; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 224; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 225.

[0190] 51. A CAR according to embodiment 46, wherein the extracellular antigen binding domain comprises: a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 229; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 230; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 231.

[0191] 52. The CAR of any one of embodiments 1-51, wherein the extracellular antigen binding domain comprises:

[0192] (a) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 124; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 125; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 126; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 127; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 128; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 129;

[0193] (b) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 130; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 131; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 132; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 133; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 134; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 135;

[0194] (c) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 136; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 137; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 138; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 139; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 140; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 141;

[0195] (d) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 142; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 143; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 144; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 145; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 146; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 147;

[0196] (e) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 148; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 149; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 150; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 151; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 152; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 153;

[0197] (f) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 154; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 155; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 156; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 157; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 158; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 159;

[0198] (g) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 160; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 161; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 162; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 163; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 164; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 165;

[0199] (h) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 166; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 167; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 168; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 169; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 170; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 171;

[0200] (i) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 172; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 173; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 174; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 175; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 176; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 177;

[0201] (j) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 178; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 179; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 180; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 181; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 182; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 183;

[0202] (k) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 184; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 185; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 186; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 187; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 188; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 189;

[0203] (1) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 190; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 191; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 192; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 193; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 194; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 195;

[0204] (m) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 196; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 197; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 198; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 199; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 200; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 201;

[0205] (n) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 202; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 203; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 204; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 205; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 206; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 207;

[0206] (o) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 208; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 209; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 210; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 211; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 212; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 213;

[0207] (p) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 214; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 215; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 216; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 217; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 218; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 219;

[0208] (q) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 220; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 221; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 222; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 223; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 224; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 225;

[0209] (r) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 226; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 227; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 228; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 229; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 230; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 231;

[0210] (s) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 232; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 233; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 234; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 235; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 236; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 237;

[0211] (t) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 238; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 239; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 240; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 241; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 242; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 243;

[0212] (u) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 244; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 245; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 246; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 247; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 248; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 249;

[0213] (v) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 250; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 251; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 252; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 253; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 254; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 255;

[0214] (w) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 256; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 257; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 258; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 259; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 260; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 261;

[0215] (x) a heavy chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 262; a heavy chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 263; a heavy chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 264; a light chain variable region CDR1 comprising the amino acid sequence set forth in SEQ ID NO: 265; a light chain variable region CDR2 comprising the amino acid sequence set forth in SEQ ID NO: 266; and a light chain variable region CDR3 comprising the amino acid sequence set forth in SEQ ID NO: 267;

[0216] (y) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 304; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 305; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 306; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 307; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 308; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 309;

[0217] (z) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 316; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 317; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 318; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 319; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 320; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 321;

[0218] (aa) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 328; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 329; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 330; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 331; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 332; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 333;

[0219] (ab) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 340; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 341; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 342; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 343; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 344; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 345;

[0220] (ac) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 352; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 353; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 354; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 355; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 356; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 357;

[0221] (ad) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 364; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 365; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 366; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 367; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 368; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 369;

[0222] (ae) a heavy chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 376; a heavy chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 377; a heavy chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 378; a light chain variable region CDR1 comprising the amino acid sequence of SEQ ID NO: 379; a light chain variable region CDR2 comprising the amino acid sequence of SEQ ID NO: 380; and a light chain variable region CDR3 comprising the amino acid sequence of SEQ ID NO: 381; or

[0223] (af) a heavy chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 388; a heavy chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 389; a heavy chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 390; a light chain variable region CDR1 comprising the amino acid sequence shown in SEQ ID NO: 391; a light chain variable region CDR2 comprising the amino acid sequence shown in SEQ ID NO: 392; and a light chain variable region CDR3 comprising the amino acid sequence shown in SEQ ID NO: 393.

[0224] 53. The CAR according to embodiment 52, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 202; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 203; a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 204; a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 205; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 206; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 207.

[0225] 54. The CAR according to embodiment 52, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 208; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 209; a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 210; a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 211; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 212; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 213.

[0226] 55. The CAR according to embodiment 52, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 214; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 215; a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 216; a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 217; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 218; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 219.

[0227] 56. The CAR according to embodiment 52, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 220; a heavy chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 221; a heavy chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 222; a light chain variable region CDR1 comprising an amino acid sequence as shown in SEQ ID NO: 223; a light chain variable region CDR2 comprising an amino acid sequence as shown in SEQ ID NO: 224; and a light chain variable region CDR3 comprising an amino acid sequence as shown in SEQ ID NO: 225.

[0228] 57. The CAR according to embodiment 52, wherein the extracellular antigen binding domain comprises: a heavy chain variable region CDR1 comprising amino acids as shown in SEQ ID NO: 226; a heavy chain variable region CDR2 comprising amino acids as shown in SEQ ID NO: 227; a heavy chain variable region CDR3 comprising amino acids as shown in SEQ ID NO: 228; a light chain variable region CDR1 comprising amino acids as shown in SEQ ID NO: 229; a light chain variable region CDR2 comprising amino acids as shown in SEQ ID NO: 230; and a light chain variable region CDR3 comprising amino acids as shown in SEQ ID NO: 231.

[0229] 58. A CAR according to any one of embodiments 1-57, wherein the extracellular antigen binding domain comprises a signal peptide covalently bound to the 5' end of the extracellular antigen binding domain.

[0230] 59. A CAR according to any one of embodiments 2-58, wherein the GPRC5D comprises the amino acid sequence shown in SEQ ID NO:97.

[0231] 60. The CAR according to embodiment 59, wherein the extracellular antigen binding domain binds to one, two, three or four epitope regions selected from: an epitope region in the N-terminal region comprising amino acids 1-27 of SEQ ID NO: 97, an epitope region in the ECL1 region comprising amino acids 85-93 of SEQ ID NO: 97, an epitope region in the ECL2 region comprising amino acids 145-167 of SEQ ID NO: 97, and an epitope region in the ECL3 region comprising amino acids 226-239 of SEQ ID NO: 97.

[0232] 61. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 16-23 of SEQ ID NO:97.

[0233] 62. The CAR of embodiment 61, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 15-23 of SEQ ID NO:97.

[0234] 63. The CAR of embodiment 61, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 16-25 of SEQ ID NO:97.

[0235] 64. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 10-17 of SEQ ID NO:97.

[0236] 65. The CAR of embodiment 64, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 5-17 of SEQ ID NO:97.

[0237] 66. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 85-95 of SEQ ID NO:97.

[0238] 67. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 157-164 of SEQ ID NO:97.

[0239] 68. The CAR of embodiment 67, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 157-167 of SEQ ID NO:97.

[0240] 69. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 230-237 of SEQ ID NO:97.

[0241] 70. The CAR of embodiment 69, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 229-237 of SEQ ID NO:97.

[0242] 71. The CAR of embodiment 69, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 230-243 of SEQ ID NO:97.

[0243] 72. The CAR of embodiment 69, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 227-237 of SEQ ID NO:97.

[0244] 73. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to one, two or three epitope regions selected from the group consisting of an epitope region comprising amino acids 16-25 of SEQ ID NO: 97, an epitope region comprising amino acids 157-164 of SEQ ID NO: 97, and an epitope region comprising amino acids 229-237 of SEQ ID NO: 97.

[0245] 74. A CAR according to embodiment 73, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 57 and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 58.

[0246] 75. The CAR according to embodiment 73 or embodiment 74, wherein the extracellular antigen binding domain comprises: a V comprising the amino acid sequence shown in SEQ ID NO: 208; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 209 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 210 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 211 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 212 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 213 L CDR3.

[0247] 76. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to one, two or three epitope regions selected from the group consisting of an epitope region comprising amino acids 5-17 of SEQ ID NO: 97, an epitope region comprising amino acids 85-95 of SEQ ID NO: 97, and an epitope region comprising amino acids 157-164 of SEQ ID NO: 97.

[0248] 77. A CAR according to embodiment 76, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 61 and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 62.

[0249] 78. The CAR according to embodiment 76 or embodiment 77, wherein the extracellular antigen binding domain comprises: a V comprising the amino acid sequence shown in SEQ ID NO: 214; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 215 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 216 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 217 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 218L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 219 L CDR3.

[0250] 79. A CAR according to embodiment 60, wherein the extracellular antigen binding domain binds to one or two epitope regions selected from the epitope region comprising amino acids 15-23 of SEQ ID NO:97 and the epitope region comprising amino acids 230-243 of SEQ ID NO:97.

[0251] 80. A CAR according to embodiment 79, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 65 and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 66.

[0252] 81. The CAR according to embodiment 79 or embodiment 80, wherein the extracellular antigen binding domain comprises: a V comprising the amino acid sequence shown in SEQ ID NO: 220; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 221 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 222 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 223 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 224 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 225 L CDR3.

[0253] 82. The CAR of embodiment 60, wherein the extracellular antigen binding domain binds to one, two or three epitope regions selected from the group consisting of an epitope region comprising amino acids 10-17 of SEQ ID NO: 97, an epitope region comprising amino acids 157-167 of SEQ ID NO: 97, and an epitope region comprising amino acids 227-237 of SEQ ID NO: 97.

[0254] 83. A CAR according to embodiment 82, wherein the extracellular antigen binding domain comprises: a heavy chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 69 and a light chain variable region comprising an amino acid sequence as shown in SEQ ID NO: 70.

[0255] 84. The CAR according to embodiment 82 or embodiment 83, wherein the extracellular antigen binding domain comprises: a V comprising the amino acid sequence shown in SEQ ID NO: 226; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 227 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 228 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 229 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 230 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 231 L CDR3.

[0256] 85. A CAR according to any one of embodiments 1-84, wherein the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide (not based on a protein associated with an immune response), or a combination thereof.

