Anti- DLL3 chimeric antigens receptors and uses thereof

Abstract

Provided are anti- DLL3 chimeric antigens receptors (CARs), DLL3 binding proteins, and uses of such CARs or DLL3 binding proteins in the treatment of DLL3 associated disorders (e.g., small cell lung carcinoma).SOLUTION: Provided is a chimeric antigen receptor (CAR) comprising a DLL3 binding domain, wherein the DLL3 binding domain comprises or is derived from a single domain antibody (sdAb) or a single chain variable fragment (scFv).SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to chimeric antigen receptors (CARs) that target DLL3 and binding proteins specific for DLL3. The present invention also relates to nucleic acid sequences encoding the CARs or binding proteins, modified immune cells that express the CARs, and their use to treat DLL3-associated disorders. [Background technology]

[0002] Advances in cellular immunotherapy offer a promising approach for the treatment of various tumors. One such treatment involves genetically engineering immune cells, particularly T cells, to express chimeric antigen receptors (CARs) on their cell surface. Chimeric antigen receptors are proteins that transfer the specificity of monoclonal antibodies (mAbs) to the effector functions of T cells in a conventional manner. When CARs are expressed in T cells, CAR-modified T cells (CAR-T or CAR-T cells) acquire several properties, such as antigen-specific recognition, antitumor reactivity, and proliferation, and can therefore act as "living drugs" to eradicate targeted tumor cells. In principle, any antigen (e.g., a cell surface molecule) can be targeted by these CAR-T cells. CAR-T cell therapy may circumvent tolerance to self-antigens and provide treatment independent of the patient's MHC status. Recent studies have demonstrated remarkable clinical responses in leukemia and lymphoma patients using T cells engineered to express chimeric antigen receptors targeting CD19.

[0003] CARs are expressed as transmembrane proteins containing an antigen-specific binding site, a transmembrane region, and a signaling cytoplasmic domain (e.g., CD3ζ chain). The antigen-specific binding site is usually a single-chain variable fragment (scFv) derived from a monoclonal antibody, consisting of a heavy chain and a light chain connected by a flexible linker. Recently, CAR constructs have incorporated additional cytoplasmic domains from costimulatory molecules such as CD28 or 4-1BB to enhance T cell survival in vivo. CARs have also undergone other genetic modifications, such as the addition of cytokine genes or genes to circumvent immunosuppressive mechanisms at tumor sites.

[0004] The DLL3 (Delta-like Ligand 3) protein has been found to be clinically associated with various proliferative disorders, including tumors exhibiting neuroendocrine characteristics, such as small cell lung cancer (SCLC). SCLC, which originates from neuroendocrine precursor cells, comprises approximately 15% of all lung cancers and has one of the lowest 5-year survival rates at 6% (Alvarado-Luna et al., 2016, Transl Lung Cancer Res 5:26-38; Siegel et al., 2017, CA Cancer J Clin 67:7-30). This is because it is highly aggressive, with approximately two-thirds of patients presenting with metastatic disease at the time of diagnosis, and is highly refractory to conventional treatments (e.g., platinum-based chemotherapy).

[0005] Improved therapeutic approaches for treating SCLC and other DLL3-expressing cancers are needed. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Alvarado-Luna et al.,2016,Transl Lung Cancer Res 5:26-38;Siegel et al.,2017,CA Cancer J Clin 67:7-30 Summary of the Invention

[0007] In one aspect, the present disclosure provides a chimeric antigen receptor (CAR) that targets DLL3 (anti-DLL3 CAR). The anti-DLL3 CAR comprises a DLL3-binding domain, which comprises or is derived from a single-domain antibody (sdAb) or a single-chain variable fragment (scFv).

[0008] In some embodiments, the sdAb comprises a polypeptide comprising a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81 or a variant thereof comprising up to about three amino acid substitutions in CDR1, a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162 or a variant thereof comprising up to about three amino acid substitutions in CDR2, and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243 or a variant thereof comprising up to about three amino acid substitutions in CDR3.