[0257] 86. The CAR of embodiment 85, wherein the transmembrane domain comprises a CD8 polypeptide.

[0258] 87. A CAR according to embodiment 85, wherein the transmembrane domain comprises a CD28 polypeptide.

[0259] 88. A CAR according to any one of embodiments 1-87, wherein the intracellular domain comprises a CD3ζ polypeptide.

[0260] 89. A CAR according to any one of embodiments 1-88, wherein the intracellular domain further comprises at least one signaling region.

[0261] 90. The CAR of embodiment 89, wherein at least one of the signaling regions comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, a PD-1 polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, a synthetic peptide (not based on a protein associated with an immune response), or a combination thereof.

[0262] 91. A CAR according to embodiment 89 or embodiment 90, wherein the signaling region is a co-stimulatory signaling region.

[0263] 92. The CAR of embodiment 91, wherein the at least one co-stimulatory signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combination thereof.

[0264] 93. A CAR according to embodiment 92, wherein at least one co-stimulatory signaling region comprises a CD28 polypeptide.

[0265] 94. The CAR of embodiment 92, wherein the at least one co-stimulatory signaling region comprises a 4-1BB polypeptide.

[0266] 95. A CAR according to any one of embodiments 89-93, wherein the transmembrane domain comprises a CD28 polypeptide, the intracellular domain comprises a CD3ζ polypeptide, and the signaling domain comprises a CD28 polypeptide.

[0267] 96. A CAR according to any one of embodiments 89-92 and 94, wherein the transmembrane domain comprises a CD8 polypeptide, the intracellular domain comprises a CD3ζ polypeptide, and the signaling domain comprises a 4-1BB polypeptide.

[0268] 97. The CAR of any one of embodiments 1-96, wherein the CAR is recombinantly expressed.

[0269] 98. The CAR of any one of embodiments 1-97, wherein the CAR is expressed by a vector.

[0270] 99. The CAR of embodiment 98, wherein the vector is a γ-retroviral vector.

[0271] 100. An isolated immune response cell comprising the CAR of any one of the preceding embodiments.

[0272] 101. An isolated immune response cell according to embodiment 100, wherein the immune response cell is transduced with the CAR.

[0273] 102. An isolated immune response cell according to embodiment 100 or 101, wherein the CAR is constitutively expressed on the surface of the immune response cell.

[0274] 103. An isolated immune response cell according to any one of embodiments 100-102, wherein the isolated immune response cell is further transduced with at least one co-stimulatory ligand such that the immune response cell expresses the at least one co-stimulatory ligand.

[0275] 104. The isolated immune response cell of embodiment 103, wherein the at least one co-stimulatory ligand is selected from 4-1BBL, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, and combinations thereof.

[0276] 105. The isolated immune response cell of any one of embodiments 100-104, wherein the isolated immune response cell is further transduced with at least one cytokine such that the immune response cell secretes the at least one cytokine.

[0277] 106. An isolated immune response cell according to embodiment 105, wherein the at least one cytokine is selected from IL-2, IL-3, IL-6, IL-7, IL-11, IL-12, IL-15, IL-17, IL-21, and combinations thereof.

[0278] 107. An isolated immune response cell according to any one of embodiments 100-106, wherein the immune response cell is selected from T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs), regulatory T cells, human embryonic stem cells, lymphoid progenitor cells, T cell precursor cells and pluripotent stem cells that can differentiate into lymphoid cells.

[0279] 108. An isolated immune response cell according to embodiment 107, wherein the immune response cell is a T cell.

[0280] 109. An isolated nucleic acid molecule encoding a chimeric antigen receptor (CAR) according to any one of embodiments 1-99.

[0281] 110. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO: 397.

[0282] 111. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:398.

[0283] 112. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:399.

[0284] 113. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO: 400.

[0285] 114. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:401.

[0286] 115. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:402.

[0287] 116. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:403.

[0288] 117. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:406.

[0289] 118. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:407.

[0290] 119. The isolated nucleic acid molecule of embodiment 109, comprising the nucleic acid having a sequence as shown in SEQ ID NO:408.

[0291] 120. A vector comprising the isolated nucleic acid molecule according to any one of embodiments 109-119.

[0292] 121. The vector of embodiment 120, wherein the vector is a γ-retroviral vector.

[0293] 122. A host cell expressing the nucleic acid molecule of any one of embodiments 109-119.

[0294] 123. A host cell according to embodiment 122, wherein the host cell is a T cell.

[0295] 124. A method of reducing tumor burden in a subject, comprising administering to the subject an effective amount of an immune response cell according to any one of embodiments 100-108, thereby inducing tumor cell death in the subject.

[0296] 125. The method of embodiment 124, wherein the method reduces the number of tumor cells.

[0297] 126. The method of embodiment 124, wherein the method reduces tumor size.

[0298] 127. The method of embodiment 124, wherein the method eradicates the tumor in the subject.

[0299] 128. The method of any one of embodiments 124-127, wherein the tumor is multiple myeloma or Waldenstrom's macroglobulinemia.

[0300] 129. The method of embodiment 128, wherein the tumor is multiple myeloma.

[0301] 130. The method of any one of embodiments 124-129, wherein the subject is human.

[0302] 131. A method according to any one of embodiments 124-130, wherein the immune response cell is a T cell.

[0303] 132. A method of increasing or prolonging the survival of a subject having a neoplasia, comprising administering to the subject an effective amount of an immune response cell according to any one of embodiments 100-108, thereby increasing or prolonging the survival of the subject.

[0304] 133. The method of embodiment 132, wherein the neoplasia is multiple myeloma or Waldenstrom's macroglobulinemia.

[0305] 134. The method of embodiment 133, wherein the neoplasia is multiple myeloma.

[0306] 135. The method of any one of embodiments 132-134, wherein the method reduces or eradicates the tumor burden in the subject.

[0307] 136. A method for preparing an immune response cell that binds to a G-protein coupled receptor, comprising introducing into the immune response cell

[0308] A nucleic acid sequence encoding a chimeric antigen receptor (CAR) comprising an extracellular antigen-binding domain, a transmembrane domain, and an intracellular domain, wherein the extracellular antigen-binding domain specifically binds to a G-protein coupled receptor.

[0309] 137. A method according to embodiment 136, wherein the G protein receptor is G-protein coupled receptor family C group 5 D member (GPRC5D).

[0310] 138. A pharmaceutical composition comprising an effective amount of the immune response cell according to any one of embodiments 100-108 and a pharmaceutically acceptable excipient.

[0311] 139. A pharmaceutical composition according to embodiment 138, wherein the pharmaceutical composition is used to treat neoplasia.

[0312] 140. The pharmaceutical composition of embodiment 139, wherein the neoplasia is multiple myeloma or Waldenstrom's macroglobulinemia.

[0313] 141. A pharmaceutical composition according to embodiment 140, wherein the neoplasia is multiple myeloma.

[0314] 142. A kit for treating multiple myeloma comprising the immune response cell according to any one of embodiments 100-108.

[0315] 143. A kit according to embodiment 142, wherein the kit further comprises written instructions for using the immune response cells to treat a subject suffering from neoplasia.

[0316] 144. The kit of embodiment 142 or 143, wherein the neoplasia is multiple myeloma or Waldenstrom's macroglobulinemia.

[0317] 145. The kit of embodiment 144, wherein the neoplasia is multiple myeloma. BRIEF DESCRIPTION OF THE DRAWINGS

[0318] The following detailed description may be understood in conjunction with the accompanying drawings, which are given by way of example and are not intended to limit the invention to the specific embodiments described.

[0319] Figure 1 A chimeric antigen receptor targeting a G-protein coupled receptor according to one non-limiting embodiment of the presently disclosed subject matter is shown.

[0320] Figure 2 Delineation of human GPRC5D expression in normal tissues and human cancer cell lines.

[0321] Figure 3 Depicts the expression of the disclosed GPRC5D CAR on human T cells.

[0322] Figure 4 The killing activity of GPRC5D disclosed in the present invention on 3T3 cells overexpressing GPRC5D is depicted.

[0323] Figure 5 The killing activity of GPRC5D disclosed in the present invention on human multiple myeloma cell lines was depicted.

[0324] Figure 6 A chimeric antigen receptor targeting GPRC5D according to one non-limiting embodiment of the presently disclosed subject matter is shown.

[0325] Figure 7 Depicted is a nucleic acid molecule encoding a CAR targeting GPRC5D according to one non-limiting embodiment of the presently disclosed subject matter.

[0326] Figure 8Depicted is a nucleic acid molecule encoding a CAR targeting GPRC5D according to one non-limiting embodiment of the presently disclosed subject matter.

[0327] Figure 9 Depicted is a nucleic acid molecule encoding a CAR targeting GPRC5D according to one non-limiting embodiment of the presently disclosed subject matter.

[0328] Figure 10 Delineation of the cytotoxicity of CAR T cells targeting GPRC5D against human multiple myeloma cell lines.

[0329] Figure 11 Depicting the induction of cytokine secretion by CAR T cells targeting GPRC5D.

[0330] Figure 12 Delineating the antitumor activity of CAR T cells targeting GPRC5D.

[0331] Figure 13A and Figure 13B Depicts the killing activity of CAR T cells targeting GPRC5D. (A) shows GFP at time 0 + (B) shows the percentage of tumor cell lines killed by GFP at 36 hours. + Percentage of tumor cell lines.

[0332] Figure 14 Illustration of the CLIPS technology. The CLIPS reaction occurs between the bromo groups of the CLIPS scaffold and the sulfhydryl side chains of cysteine. The reaction is rapid and specific under mild conditions. Using this ingenious chemistry, native protein sequences can be converted into CLIPS constructs with a range of structures. From left to right: two different single T2 loops, a T3 double loop, a conjugated T2+T3 loop, a stabilized beta sheet, and a stabilized alpha helix (Timmerman et al., J. Mol. Recognit. 2007;20:283-29).