[0009] In some embodiments, the sdAb comprises a polypeptide comprising a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81, a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162, and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243, or a variant of the polypeptide comprising up to about three amino acid substitutions in CDR1, CDR2, and CDR3.

[0010] In some embodiments, the sdAb is: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. The present invention also includes a polypeptide comprising any one of the following:

[0011] In some embodiments, the sdAb is: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 The present invention also includes a polypeptide comprising any one of the following:

[0012] In some embodiments the sdAb is a camelid sdAb raised against human or rhesus DLL3.

[0013] In some embodiments, the sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274-354.

[0014] In some embodiments, the sdAb is humanized through CDR grafting.

[0015] In some embodiments, the humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355-367.

[0016] In some embodiments, the scFv comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO: 498 or 504 or a variant thereof comprising up to about three amino acid substitutions in CDR1, CDR2 set forth in SEQ ID NO: 499 or 505 or a variant thereof comprising up to about three amino acid substitutions in CDR2, and CDR3 set forth in SEQ ID NO: 500 or 506 or a variant thereof comprising up to about three amino acid substitutions in CDR3; and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO: 495 or 501 or a variant thereof comprising up to about three amino acid substitutions in CDR1, CDR2 set forth in SEQ ID NO: 496 or 502 or a variant thereof comprising up to about three amino acid substitutions in CDR2, and CDR3 set forth in SEQ ID NO: 497 or 503 or a variant thereof comprising up to about three amino acid substitutions in CDR3.

[0017] In some embodiments, the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO: 498, CDR2 set forth in SEQ ID NO: 499, and CDR3 set forth in SEQ ID NO: 500, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO: 495, CDR2 set forth in SEQ ID NO: 496, and CDR3 set forth in SEQ ID NO: 497; or the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO: 504, CDR2 set forth in SEQ ID NO: 505, and CDR3 set forth in SEQ ID NO: 506, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO: 501, CDR2 set forth in SEQ ID NO: 502, and CDR3 set forth in SEQ ID NO: 503.

[0018] In some embodiments, the VH domain of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 508, and the VL domain of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 507; or the VH domain of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 510, and the VL domain of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 509.

[0019] In some embodiments, the scFvs are obtained from a synthetic human Fab phage library.

[0020] In some embodiments, the DLL3 is human or rhesus DLL3.

[0021] In some embodiments, the anti-DLL3 CAR comprises, from N- to C-terminus, a signal peptide, a DLL3-binding domain, a hinge domain, a transmembrane domain, and an intracellular signaling domain.

[0022] In some embodiments, the intracellular signaling domain is derived from CD3zeta, FcRgamma, FcRbeta, CD3gamma, CD3delta, CD3epsilon, CD5, CD22, CD79a, CD79b, or CD66d.

[0023] In some embodiments, the intracellular signaling domain further comprises an intracellular costimulatory sequence.

[0024] In some embodiments, the intracellular costimulatory sequence is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD40, PD-1, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, TNFRSF9, TNFRSF4, TNFRSF8, CD40LG, ITGB2, KLRC2, TNFRSF18, TNFRSF14, HAVCR1, LGALS9, DAP10, DAP12, CD83, a ligand for CD83, and combinations thereof.

[0025] In some embodiments, the CAR comprises an amino acid sequence having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 476-484, 485-494, or 515-516.

[0026] In some embodiments, the DLL3 binding domain comprises two sdAbs linked to each other.

[0027] In some embodiments, each of the sdAbs independently comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:356 or SEQ ID NO:366.

[0028] In some embodiments, the CAR comprises an amino acid sequence having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 518-520.

[0029] In some embodiments, the CAR comprises the amino acid sequence of SEQ ID NO: 520.

[0030] In another aspect, the present disclosure provides a DLL3-binding protein comprising a single domain antibody (sdAb) portion that specifically binds to DLL3, wherein the sdAb portion comprises a polypeptide comprising: CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81 or a variant thereof comprising up to about three amino acid substitutions in CDR1; CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162 or a variant thereof comprising up to about three amino acid substitutions in CDR2; and CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243 or a variant thereof comprising up to about three amino acid substitutions in CDR3.

[0031] In some embodiments, the sdAb portion comprises a polypeptide comprising a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81, a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162, and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243, or a variant of the polypeptide comprising up to about three amino acid substitutions in CDR1, CDR2, and CDR3.

[0032] In some embodiments, the sdAb portion comprises: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. The present invention also includes a polypeptide comprising any one of the following:

[0033] In some embodiments, the sdAb portion comprises: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 The present invention also includes a polypeptide comprising any one of the following:

[0034] In some embodiments the sdAb moiety is a camelid sdAb raised against human or rhesus DLL3.

[0035] In some embodiments, the sdAb portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274-354.

[0036] In some embodiments, the sdAb portion is humanized through CDR grafting.

[0037] In some embodiments, the humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355-367.

[0038] In some embodiments, the DLL3 is human or rhesus DLL3.

[0039] In another aspect, the disclosure provides a DLL3 binding protein comprising a single chain variable fragment (scFv) portion that specifically binds to DLL3, wherein the scFv portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH domain of the scFv portion comprises a CDR1 set forth in SEQ ID NO: 498 or 504 or a variant thereof comprising up to about three amino acid substitutions in CDR1, a CDR2 set forth in SEQ ID NO: 499 or 505 or a variant thereof comprising up to about three amino acid substitutions in CDR2, and a CDR3 set forth in SEQ ID NO: 500 or 506. and the VL domain of the scFv portion comprises CDR1 set forth in SEQ ID NO: 495 or 501 or a variant thereof comprising up to about three amino acid substitutions in CDR1, CDR2 set forth in SEQ ID NO: 496 or 502 or a variant thereof comprising up to about three amino acid substitutions in CDR2, and CDR3 set forth in SEQ ID NO: 497 or 503 or a variant thereof comprising up to about three amino acid substitutions in CDR3.

[0040] In some embodiments, the scFv portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH domain of the scFv portion comprises CDR1 set forth in SEQ ID NO:498, CDR2 set forth in SEQ ID NO:499, and CDR3 set forth in SEQ ID NO:500, and the VL domain of the scFv portion comprises CDR1 set forth in SEQ ID NO:495, CDR2 set forth in SEQ ID NO:496, and CDR3 set forth in SEQ ID NO:497; or the VH domain of the scFv portion comprises CDR1 set forth in SEQ ID NO:504, CDR2 set forth in SEQ ID NO:505, and CDR3 set forth in SEQ ID NO:506, and the VL domain of the scFv portion comprises CDR1 set forth in SEQ ID NO:501, CDR2 set forth in SEQ ID NO:502, and CDR3 set forth in SEQ ID NO:503.

[0041] In some embodiments, the VH domain of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:508, and the VL domain of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:507; or the VH domain of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:510, and the VL domain of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:509.

[0042] In some embodiments, the scFv portion is obtained from a synthetic human Fab phage library.

[0043] In some embodiments, the DLL3 is human or rhesus DLL3.

[0044] In another aspect, the present disclosure provides an isolated nucleic acid molecule encoding an anti-DLL3 CAR or a DLL3 binding protein as described above.

[0045] In some embodiments, the isolated nucleic acid molecule comprises a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 368-448 that encodes a camelid single domain antibody (sdAb).

[0046] In some embodiments, the isolated nucleic acid molecule comprises a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 449-461 that encodes a humanized camelid sdAb.

[0047] In some embodiments, the isolated nucleic acid molecule comprises a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 511-514, encoding the VL or VH domain of a human scFv.

[0048] In some embodiments, the nucleic acid molecule further comprises a polynucleotide sequence encoding a chimeric switch receptor (CSR) or a dominant negative receptor (DNR).

[0049] In some embodiments, the nucleic acid molecule further comprises a polynucleotide sequence encoding a PD-1 dominant negative receptor (PD-1 DNR), a PD-1 chimeric switch receptor (PD-1 CSR), or a TGF-β dominant negative receptor (TGF-β DNR).