[0333] Figure 15 Combinatorial CLIPS library screening is shown. A target protein containing discrete conformational epitopes (left) is converted into a matrix library (center). Combinatorial peptides are synthesized on proprietary microfilm and chemically converted into spatially defined CLIPS constructs (right).

[0334] Figure 16 Depiction of the T3 circularized CLIPS™ construct.

[0335] Figures 17A-17DIllustration of the heatmap technique. (A) Table of combined peptides, with two subsequences designated "Loop 1" and "Loop 2." (B) Data from A displayed as a matrix. (C) Color bar designation for the heatmap representation. (D) Heatmap visualization of data from A.

[0336] Figure 18 The intensity distribution recorded for ET150-2 is shown. A line is drawn from the start to the end residue of a peptide at the height of the signal recorded for a single peptide.

[0337] Figure 19 Shown is a heat map analysis of data recorded for ET150-5 under high stringency conditions.

[0338] Figure 20 The intensity distribution recorded by ET150-18 is shown.

[0339] Figure 21 Shown is the intensity distribution recorded by ET150-8.

[0340] Figure 22 Schematic diagram depicting a GPCR comprising seven transmembrane helices (TM) and three extracellular regions (ECL). The binding sites for each antibody are depicted with colored arrows.

[0341] Figure 23 A scatter plot analysis depicting all the data recorded for each sample. The slanted line is the statistical distribution.

[0342] Figure 24 Depicts FACS analysis of anti-GPRC5D antibodies.

[0343] Figure 25 Depicts FACS analysis of anti-GPRC5D antibodies.

[0344] Figure 26 Depicted is a nucleic acid molecule encoding a CAR targeting GPRC5D according to one non-limiting embodiment of the presently disclosed subject matter.

[0345] Figure 27 Depicted is a nucleic acid molecule encoding a CAR targeting GPRC5D according to one non-limiting embodiment of the presently disclosed subject matter. DETAILED DESCRIPTION

[0346] The subject matter disclosed herein generally provides a chimeric antigen receptor (CAR) targeting G- protein coupled receptors (e.g., G- protein coupled receptor C family 5 group D members (GPRC5D)). In a non-limiting example, CAR includes an extracellular antigen binding domain, a transmembrane domain, and an intracellular domain, wherein the extracellular antigen binding domain specifically binds to G- protein coupled receptors. The subject matter disclosed herein also provides immune response cells (e.g., T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTL), regulatory T cells, human embryonic stem cells, lymphoid progenitor cells, T cell precursor cells, and pluripotent stem cells that can differentiate into lymphoid cells) for expressing CAR targeting G- protein coupled receptors, and methods for treating cancers such as multiple myeloma using such immune response cells.

[0347] I. Definition

[0348] Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art to which the invention belongs. The following references provide general definitions of many of the terms used in the present invention for those skilled in the art: Singleton et al., Dictionary of Microbiology and Molecular Biology (Second Edition, 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, Fifth Edition, R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). Unless otherwise indicated, the following terms used herein have the meanings given below.

[0349] As used herein, the terms "about" or "approximately" mean within an acceptable error range for a particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, according to common practice in the art, "about" can mean within 3 or more standard deviations. Alternatively, "about" can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and even more preferably up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold of a value.

[0350] As used herein, the term "cell population" refers to a group of at least two cells expressing similar or different phenotypes. In a non-limiting example, a cell population can include at least about 10, at least about 100, at least about 200, at least about 300, at least about 400, at least about 500, at least about 600, at least about 700, at least about 800, at least about 900, or at least about 1000 cells expressing similar or different phenotypes.

[0351] The term "antibody" as used herein refers not only to complete antibody molecules, but also to antibody molecule fragments that retain the ability to bind to an immunogen. Such fragments are also well known in the art and are routinely used in vitro and in vivo. Therefore, the term "antibody" as used herein refers not only to complete immunoglobulin molecules, but also to the well-known active fragments F(ab')2 and Fab. F(ab')2 and Fab fragments, which lack the Fc fragment of a complete antibody, are cleared from the circulation more quickly and may have less non-specific tissue binding than complete antibodies (Wahl et al., J. Nucl. Med. 24: 316-325 (1983)). The antibodies of the present invention include complete natural antibodies, bispecific antibodies; chimeric antibodies; Fab, Fab', single-chain V region fragments (scFv), fusion polypeptides and unconventional antibodies.

[0352] As used herein, the term "single-chain variable fragment" or "scFv" is a fragment of a heavy chain (V H ) and light chain (V L ) of the variable region, which is covalently linked to form V H ::V L Heterodimer. Heavy chain (V H ) and light chain (V L ) are directly linked or connected through a linker encoding a peptide (e.g., 10, 15, 20, 25 amino acids) that connects V H The N-terminus of V L The C-terminus of the H The C-terminus of V L The N-terminus of the polypeptide is connected. The linker is generally rich in glycine for flexibility and serine or threonine for solubility. The linker can connect the heavy chain variable region and the light chain variable region of the extracellular antigen binding domain. Non-limiting examples of linkers are disclosed in Shen et al., Anal. Chem. 80 (6): 1910-1917 (2008) and WO 2014 / 087010, the entire contents of which are incorporated herein by reference. In certain embodiments, the linker is a G4S linker.

[0353] In a non-limiting example, the linker comprises amino acids having the sequence set forth in SEQ ID NO: 897, provided below.

[0354] GGGGSGGGGSGGGGS [SEQ ID NO: 284]. In certain embodiments, the nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 284 is shown in SEQ ID NO: 285, which is provided below:

[0355]

[0356] In another non-limiting example, the linker comprises amino acids having the sequence set forth in SEQ ID NO: 98, provided below.

[0357]

[0358] In certain embodiments, the nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:98 is shown in SEQ ID NO:99, which is provided below:

[0359]

[0360] Despite the removal of the constant region and the introduction of a linker, the scFv protein retains the specificity of the original immunoglobulin. H and V LNucleic acid expression of coding sequences is described in Huston et al. (Proc. Nat. Acad. Sci. USA, 85: 5879-5883, 1988). See also U.S. Patent Nos. 5,091,513, 5,132,405, and 4,956,778; and U.S. Patent Publication Nos. 20050196754 and 20050196754. Antagonistic scFvs with inhibitory activity have been described (see, e.g., Zhao et al., Hyrbidoma (Larchmt) 2008 27(6):455-51; Peter et al., J Cachexia Sarcopenia Muscle 2012 Aug 12; Shieh et al., J Imunol 2009 183(4):2277-85; Giomarelli et al., Thromb Haemost 2007 97(6):955-63; Fife et al., J Clin Invst 2006 116(8):2252-61; Brocks et al., Immunotechnology 1997 3(3):173-84; Moosmayer et al., Ther Immunol 1995 2(10):31-40). Agonistic scFvs with stimulatory activity have been described (see, e.g., Peter et al., J Biol ... Chern 2003 25278(38):36740-7; Xie et al., Nat Biotech 1997 15(8):768-71; Ledbetter et al., Crit Rev Immunol 1997 17(5-6):427-55; Ho et al., BioChim Biophys Acta 2003 1638(3):257-66).

[0361] As used herein, "F(ab)" refers to a fragment of an antibody structure that binds antigen but is monovalent and does not have an Fc portion, e.g., papain digestion of an antibody produces two F(ab) fragments and one Fc fragment (e.g., a heavy (H) chain constant region; the Fc region does not bind antigen).

[0362] As used herein, "F(ab')2" refers to an antibody fragment produced by pepsin digestion of an intact IgG antibody, wherein the fragment has two antigen-binding (ab') (divalent) regions, wherein each (ab') region comprises two separate amino acid chains, a portion of the H chain being linked to a light (L) chain by an SS bond for antigen binding, and wherein the remainder of the H chains are linked together. The "F(ab')2" fragment can be split into two separate Fab' fragments.

[0363] As used herein, the term "vector" refers to any genetic element, such as a plasmid, phage, transposon, cosmid, chromosome, virus, virion, etc., which is capable of replication when associated with appropriate control elements and which can transfer a gene sequence into a cell. Thus, the term includes cloning and expression vectors, as well as viral vectors and plasmid vectors.

[0364] As used herein, the term "expression vector" refers to a recombinant nucleic acid sequence, i.e., a recombinant DNA molecule, which contains a desired coding sequence and appropriate nucleic acid sequences necessary for expression of the operably linked coding sequence in a particular host organism. The nucleic acid sequences necessary for expression in prokaryotes typically include a promoter, an operator (optional), and a ribosome binding site, often accompanied by other sequences. Eukaryotic cells are known to utilize promoters, enhancers, and terminators, as well as polyadenylation signals.

[0365] As used herein, "CDR" is defined as the complementary determining region amino acid sequence of an antibody, which is the hypervariable region of the immunoglobulin heavy and light chains. See, for example, Kabat et al., Sequences of Proteins of Immunological Interest, 4th USDepartment of Health and Human Services, National Institutes of Health (1987). Typically, an antibody comprises three heavy chain and three light chain CDRs or CDR regions in the variable region. The CDRs provide the majority of contact residues for the antibody to bind to the antigen or epitope. In certain embodiments, the CDR regions are described using the Kabat system (Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242).

[0366] The term "affinity" as used herein refers to a measure of binding strength. Without being bound by theory, affinity depends on the tightness of the stereochemistry between the antibody binding site and the antigenic determinant, the size of the contact area between them, and the distribution of charged and hydrophobic groups. Affinity also includes the term "avidity," which refers to the intensity of the antigen-antibody bond after forming a reversible complex. Methods for calculating the affinity of an antibody for an antigen are known in the art and include calculating affinity using binding experiments. Antibody activity in functional tests (e.g., flow cytometry tests) also reflects antibody affinity. Functional tests (e.g., flow cytometry tests) can be used to phenotype antibodies and compare affinity.

[0367] Nucleic acid molecules useful in the methods of the present invention include any nucleic acid molecule encoding a polypeptide of the present invention or a fragment thereof. Such nucleic acid molecules need not be 100% identical to the endogenous nucleic acid sequence, but generally exhibit substantial identity. A polynucleotide having "substantial identity" to an endogenous sequence is generally capable of hybridizing to at least one strand of a double-stranded nucleic acid molecule. "Hybridization" refers to the pairing between complementary polynucleotide sequences (e.g., genes described herein) or portions thereof to form a double-stranded molecule under various stringent conditions. (See, e.g., Wahl, GM and SL Berger (1987) Methods Enzymol, 152: 399; Kimmel, AR (1987) Methods Enzymol, 152: 507).

[0368] For example, stringent salt concentrations are typically less than about 750 mM NaCl and 75 mM trisodium citrate, preferably less than about 500 mM NaCl and 50 mM trisodium citrate, more preferably less than about 250 mM NaCl and 25 mM trisodium citrate. Low stringency hybridization can be obtained in the absence of an organic solvent such as formamide, while high stringency hybridization can be obtained in the presence of at least about 35% formamide, more preferably at least about 50% formamide. Stringent temperature conditions typically include a temperature of at least about 30°C, more preferably at least about 37°C, most preferably at least about 42°C. It is well known to those skilled in the art to vary other parameters, such as hybridization time, detergent concentrations such as sodium dodecyl sulfate (SDS), and the inclusion or exclusion of vector DNA. Various stringent levels are achieved by combining these different conditions as needed. In a preferred embodiment, hybridization will occur at 30°C in 750 mM NaCl, 75 mM trisodium citrate, and 1% SDS. In a more preferred embodiment, hybridization will occur in 500 mM NaCl, 50 mM trisodium citrate, 1% SDS, 35% formamide, and 100 μg / ml denatured salmon sperm DNA (ssDNA) at 37° C. In a most preferred embodiment, hybridization will occur in 250 mM NaCl, 25 mM trisodium citrate, 1% SDS, 50% formamide, and 200 μg / ml ssDNA at 42° C. Useful variations on these conditions will be apparent to those skilled in the art.

[0369] For most applications, the washing step after hybridization will also vary in stringency. Washing stringency conditions can be defined by salt concentration and temperature. As mentioned above, washing stringency can be increased by reducing the salt concentration or by increasing the temperature. For example, the stringent salt concentration of the washing step is preferably less than about 30mM NaCl and 3mM trisodium citrate, and most preferably less than about 15mM NaCl and 1.5mM trisodium citrate. The stringent temperature conditions for the washing step typically include a temperature of at least about 25°C, more preferably at least about 42°C, and even more preferably at least about 68°C. In a preferred embodiment, the washing step will occur at 25°C in 30mM NaCl, 3mM trisodium citrate, and 0.1% SDS. In a more preferred embodiment, the washing step will occur at 42°C in 15mM NaCl, 1.5mM trisodium citrate, and 0.1% SDS. In a more preferred embodiment, the washing step will occur at 68°C in 15mM NaCl, 1.5mM trisodium citrate, and 0.1% SDS. Additional variations on these conditions will be readily apparent to those skilled in the art. Hybridization techniques are well known to those skilled in the art and are described, for example, in Benton and Davis (Science 196:180, 1977); Grunstein and Rogness (Proc. Natl. Acad. Sci., USA 72:3961, 1975); Ausubel et al. (Current Protocols in Molecular Biology, Wiley Interscience, New York, 2001); Berger and Kimmel (Guide to Molecular Cloning Techniques, 1987, Academic Press, New York); and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York.

[0370] "Substantial identity" refers to a polypeptide or nucleic acid molecule that exhibits at least 50% identity to a reference amino acid sequence (e.g., any of the amino acid sequences described herein) or nucleic acid sequence (e.g., any of the nucleic acid sequences described herein). Preferably, such sequences are at least 60%, more preferably 80% or 85%, more preferably 90%, 95% or even 99% identical at the amino acid level or nucleic acid to the sequence used for comparison.

[0371] Sequence identity is typically determined using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs). Such software matches identical or similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. In an exemplary method of determining the degree of identity, the BLAST program can be used, and probability scores between e-3 and e-100 indicate closely related sequences.

[0372] As used herein, the term "analog" refers to a structurally related polypeptide or nucleic acid molecule that has the function of a reference polypeptide or nucleic acid molecule.

[0373] As used herein, the term "ligand" refers to a molecule that binds to a receptor. In particular, a ligand binds to a receptor on another cell, allowing cell-to-cell recognition and / or interaction.

[0374] As used herein, the term "disease" refers to any condition or disorder that disrupts or interferes with the normal function of a cell, tissue, or organ. Examples of diseases include neoplasia or pathogen infection of cells.

[0375] The term "effective amount" as used herein refers to an amount sufficient to have a therapeutic effect. In certain embodiments, an "effective amount" is an amount sufficient to prevent, improve or inhibit the continued proliferation, growth or metastasis (eg, invasion or migration) of a neoplasia.

[0376] As used herein, the term "heterologous nucleic acid molecule or polypeptide" refers to a nucleic acid molecule (e.g., a cDNA, DNA, or RNA molecule) or polypeptide that is not normally present in a cell or in a sample obtained from a cell. The nucleic acid can be from another organism, or it can be, for example, an mRNA molecule that is not normally expressed in the cell or sample.

[0377] As used herein, the term "immune response cell" refers to a cell that plays a role in an immune response, or a progenitor cell thereof, or a progeny thereof.

[0378] As used herein, the term "modulate" means to change positively or negatively. Exemplary modulations include changes of about 1%, about 2%, about 5%, about 10%, about 25%, about 50%, about 75%, or about 100%.

[0379] As used herein, the term "increase" refers to a positive change of at least about 5%, including but not limited to a positive change of about 5%, about 10%, about 25%, about 30%, about 50%, about 75% or about 100%.

[0380] As used herein, the term "reduce" refers to a negative change of at least about 5%, including but not limited to a negative change of about 5%, about 10%, about 25%, about 30%, about 50%, about 75%, or about 100%.

[0381] As used herein, the term "isolated cell" refers to a cell that is separated from molecular and / or cellular components that naturally accompany the cell.

[0382] As used herein, the terms "isolated," "purified," or "biologically pure" refer to a substance that is free to varying degrees from the normally associated components found in its native state. "Isolated" refers to the degree of separation from the original source or environment. "Purified" refers to a higher degree of separation than isolated. A "purified" or "biologically pure" protein is sufficiently separated from other substances so that any impurities do not substantially affect the biological properties of the protein or cause other adverse consequences. That is, a nucleic acid or peptide of the invention is purified if it is substantially free of cellular material, viral material, or culture medium when produced by recombinant DNA technology, or substantially free of chemical precursors or other chemicals when chemically synthesized. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high performance liquid chromatography. The term "purified" can mean that a nucleic acid or protein produces essentially a single band in an electrophoretic gel. For proteins that can be modified (e.g., phosphorylated or glycosylated), different modifications can produce different isolated proteins that can be purified separately.

[0383] As used herein, the term "secreted" refers to the release of a polypeptide from a cell through the secretory pathway of the endoplasmic reticulum, the Golgi apparatus, and the release of the protein to the outside of the cell as vesicles that transiently fuse at the plasma membrane.

[0384] As used herein, the terms "specifically bind" or "specifically bind to" or "specifically target" refer to a polypeptide or fragment thereof that recognizes and binds to a biological molecule of interest (e.g., a polypeptide), but that does not substantially recognize and bind to other molecules in a sample, such as a biological sample that naturally includes a polypeptide of the invention.

[0385] The terms "treating" and "treatment" as used herein refer to clinical interventions that attempt to alter the disease process of the individual or cell being treated, and can be used for prevention or performed during a clinical pathological process. The therapeutic effects of treatment include, but are not limited to, preventing the occurrence or recurrence of the disease, alleviation of symptoms, reduction of any direct or indirect pathological consequences of the disease, prevention of metastasis, reduction of the rate of disease progression, alleviation or alleviation of the disease state, and alleviation or improvement of prognosis. By preventing the progression of the disease or disorder, treatment can prevent deterioration caused by the disorder in an affected or diagnosed subject or a subject suspected of having the disorder, and treatment can prevent the onset of the disorder or symptoms of the disorder in a subject at risk for or suspected of having the disorder.

[0386] As used herein, the term "subject" refers to any animal (eg, mammal), including but not limited to humans, non-human primates, rodents, etc. (eg, which is to be the recipient of a particular treatment, or from which cells are to be harvested).

[0387] II. G-protein coupled receptors

[0388] G protein-coupled receptors ("GPRs"), also known as seven-transmembrane domain receptors, 7TM receptors, seven-helix receptors, serpentine receptors, and G protein-linked receptors, constitute a large family of protein receptors that sense extracellular molecules and activate internal signal transduction pathways, ultimately activating cellular responses. GPCRs can be divided into six classes based on sequence homology and functional similarity: Class A (rhodopsin-like), Class B (secretin receptor family), Class C (metabolic glutamate / pheromones), Class D (fungal mating pheromone receptors), Class E (cyclic AMP receptors), and Class F (Frizzled / Smoothed). In certain embodiments, the GRP is a Class C GRP. In certain non-limiting embodiments, the Class C GRP is a G protein-coupled receptor C family 5 Group D member.