[0050] In some embodiments, the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:523.

[0051] In some embodiments, the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:524.

[0052] In some embodiments, the TGF-β DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:529.

[0053] In some embodiments, the polynucleotide sequence encoding the PD-1 DNR, PD-1 CSR, or TGF-β DNR is linked to the polynucleotide sequence encoding the CAR via a polynucleotide sequence encoding a 2A self-cleaving peptide.

[0054] In some embodiments, the 2A self-cleaving peptide is a T2A peptide or a P2A peptide.

[0055] In some embodiments, the nucleic acid molecule comprises, in the 5' to 3' direction, a polynucleotide sequence encoding a CAR, a polynucleotide sequence encoding a 2A self-cleaving peptide, and a polynucleotide sequence encoding a PD-1 DNR, a PD-1 CSR, or a TGF-β DNR.

[0056] In some embodiments, the nucleic acid molecule encodes a peptide having at least about 95% sequence identity to SEQ ID NO: 521 or 522, or a nucleic acid molecule encoding a peptide having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 525-528.

[0057] In another aspect, the disclosure provides an expression vector comprising an isolated nucleic acid molecule as described above.

[0058] In another aspect, the present disclosure provides a modified immune cell comprising an isolated nucleic acid molecule as described above.

[0059] In some embodiments, the engineered immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, a gamma delta T cell, an NKT cell, and a nature killer cell.

[0060] In another aspect, the present disclosure provides modified immune cells that express an anti-DLL3 CAR as described above.

[0061] In some embodiments, the modified immune cells also express CSR or DNR.

[0062] In some embodiments, the CSR is a PD-1 CSR and the DNR is a PD-1 DNR or a TGF-β DNR.

[0063] In some embodiments, the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:523.

[0064] In some embodiments, the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:524.

[0065] In some embodiments, the TGF-β DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:529.

[0066] In some embodiments, the CAR and CSR, or the CAR and DNR, are co-expressed via a 2A self-cleaving peptide.

[0067] In some embodiments, the 2A self-cleaving peptide is a T2A peptide or a P2A peptide.

[0068] In some embodiments, the engineered immune cells express a CAR and a PD-1 DNR and are stimulated by cells that express DLL3 and PD-L1.

[0069] In some embodiments, the engineered immune cells express a CAR and a PD-1 CSR and are stimulated by cells that express DLL3 and PD-L1.

[0070] In some embodiments, the engineered immune cells express a CAR and a TGF-β DNR, and are stimulated by cells expressing DLL3 in the presence of TGF-β.

[0071] In some embodiments, the engineered immune cells are selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and natural killer cells.

[0072] In another aspect, the present disclosure provides a pharmaceutical composition comprising an anti-DLL3 CAR, an isolated DLL3 binding protein, an expression vector, or a modified immune cell as described above, and a physiologically acceptable excipient.

[0073] In another aspect, the present disclosure provides a method for treating a DLL3-associated disorder in a subject, the method comprising administering to the subject a therapeutically effective amount of modified immune cells as described above, or a therapeutically effective amount of a pharmaceutical composition as described above.

[0074] In another aspect, the present disclosure provides use of an anti-DLL3 CAR, an isolated DLL3 binding protein, an expression vector, or a modified immune cell as described above for the preparation of a medicament for treating a DLL3-associated disorder.

[0075] In another aspect, the present disclosure provides a medicament for use in treating a DLL3-associated disorder, the medicament comprising an anti-DLL3 CAR, a DLL3 binding protein, an expression vector, or a modified immune cell as described above.

[0076] In some embodiments, the DLL3-associated disorder is a cancer selected from the group consisting of lung cancer, melanoma, breast cancer, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia, and lymphoma.

[0077] In some embodiments, the cancer expresses DLL3 and PD-L1.

[0078] In some embodiments, the cancer has a higher level of TGF-β expression compared to the corresponding normal tissue.