[0389] G protein-coupled receptor family C group 5 member D (GPRC5D) is an orphan receptor with no known ligand or function in humans. It is a member of the retinoic acid-inducible G protein-coupled receptor family. It is overexpressed in multiple myeloma (MM) cells and is not expressed or expressed at significantly lower levels in any other cell type, benign or malignant tumors, such as Figure 2 Several groups have compared gene expression analysis of primary MM cells with that of normal tissues. 1 or other hematological malignancies 2-4 This gene was identified as highly differentially expressed in comparison to controls. Higher mRNA expression has been shown to correlate with worse overall survival. 1 Surface staining of bone marrow aspirates from MM patients showed plasma cell-specific staining. 4To the best of the inventors' knowledge, this is the first time GPRC5D has been targeted by any therapy. In addition, to the best of the inventors' knowledge, this is the first time a CAR targeting any G protein-coupled receptor has been generated.

[0390] In certain non-limiting embodiments, the GPRC5D is human GPRC5D having the amino acid sequence shown in SEQ ID NO: 97, or a fragment thereof.

[0391] SEQ ID NO:97 is provided below:

[0392]

[0393] The N-terminal region of human GPRC5D has amino acids 1-27 of SEQ ID NO: 97. The extracellular loop 1 (ECL1) region of human GPRC5D has amino acids 85-93 of SEQ ID NO: 97. The extracellular loop 2 (ECL2) region of human GPRC5D has amino acids 145-167 of SEQ ID NO: 97. The extracellular loop 3 (ECL3) region of human GPRC5D has amino acids 226-239 of SEQ ID NO: 97.

[0394] III. Chimeric Antigen Receptor (CAR)

[0395] Chimeric antigen receptors (CARs) are engineered receptors that transfer or confer the specificity of interest to immune effector cells. CARs can be used to transfer the specificity of monoclonal antibodies to T cells by facilitating the transfer of their coding sequences via retroviral vectors.

[0396] There are three generations of CARs. “First generation” CARs typically consist of an extracellular antigen binding domain (e.g., a single chain variable fragment (scFv)) fused to a transmembrane domain, fused to the cytoplasmic / intracellular domain of the T cell receptor chain. “First generation” CARs typically have an intracellular domain from the CD3ζ chain, which is the primary transmitter of signals from the endogenous TCR. “First generation” CARs can provide de novo antigen recognition and activate CD4 through the CD3ζ chain signaling domain in a single fusion molecule. + and CD8 +T cells, rather than relying on HLA-mediated antigen presentation. "Second generation" CAR adds intracellular domains from various costimulatory molecules (such as CD28, 4-1BB, ICOS, OX40) to the cytoplasmic tail of CAR to provide additional signals to T cells. "Second generation" CAR includes those CARs that provide both costimulation (such as CD28 or 4-1BB) and activation (CD3ζ). Preclinical studies have shown that "second generation" CAR can improve the anti-tumor activity of T cells. For example, in patients with chronic lymphocytic leukemia (CLL) and acute lymphocytic leukemia (ALL), clinical trials targeting CD19 molecules have confirmed the strong efficacy of "second generation" CAR-modified T cells. "Third generation" CAR includes those CARs that provide multiple costimulation (such as CD28 and 4-1BB) and activation (CD3ζ).

[0397] According to the subject matter disclosed in the present invention, CAR comprises an extracellular antigen binding domain, a transmembrane domain and an intracellular domain, wherein the extracellular antigen binding domain binds to a G-protein coupled receptor. In certain embodiments, the G-protein coupled receptor is GPRC5D. In a specific non-limiting embodiment, the extracellular antigen binding domain is a scFv. In a specific non-limiting embodiment, the extracellular antigen binding domain is an optionally cross-linked Fab. In a specific non-limiting embodiment, the extracellular binding domain is F(ab)2. In a specific non-limiting embodiment, any of the foregoing molecules can be included in a fusion protein with a heterologous sequence to form an extracellular antigen binding domain.

[0398] In certain non-limiting embodiments, the extracellular antigen binding domain of the CAR disclosed herein has high binding specificity and high binding affinity to a G-protein coupled receptor (e.g., GPRC5D). For example, in such embodiments, the dissociation constant (K) of the extracellular antigen binding domain of the CAR (e.g., implemented as a scFv or its analogues) for binding to GPRC5D is D ) is about 3×10 -6 M or less. In certain embodiments, K D About 1×10 -6 M or smaller, approximately 1×10 -7 M or smaller, approximately 1×10 -8 M or smaller, or about 1×10 -9 M or smaller, approximately 1×10 -10 M or smaller, or about 1×10 -11 M or less. In certain embodiments, K D About 1×10 -8 M or less. In certain embodiments, K D About 1×10 -11M is about 3×10 -6 M, for example, about 1×10 -11 M is about 1×10 -10 M, about 1×10 -10 M is about 1×10 -9 M, about 1×10 -9 M is about 1×10 -8 M, about 1×10 -8 M is about 1×10 -7 M, or about 1×10 -7 M is about 1×10 -6 M, or about 1×10 -6 M is about 3×10 -6 M. In certain embodiments, K D About 1×10 -9 M is about 1×10 -8 M. In certain embodiments, K D About 1×10 -9 M is about 1.5×10 -9 M. In certain embodiments, K D About 1.2×10 -9 M. In certain embodiments, K D About 4×10 -9 M is about 5×10 -9 M. In certain embodiments, K D About 5×10 -9 M. In certain embodiments, K D About 4.8×10 -9 M. In certain embodiments, K D About 8×10 -9 M is about 9×10 -9 M. In certain embodiments, K D About 8×10 -9 M. In certain embodiments, K D About 8.1×10 -9 M.

[0399] The binding of the extracellular antigen binding domain of the CAR disclosed in the present invention (e.g., in the embodiment of scFv or its analogs) to the G-protein coupled receptor (e.g., GPRC5D) can be confirmed by, for example, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), FACS analysis, bioassay (e.g., growth inhibition) or Western Blot detection. Each of these tests is typically performed by using a labeling reagent (e.g., antibody or scFv) specific for the target complex to detect the presence of a specific target protein-antibody complex. For example, scFv can be radiolabeled and used for radioimmunoassay (RIA) (see, for example, Weintraub, B., Principles of Radioimmunoassays, Seventh Training Course on Radioligand Assay Techniques, The Endocrine Society, March 1986, which is incorporated herein by reference). Radioisotopes can be detected by, for example, using a γ counter or scintillation counter or by autoradiography. In certain embodiments, the extracellular antigen binding domain targeted by GPRC5D is labeled with a fluorescent marker. Non-limiting examples of fluorescent markers include green fluorescent protein (GFP), blue fluorescent proteins (e.g., EBFP, EBFP2, Azurite, and mKalama1), cyan fluorescent proteins (e.g., ECFP, Cerulean, and CyPet), and yellow fluorescent proteins (e.g., YFP, Citrine, Venus, and YPet). In certain embodiments, the human scFv targeting GPRC5D is labeled with GFP.

[0400] In certain embodiments, the extracellular antigen binding domain of CAR disclosed in the present invention includes a single-chain variable region fragment (scFv). In a specific embodiment, the extracellular antigen binding domain of CAR disclosed in the present invention includes a human scFv that specifically binds to human GPRC5D. In another specific embodiment, the extracellular antigen binding domain of CAR disclosed in the present invention includes a mouse scFv that specifically binds to human GPRC5D. In certain embodiments, identification is performed by screening a scFv phage library with cells expressing GPRC5D (e.g., 3T3 cells).

[0401] Extracellular antigen-binding domain of CAR

[0402] In certain embodiments, the extracellular antigen binding domain (e.g., scFv) comprises a heavy chain variable region comprising amino acids having a sequence selected from SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, and 93. The nucleic acid sequences encoding the amino acid sequences of SEQ ID NOs: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89 and 93 are SEQ ID NOs: 3, 7, 11, 15, 19, 23, 27, 31, 35, 39, 43, 47, 51, 55, 59, 63, 67, 71, 75, 79, 83, 87, 91 and 95, respectively. In some embodiments, the extracellular antigen binding domain (e.g., scFv) comprises a light chain variable region comprising amino acids having a sequence selected from SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, and 94. The nucleic acid sequences encoding the amino acid sequences of SEQ ID NOs: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90 and 94 are SEQ ID NOs: 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44, 48, 52, 56, 60, 64, 68, 72, 76, 80, 84, 88, 92 and 96, respectively. The sequences of SEQ ID NOs: 1-96 are described in Tables 1-24 below.

[0403] In certain embodiments, the extracellular antigen binding domain (eg, scFv) comprises a heavy chain variable region and a light chain variable region comprising amino acid sequences homologous to the amino acid sequences described herein and disclosed in Tables 1-24. For example, and not by way of limitation, the extracellular antigen binding domain (e.g., scFv) comprises a heavy chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93,

[0404] In certain embodiments, the extracellular antigen binding domain (e.g., scFv) comprises a light chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 2, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 303, 315, 327, 339, 351, 363, 375, and 387.

[0405] In certain embodiments, the extracellular antigen binding domain (eg, scFv) comprises (a) a fragment comprising a sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, 45, 49, 53, 57, 61, 65, 69, 73, 77, 81, 85, 89, 93, 302, 314, 326, 338, 350, 362, 374, and 386; and (b) a heavy chain variable region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, or about 99% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 5, 9, 13, 17, 21, 25, 29, 33, 37, 41, , 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100, 100,

[0406] The subject matter disclosed in the present invention also provides an extracellular antigen-binding domain (e.g., scFv) comprising a heavy chain variable region and a light chain variable region CDR, such as CDR1, CDR2, and CDR3, as disclosed in Tables 1-24 herein. CDR regions are described using the Kabat system (Kabat, EA, et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242). The subject matter disclosed in the present invention also provides an extracellular antigen-binding domain (e.g., scFv) comprising conservative modifications of antibody sequences disclosed herein. For example, and not by way of limitation, the extracellular antigen-binding domain (e.g., scFv) of the subject matter disclosed in the present invention comprises a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences and a light chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein one or more of these CDR sequences comprise a specified amino acid sequence disclosed herein or a conservative modification thereof, and wherein the extracellular antigen-binding domain retains the desired functional properties.

[0407] In certain embodiments, the presently disclosed subject matter provides an extracellular antigen binding domain (e.g., scFv) comprising a heavy chain variable region, wherein the heavy chain variable region comprises: (a) a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 304, 316, 328, 340, 352, 364, 376, and 388, and conservative modifications thereof; (b) a CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 NO:125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305, 317, 329, 341, 353, 365, 377 and 389 amino acid sequences and conservatively modified CDR2s thereof; and (c) comprising an amino acid sequence selected from the group consisting of SEQ ID The amino acid sequences of NO:126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222, 228, 234, 240, 246, 252, 258, 264, 306, 318, 330, 342, 354, 366, 378 and 390 and conservatively modified CDR3s thereof.

[0408] In certain embodiments, the extracellular antigen binding domain (e.g., scFv) comprises a light chain variable region, wherein the light chain variable region comprises: (a) a CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 193, 199, 205, 211, 217, 223, 229, 235, 241, 247, 253, 259, 265, 307, 319, 331, 343, 355, 367, 379, and 391, and conservative modifications thereof; (b) a CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 1 NO:128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 194, 200, 206, 212, 218, 224, 230, 236, 242, 248, 254, 260, 266, 308, 320, 332, 344, 356, 368, 380 and 392 amino acid sequences and conservatively modified CDR2s thereof; and (c) comprising an amino acid sequence selected from the group consisting of SEQ ID The amino acid sequences of NO:129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 195, 201, 207, 213, 219, 225, 231, 237, 243, 249, 255, 261, 267, 309, 321, 333, 345, 357, 369, 381 and 393 and their conservatively modified CDR3s.

[0409] The presently disclosed subject matter provides extracellular antigen binding domains (e.g., scFv) comprising a heavy chain variable region comprising a CDR1, CDR2, and CDR3 sequence and a light chain variable region comprising a CDR1, CDR2, and CDR3 sequence, wherein: (a) the heavy chain variable region CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 198, 204, 210, 216, 222, 228, 234, 240, 246, 252, 258, 264, 306, 318, 330, 342, 354, 366, 378, and 390, and conservative modifications thereof; and (b) the light chain variable region CDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: NO:129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 195, 201, 207, 213, 219, 225, 231, 237, 243, 249, 255, 261, 267, 309, 321, 333, 345, 357, 369, 381 and 393 amino acid sequences and conservative modifications thereof; wherein the extracellular antigen binding domain specifically binds to a GPRC5D polypeptide (e.g., a human GPRC5D polypeptide). In certain embodiments, the heavy chain variable region CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 197, 203, 209, 215, 221, 227, 233, 239, 245, 251, 257, 263, 305, 317, 329, 341, 353, 365, 377, and 389, and conservative modifications thereof; and (b) the light chain variable region CDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: NO:128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 194, 200, 206, 212, 218, 224, 230, 236, 242, 248, 254, 260, 266, 308, 320, 332, 344, 356, 368, 380 and 392 amino acid sequences and conservative modifications thereof; wherein the extracellular antigen binding domain specifically binds to a GPRC5D polypeptide (e.g., a human GPRC5D polypeptide).In certain embodiments, the heavy chain variable region CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 196, 202, 208, 214, 220, 226, 232, 238, 244, 250, 256, 262, 304, 316, 328, 340, 352, 364, 376, and 388, and conservative modifications thereof; and (b) the light chain variable region CDR1 comprises an amino acid sequence selected from the group consisting of SEQ ID NO: NO:127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 193, 199, 205, 211, 217, 223, 229, 235, 241, 247, 253, 259, 265, 307, 319, 331, 343, 355, 367, 379 and 391 amino acid sequences and conservative modifications thereof; wherein the extracellular antigen binding domain specifically binds to a GPRC5D polypeptide (e.g., a human GPRC5D polypeptide).

[0410] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 100 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-153 scFv (also referred to as "ET150-3 scFv").

[0411] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 1 and a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 2, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having the sequence shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 1. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 1. H, as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises amino acids having the sequence set forth in SEQ ID NO: 1, as shown in Table 1. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 2. L , as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 2, as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises: amino acids having a sequence as shown in SEQ ID NO: 1 V H and V comprising amino acids having a sequence shown in SEQ ID NO: 2 L , as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 124 or a conservative modification thereof H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 125 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 126 or a conservative modification thereof H CDR3, as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 127 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 128 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 129 or a conservative modification thereof L CDR3, as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 124 or a conservative modification thereof; H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 125 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 126 or a conservative modification thereof HCDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 127 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 128 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 129 or a conservative modification thereof L CDR3, as shown in Table 1. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 124; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 125 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 126 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 127 L CDR1, comprising amino acid V having the sequence shown in SEQ ID NO: 128 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 129 L CDR3.

[0412] Table 1

[0413]

[0414] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 101 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-166 scFv (also referred to as "ET150-16 scFv").

[0415] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 5 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 6, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V LThe extracellular antigen binding domain is selected from Table 2. In certain embodiments, the extracellular antigen binding domain is a human scFv. In certain embodiments, the extracellular antigen binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 5. H , as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence set forth in SEQ ID NO: 5, as shown in Table 2. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 6. L , as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 6, as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 5 H and V comprising amino acids having a sequence shown in SEQ ID NO: 6 L , as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 130 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 131 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 132 or a conservative modification thereof H CDR3, as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 133 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 134 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 135 or a conservative modification thereof LCDR3, as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 130 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 131 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 132 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 133 or a conservative modification thereof L CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 134 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 135 or a conservative modification thereof L CDR3, as shown in Table 2. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 130; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 131 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 132 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 133 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 134 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 135 L CDR3.

[0416] Table 2

[0417]

[0418] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 102 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-170 scFv (also referred to as "ET150-20 scFv").

[0419] In certain embodiments, the extracellular antigen-binding domain is an scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 9 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 10, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is an scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 3. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 9. H , as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence shown in SEQ ID NO: 9, as shown in Table 3. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 10. L , as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 10, as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 9 H and V comprising amino acids having a sequence shown in SEQ ID NO: 10 L , as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 136 or a conservative modification thereof. H CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 137 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 138 or a conservative modification thereof HCDR3, as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 139 or a conservative modification thereof. L CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 140 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 141 or a conservative modification thereof L CDR3, as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 136 or a conservative modification thereof. H CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 137 or a conservative modification thereof H CDR2, V comprising amino acids having a sequence as shown in SEQ ID NO: 138 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 139 or a conservative modification thereof L CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 140 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 141 or a conservative modification thereof L CDR3, as shown in Table 3. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 136; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 137 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 138 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 139 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 140 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 141 L CDR3.

[0420] Table 3

[0421]

[0422] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 103 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-171 scFv (also referred to as "ET150-21 scFv").

[0423] In certain embodiments, the extracellular antigen-binding domain is an scFv comprising: a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 13 and a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 14, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having the sequence shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is an scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 4. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 13. H , as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence shown in SEQ ID NO: 13, as shown in Table 4. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 14. L , as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 14, as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 13 H and V comprising amino acids having a sequence shown in SEQ ID NO: 14 L, as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 142 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 143 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 144 or a conservative modification thereof H CDR3, as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 145 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 146 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 147 or a conservative modification thereof L CDR3, as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 142 or a conservative modification thereof; H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 143 or a conservative modification thereof H CDR2, V comprising amino acids having a sequence as shown in SEQ ID NO: 144 or a conservative modification thereof H CDR3, V comprising amino acids having the sequence shown in SEQ ID NO: 145 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 146 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 147 or a conservative modification thereof L CDR3, as shown in Table 4. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 142; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 143 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 144 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 145 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 146 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 147L CDR3.

[0424] Table 4

[0425]

[0426] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 104 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-175 scFv (also referred to as "ET150-25 scFv").

[0427] In certain embodiments, the extracellular antigen-binding domain is an scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 17 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 18, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is an scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 5. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 17. H , as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises an amino acid sequence having a sequence as shown in SEQ ID NO: 17, as shown in Table 5. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 18. L , as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises V L, which comprises amino acids having a sequence as shown in SEQ ID NO: 18, as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 17 H and V comprising amino acids having a sequence shown in SEQ ID NO: 18 L , as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 148 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 149 or a conservative modification thereof H CDR2, and V comprising amino acids having the sequence shown in SEQ ID NO: 150 or a conservative modification thereof H CDR3, as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 151 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 152 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 153 or a conservative modification thereof L CDR3, as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 148 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 149 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 150 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 151 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 152 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 153 or a conservative modification thereof L CDR3, as shown in Table 5. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 148; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 149 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 150H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 151 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 152 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 153 L CDR3.

[0428] Table 5

[0429]

[0430] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 105 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-154 scFv (also referred to as "ET150-4 scFv").

[0431] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 21 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 22, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 6. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 21. H , as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises V H, which comprises the amino acids having the sequence set forth in SEQ ID NO: 21, as shown in Table 6. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 22. L , as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 22, as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 21 H and V comprising amino acids having a sequence shown in SEQ ID NO: 22 L , as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 154 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 155 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 156 or a conservative modification thereof H CDR3, as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 157 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 158 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 159 or a conservative modification thereof L CDR3, as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 154 or a conservative modification thereof; H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 155 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 156 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 157 or a conservative modification thereof LCDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 158 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 159 or a conservative modification thereof L CDR3, as shown in Table 6. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 154; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 155 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 156 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 157 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 158 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 159 L CDR3.