[0079] In some embodiments, the DLL3-associated disorder is small cell lung cancer. [The present invention 1001] A chimeric antigen receptor (CAR) comprising a DLL3-binding domain, wherein the DLL3-binding domain comprises or is derived from a single-domain antibody (sdAb) or a single-chain variable fragment (scFv). [The present invention 1002] 1001. A CAR of the present invention, wherein the sdAb comprises a polypeptide comprising: CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81 or a variant thereof comprising up to about three amino acid substitutions in the CDR1; CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162 or a variant thereof comprising up to about three amino acid substitutions in the CDR2; and CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243 or a variant thereof comprising up to about three amino acid substitutions in the CDR3. [The present invention 1003] The CAR of the present invention 1001 or 1002, wherein the sdAb comprises a polypeptide comprising CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81, CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162, and CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243, or a variant of the polypeptide comprising up to approximately three amino acid substitutions in the CDR1, the CDR2, and the CDR3. [The present invention 1004] the sdAb (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. A CAR of any one of 1001 to 1003, comprising a polypeptide comprising any one of the following: [The present invention 1005] the sdAb (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 A CAR of any one of 1001 to 1004, comprising a polypeptide comprising any one of the following: [The present invention 1006] The CAR of any of claims 1001 to 1005, wherein the sdAb is a camel sdAb raised against human or rhesus DLL3. [The present invention 1007] The CAR of any one of claims 1001 to 1006, wherein the sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274 to 354. [The present invention 1008] The CAR of any of claims 1001 to 1007, wherein the sdAb is humanized through CDR grafting. [The present invention 1009] The CAR of any one of claims 1001 to 1008, wherein the humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355 to 367. [The present invention 1010] The CAR of any of the present inventions 1001 to 1009, wherein the scFv comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH of the scFv comprises CDR1 shown in SEQ ID NO: 498 or 504 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, CDR2 shown in SEQ ID NO: 499 or 505 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and CDR3 shown in SEQ ID NO: 500 or 506 or a variant thereof comprising up to about three amino acid substitutions in the CDR3, and the VL of the scFv comprises CDR1 shown in SEQ ID NO: 495 or 501 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, CDR2 shown in SEQ ID NO: 496 or 502 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and CDR3 shown in SEQ ID NO: 497 or 503 or a variant thereof comprising up to about three amino acid substitutions in the CDR3. [The present invention 1011] A CAR of any of the present inventions 1001 to 1010, wherein the VH of the scFv comprises CDR1 shown in SEQ ID NO: 498, CDR2 shown in SEQ ID NO: 499, and CDR3 shown in SEQ ID NO: 500, and the VL of the scFv comprises CDR1 shown in SEQ ID NO: 495, CDR2 shown in SEQ ID NO: 496, and CDR3 shown in SEQ ID NO: 497; or the VH of the scFv comprises CDR1 shown in SEQ ID NO: 504, CDR2 shown in SEQ ID NO: 505, and CDR3 shown in SEQ ID NO: 506, and the VL of the scFv comprises CDR1 shown in SEQ ID NO: 501, CDR2 shown in SEQ ID NO: 502, and CDR3 shown in SEQ ID NO: 503. [The present invention 1012] A CAR of any of the present inventions 1001 to 1011, wherein the VH of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 508, and the VL of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 507; or the VH of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 510, and the VL of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 509. [The present invention 1013] The CAR of any of claims 1001 to 1012, wherein the scFv is obtained from a synthetic human Fab phage library. [The present invention 1014] The CAR of any of claims 1001 to 1013, wherein the DLL3 is human or rhesus DLL3. [The present invention 1015] The CAR of any of claims 1001 to 1014, comprising, from the N-terminus to the C-terminus, a signal peptide, the DLL3-binding domain, a hinge domain, a transmembrane domain, and an intracellular signaling domain. [The present invention 1016] The CAR of any one of claims 1001 to 1015, wherein the intracellular signaling domain is derived from CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, or CD66d. [The present invention 1017] The CAR of any one of claims 1001 to 1016, wherein the intracellular signaling domain further comprises an intracellular