[0432] Table 6

[0433]

[0434]

[0435] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 106 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-156 scFv (also referred to as "ET150-6 scFv").

[0436] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 25 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 26, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V LThe extracellular antigen binding domain comprises a V region or CDR selected from Table 7. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 25. H , as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence set forth in SEQ ID NO: 25, as shown in Table 7. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 26. L , as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 26, as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 25 H and V comprising amino acids having a sequence shown in SEQ ID NO: 26 L , as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 160 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 161 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 162 or a conservative modification thereof H CDR3, as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 163 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 164 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 165 or a conservative modification thereof LCDR3, as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 160 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 161 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 162 or a conservative modification thereof H CDR3, V comprising amino acids having the sequence shown in SEQ ID NO: 163 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 164 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 165 or a conservative modification thereof L CDR3, as shown in Table 7. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 160; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 161 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 162 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 163 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 164 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 165 L CDR3.

[0437] Table 7

[0438]

[0439] In certain embodiments, the extracellular antigen-binding domain is a scFv that comprises the amino acid sequence of SEQ ID NO: 107 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-157 scFv (also referred to as "ET150-7 scFv").

[0440] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 29 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 30, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 8. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 29. H , as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence set forth in SEQ ID NO: 29, as shown in Table 8. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 30. L , as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 30, as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 29 H and V comprising amino acids having a sequence as shown in SEQ ID NO: 30 L , as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 166 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 167 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 168 or a conservative modification thereofH CDR3, as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 169 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 170 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 171 or a conservative modification thereof L CDR3, as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 166 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 167 or a conservative modification thereof H CDR2, V comprising amino acids having a sequence as shown in SEQ ID NO: 168 or a conservative modification thereof H CDR3, V comprising amino acids having the sequence shown in SEQ ID NO: 169 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 170 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 171 or a conservative modification thereof L CDR3, as shown in Table 8. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 166; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 167 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 168 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 169 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 170 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 171 L CDR3.

[0441] Table 8

[0442]

[0443] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 108 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-159 scFv (also referred to as "ET150-9 scFv").

[0444] In certain embodiments, the extracellular antigen-binding domain is an scFv comprising: a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 33 and a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 34, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having the sequence shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is an scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 9. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 33. H , as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises an amino acid sequence having a sequence as shown in SEQ ID NO: 33, as shown in Table 9. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 34. L , as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 34, as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 33 H and V comprising amino acids having a sequence as shown in SEQ ID NO: 34 L, as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 172 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 173 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 174 or a conservative modification thereof H CDR3, as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 175 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 176 or a conservative modification thereof L CDR2, and V comprising amino acids having the sequence shown in SEQ ID NO: 177 or conservative modifications thereof L CDR3, as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 172 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 173 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 174 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 175 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 176 or a conservative modification thereof L CDR2, and V comprising amino acids having the sequence shown in SEQ ID NO: 177 or conservative modifications thereof L CDR3, as shown in Table 9. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 172; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 173 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 174 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 175 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 176 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 177L CDR3.

[0445] Table 9

[0446]

[0447] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 109 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-160 scFv (also referred to as "ET150-10 scFv").

[0448] In certain embodiments, the extracellular antigen-binding domain is an scFv comprising: a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 37 and a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 38, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having the sequence shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is an scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 10. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 37. H , as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises an amino acid sequence having the sequence set forth in SEQ ID NO: 37, as shown in Table 10. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 38. L , as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises V L, which comprises amino acids having a sequence as shown in SEQ ID NO: 38, as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 37 H and V comprising amino acids having a sequence shown in SEQ ID NO: 38 L , as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 178 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 179 or a conservative modification thereof H CDR2, and V comprising amino acids having the sequence shown in SEQ ID NO: 180 or conservative modifications thereof H CDR3, as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 181 or a conservative modification thereof; L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 182 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 183 or a conservative modification thereof L CDR3, as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 178 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 179 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 180 or a conservative modification thereof H CDR3, V comprising amino acids having the sequence shown in SEQ ID NO: 181 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 182 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 183 or a conservative modification thereof L CDR3, as shown in Table 10. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 178; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 179 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 180H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 181 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 182 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 183 L CDR3.

[0449] Table 10

[0450]

[0451]

[0452] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 110 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-161 scFv (also referred to as "ET150-11 scFv").

[0453] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 41 and a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 42, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having the sequence shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 11. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 41. H , as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises V H, which comprises the amino acids having the sequence set forth in SEQ ID NO: 41, as shown in Table 11. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 42. L , as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 42, as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 41 H and V comprising amino acids having a sequence as shown in SEQ ID NO: 42 L , as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 184 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 185 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 186 or a conservative modification thereof H CDR3, as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 187 or a conservative modification thereof. L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 188 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 189 or a conservative modification thereof L CDR3, as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 184 or a conservative modification thereof; H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 185 or a conservative modification thereof H CDR2, V comprising amino acids having the sequence shown in SEQ ID NO: 186 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 187 or a conservative modification thereof LCDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 188 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 189 or a conservative modification thereof L CDR3, as shown in Table 11. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 184; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 185 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 186 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 187 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 188 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 189 L CDR3.

[0454] Table 11

[0455]

[0456]

[0457] In certain embodiments, the extracellular antigen-binding domain is a scFv that comprises the amino acid sequence of SEQ ID NO: 111 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-162 scFv (also referred to as "ET150-12 scFv").

[0458] In certain embodiments, the extracellular antigen-binding domain is an scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 45 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 46, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is an scFv-Fc fusion protein or a full-length human IgG, wherein V H and V LThe extracellular antigen binding domain comprises a V region or CDR selected from Table 12. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 45. H , as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence set forth in SEQ ID NO: 45, as shown in Table 12. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 46. L , as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 46, as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 45 H and V comprising amino acids having a sequence as shown in SEQ ID NO: 46 L , as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 190 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 191 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 192 or a conservative modification thereof H CDR3, as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 193 or a conservative modification thereof; L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 194 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 195 or a conservative modification thereof LCDR3, as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 190 or a conservative modification thereof; H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 191 or a conservative modification thereof H CDR2, V comprising amino acids having a sequence as shown in SEQ ID NO: 192 or a conservative modification thereof H CDR3, V comprising amino acids having the sequence shown in SEQ ID NO: 193 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 194 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 195 or a conservative modification thereof L CDR3, as shown in Table 12. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 190; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 191 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 192 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 193 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 194 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 195 L CDR3.

[0459] Table 12

[0460]

[0461] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising the amino acid sequence of SEQ ID NO: 112 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-163 scFv (also referred to as "ET150-13 scFv").

[0462] In certain embodiments, the extracellular antigen-binding domain is a scFv comprising: a heavy chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 49 and a light chain variable region comprising amino acids having a sequence as shown in SEQ ID NO: 50, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having a sequence as shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, wherein V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 13. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence of SEQ ID NO: 49. H , as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence set forth in SEQ ID NO: 49, as shown in Table 13. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 50. L , as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 50, as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 49 H and V comprising amino acids having a sequence shown in SEQ ID NO: 50 L , as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 196 or a conservative modification thereof. H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 197 or a conservative modification thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 198 or a conservative modification thereofH CDR3, as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 199 or a conservative modification thereof. L CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 200 or conservative modifications thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 201 or a conservative modification thereof L CDR3, as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 196 or a conservative modification thereof; H CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 197 or a conservative modification thereof H CDR2, V comprising amino acids having a sequence as shown in SEQ ID NO: 198 or a conservative modification thereof H CDR3, V comprising amino acids having a sequence as shown in SEQ ID NO: 199 or a conservative modification thereof L CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 200 or conservative modifications thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 201 or a conservative modification thereof L CDR3, as shown in Table 13. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 196; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 197 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 198 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 199 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 200 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 201 L CDR3.

[0463] Table 13

[0464]

[0465] In certain embodiments, the extracellular antigen-binding domain is a scFv that comprises the amino acid sequence of SEQ ID NO: 113 and specifically binds to a GPRC5D polypeptide (e.g., a GPRC5D polypeptide having the amino acid sequence of SEQ ID NO: 97, or a fragment thereof), and the scFv is designated as ET150-151 scFv (also referred to as "ET150-1 scFv").

[0466] In certain embodiments, the extracellular antigen-binding domain comprises: a heavy chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 53 and a light chain variable region comprising amino acids having the sequence shown in SEQ ID NO: 54, optionally with (iii) a linker sequence, such as a linker peptide, between the heavy chain variable region and the light chain variable region. In certain embodiments, the linker comprises amino acids having the sequence shown in SEQ ID NO: 98. In certain embodiments, the extracellular antigen-binding domain is a scFv-Fc fusion protein or a full-length human IgG, whose V H and V L The extracellular antigen binding domain comprises a V region or CDR selected from Table 14. In certain embodiments, the extracellular antigen binding domain comprises a V region comprising an amino acid sequence that is at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence shown in SEQ ID NO: 53. H , as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises V H , which comprises the amino acids having the sequence set forth in SEQ ID NO: 53, as shown in Table 14. In certain embodiments, the extracellular antigen-binding domain comprises a V comprising an amino acid sequence at least about 80%, about 81%, about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about 88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98% or about 99% homologous to the amino acid sequence set forth in SEQ ID NO: 54. L , as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises V L , which comprises amino acids having a sequence as shown in SEQ ID NO: 54, as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises: V comprising amino acids having a sequence as shown in SEQ ID NO: 53 H and V comprising amino acids having a sequence shown in SEQ ID NO: 54 L, as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 202 or a conservative modification thereof. H CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 203 or conservative modifications thereof H CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 204 or a conservative modification thereof H CDR3, as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 205 or a conservative modification thereof; L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 206 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 207 or a conservative modification thereof L CDR3, as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising amino acids having a sequence as shown in SEQ ID NO: 202 or a conservative modification thereof; H CDR1, V comprising amino acids having the sequence shown in SEQ ID NO: 203 or conservative modifications thereof H CDR2, V comprising amino acids having a sequence as shown in SEQ ID NO: 204 or a conservative modification thereof H CDR3, V comprising amino acids having the sequence shown in SEQ ID NO: 205 or a conservative modification thereof L CDR1, V comprising amino acids having a sequence as shown in SEQ ID NO: 206 or a conservative modification thereof L CDR2, and V comprising amino acids having a sequence as shown in SEQ ID NO: 207 or a conservative modification thereof L CDR3, as shown in Table 14. In certain embodiments, the extracellular antigen binding domain comprises: a V comprising an amino acid sequence having a sequence as shown in SEQ ID NO: 202; H CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 203 H CDR2, V comprising the amino acid sequence shown in SEQ ID NO: 204 H CDR3, V comprising the amino acid sequence shown in SEQ ID NO: 205 L CDR1, V comprising the amino acid sequence shown in SEQ ID NO: 206 L CDR2, and V comprising the amino acid sequence shown in SEQ ID NO: 207L CDR3.

[0467] Table 14

[0468]

[0469] In certain embodiments, the extracellular antigen-binding domain is a scFv that comprises the amino acid sequence of SEQ ID...

Claims

1. A chimeric antigen receptor (CAR), comprising an extracellular antigen binding domain that binds to G-protein coupled receptor C family 5 group D member (GPRC5D), a transmembrane domain and an intracellular domain, wherein the extracellular antigen binding domain comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises the CDR1, CDR2 and CDR3 sequences shown in SEQ ID NO: 208, SEQ ID NO: 209 and SEQ ID NO: 210, respectively, and the light chain variable region comprises the CDR1, CDR2 and CDR3 sequences shown in SEQ ID NO: 211, SEQ ID NO: 212 and SEQ ID NO: 213, respectively.

2. The CAR of claim 1, wherein the heavy chain variable region comprises an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO: 57; and the light chain variable region comprises an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO:

58.

3. The CAR according to claim 1, wherein the heavy chain variable region comprises the amino acid sequence shown in SEQ ID NO: 57, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:

58.

4. The CAR of claim 1, wherein the extracellular antigen binding domain comprises a linker between the heavy chain variable region and the light chain variable region of the extracellular antigen binding domain.

5. The CAR of claim 1, wherein the extracellular antigen binding domain is a single-chain variable fragment.

6. The CAR of claim 5, wherein the extracellular antigen binding domain is a human single-chain variable fragment.

7. The CAR according to claim 5, wherein the single-chain variable fragment comprises the amino acid sequence shown in SEQ ID NO:

114.

8. The CAR of claim 1, wherein the extracellular antigen binding domain is an optionally cross-linked Fab.

9. The CAR of claim 1, wherein the extracellular antigen binding domain is F(ab)2.

10. The CAR according to claim 1, wherein the extracellular antigen binding domain of the CAR is expressed in 1 × 10 -9 M to 3 × 10 -6 The dissociation constant (K d ) combined with GPRC5D.

11. The CAR of claim 1, wherein the extracellular antigen binding domain comprises a signal peptide covalently bound to the 5' end of the extracellular antigen binding domain.

12. The CAR according to claim 1, wherein the amino acid sequence of the GPRC5D is as shown in SEQ ID NO:

97.

13. The CAR according to claim 1, wherein the extracellular antigen binding domain binds to one or two epitope regions selected from: an epitope region within the N-terminal region of GPRC5D comprising amino acids 1-27 of SEQ ID NO: 97, and an epitope region within the ECL3 region of GPRC5D comprising amino acids 226-239 of SEQ ID NO:

97.

14. The CAR of claim 1, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 16-25 of SEQ ID NO:

97.

15. The CAR of claim 1, wherein the extracellular antigen binding domain binds to an epitope region comprising amino acids 229-237 of SEQ ID NO:

97.

16. The CAR of claim 1, wherein the extracellular antigen binding domain binds to one or two epitope regions selected from: an epitope region comprising amino acids 16-25 of SEQ ID NO:97 and an epitope region comprising amino acids 229-237 of SEQ ID NO:

97.

17. The CAR according to claim 1, wherein the extracellular antigen binding domain binds to a non-continuous epitope consisting of peptide segments 16-25 of SEQ ID NO: 97, 157-164 of SEQ ID NO: 97, and 229-237 of SEQ ID NO:

97.

18. The CAR of claim 1, wherein the transmembrane domain comprises a CD8 polypeptide, a CD28 polypeptide, a CD3ζ polypeptide, a CD4 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a CTLA-4 polypeptide, a PD-1 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, or a combination thereof.

19. The CAR of claim 1, wherein the transmembrane domain comprises a CD8 polypeptide.

20. The CAR of claim 1, wherein the transmembrane domain comprises a CD28 polypeptide.

21. The CAR of claim 1, wherein the intracellular domain comprises a CD3ζ polypeptide.

22. The CAR of claim 1, wherein the intracellular domain further comprises at least one signaling region.

23. The CAR of claim 22, wherein the at least one signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, a PD-1 polypeptide, a CTLA-4 polypeptide, a LAG-3 polypeptide, a 2B4 polypeptide, a BTLA polypeptide, or a combination thereof.

24. The CAR of claim 22, wherein the signaling region is a co-stimulatory signaling region.

25. The CAR of claim 24, wherein the co-stimulatory signaling region comprises a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, or a combination thereof.

26. The CAR of claim 25, wherein the co-stimulatory signaling region comprises a CD28 polypeptide.

27. The CAR of claim 25, wherein the co-stimulatory signaling region comprises a 4-1BB polypeptide.

28. The CAR of claim 1, wherein the transmembrane domain comprises a CD28 polypeptide, and the intracellular domain comprises a CD3ζ polypeptide and at least one signaling region comprising a CD28 polypeptide.

29. The CAR of claim 1, wherein the transmembrane domain comprises a CD8 polypeptide, and the intracellular domain comprises a CD3ζ polypeptide and at least one signaling region comprising a 4-1BB polypeptide.

30. The CAR of claim 1, wherein the transmembrane domain comprises a CD28 polypeptide, and the intracellular domain comprises a CD3ζ polypeptide and at least one signaling region comprising a 4-1BB polypeptide.

31. The CAR of claim 1, wherein the CAR is recombinantly expressed.

32. The CAR of claim 1, wherein the CAR is expressed by a vector.

33. The CAR of claim 32, wherein the vector is a γ-retroviral vector.

34. The CAR of claim 32, wherein the vector is a lentiviral vector.

35. An immune response cell comprising the CAR of any one of claims 1-34, wherein the immune response cell is selected from T cells, natural killer (NK) cells, cytotoxic T lymphocytes (CTLs), regulatory T cells, lymphoid progenitor cells, T cell precursor cells, and pluripotent stem cells that can differentiate into lymphoid cells.

36. The immune response cell of claim 35, wherein the immune response cell is transduced with the CAR.

37. The immune response cell of claim 35, wherein the CAR is constitutively expressed on the surface of the immune response cell.

38. The immune response cell of claim 35, wherein the immune response cell is further transduced with at least one co-stimulatory ligand such that the immune response cell expresses the at least one co-stimulatory ligand.

39. The immune response cell of claim 38, wherein the at least one co-stimulatory ligand is selected from 4-1BBL, CD80, CD86, CD70, OX40L, CD48, TNFRSF14, and combinations thereof.

40. The immune response cell of claim 35, wherein the immune response cell is further transduced with at least one cytokine such that the immune response cell secretes the at least one cytokine.

41. The immune response cell of claim 40, wherein the at least one cytokine is selected from IL-2, IL-3, IL-6, IL-7, IL-11, IL-12, IL-15, IL-17, IL-21, and combinations thereof.

42. The immune response cell of claim 35, wherein the immune response cell is a T cell.

43. A nucleic acid molecule encoding the chimeric antigen receptor (CAR) according to any one of claims 1-34.

44. According to the nucleic acid molecule according to claim 43, the nucleic acid sequence encoding the CAR is shown in SEQ ID NO:

397.

45. According to the nucleic acid molecule according to claim 43, the nucleic acid sequence encoding the CAR is shown in SEQ ID NO:

407.

46. ​​A vector comprising the nucleic acid molecule according to any one of claims 43-45.

47. The vector of claim 46, wherein the vector is a γ-retroviral vector.

48. The vector of claim 46, wherein the vector is a lentiviral vector.

49. A host cell expressing a nucleic acid molecule according to any one of claims 43-45.

50. The host cell of claim 49, wherein the host cell is a T cell.

51. Use of the immune response cell according to any one of claims 35-42 in the preparation of a medicament for treating multiple myeloma in a subject.

52. The use according to claim 51, wherein the subject is a human.

53. The use according to claim 51, wherein the immune response cells are T cells.

54. Use of an immune response cell according to any one of claims 35-42 in the preparation of a medicament for increasing or prolonging the survival of a subject with multiple myeloma.

55. A pharmaceutical composition comprising the immune response cell according to any one of claims 35-42 and a pharmaceutically acceptable excipient.

56. A kit for treating a tumor, comprising the immune response cell according to any one of claims 35-42.

57. The kit of claim 56, wherein the kit further comprises written instructions for using the immune response cells for treating a subject having a tumor.

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