costimulatory sequence. [The present invention 1018] CAR of any of claims 1001 to 1017, wherein the intracellular costimulatory sequence is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD40, PD-1, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, TNFRSF9, TNFRSF4, TNFRSF8, CD40LG, ITGB2, KLRC2, TNFRSF18, TNFRSF14, HAVCR1, LGALS9, DAP10, DAP12, CD83, a ligand for CD83, and combinations thereof. [The present invention 1019] A CAR of any one of SEQ ID NOs: 476 to 484, 485 to 494, or 515 to 516, comprising an amino acid sequence having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 476 to 484, 485 to 494, or 515 to 516. [The present invention 1020] The CAR of any of claims 1001 to 1019, wherein the DLL3-binding domain comprises two sdAbs linked to each other. [The present invention 1021] The CAR of any of claims 1001 to 1020, wherein each of the sdAbs independently comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO: 356 or SEQ ID NO: 366. [The present invention 1022] A CAR of any of the present inventions 1001 to 1021, comprising an amino acid sequence having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 518 to 520. [The present invention 1023] A CAR of any one of 1001 to 1022 of the present invention, comprising the amino acid sequence of SEQ ID NO: 520. [The present invention 1024] A DLL3-binding protein comprising a single domain antibody (sdAb) portion that specifically binds to DLL3, wherein the sdAb portion comprises a polypeptide comprising: a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81 or a variant thereof comprising up to about three amino acid substitutions in the CDR1; a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162 or a variant thereof comprising up to about three amino acid substitutions in the CDR2; and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243 or a variant thereof comprising up to about three amino acid substitutions in the CDR3. [The present invention 1025] 1024. A DLL3-binding protein of the present invention, wherein the sdAb portion comprises a polypeptide comprising CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81, CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162, and CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243, or a variant of the polypeptide comprising up to about three amino acid substitutions in the CDR1, the CDR2, and the CDR3. [The present invention 1026] the sdAb portion (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. DLL3-binding protein of the present invention 1024 or 1025, comprising a polypeptide comprising any one of: [The present invention 1027] the sdAb portion (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 DLL3-binding protein according to any one of claims 1024 to 1026, comprising a polypeptide comprising any one of the following: [The present invention 1028] DLL3-binding protein of any one of claims 1024 to 1027, wherein the sdAb portion is a camel sdAb raised against human or rhesus DLL3. [The present invention 1029] DLL3-binding protein of any one of claims 1024 to 1028, wherein the sdAb portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274 to 354. [The present invention 1030] DLL3-binding protein of any of claims 1024 to 1029, wherein the sdAb portion is humanized through CDR grafting. [The present invention 1031] DLL3-binding protein of any of claims 1024 to 1030, wherein said humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355 to 367. [The present invention 1032] The DLL3-binding protein of any one of claims 1024 to 1031, wherein the DLL3 is human or rhesus DLL3. [The present invention 1033] 1. A DLL3 binding protein comprising a single chain variable fragment (scFv) portion that specifically binds to DLL3, wherein the scFv portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH of the scFv portion comprises a CDR1 set forth in SEQ ID NO: 498 or 504 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, a CDR2 set forth in SEQ ID NO: 499 or 505 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and a CDR3 set forth in SEQ ID NO: 500 or 506 or A DLL3-binding protein comprising a CDR1 set forth in SEQ ID NO: 495 or 501 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, a CDR2 set forth in SEQ ID NO: 496 or 502 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and a CDR3 set forth in SEQ ID NO: 497 or 503 or a variant thereof comprising up to about three amino acid substitutions in the CDR3. [The present invention 1034] A DLL3-binding protein of the present invention 1033, wherein the VH of the scFv portion comprises CDR1 set forth in SEQ ID NO: 498, CDR2 set forth in SEQ ID NO: 499, and CDR3 set forth in SEQ ID NO: 500, and the VL of the scFv portion comprises CDR1 set forth in SEQ ID NO: 495, CDR2 set forth in SEQ ID NO: 496, and CDR3 set forth in SEQ ID NO: 497; or the VH of the scFv portion comprises CDR1 set forth in SEQ ID NO: 504, CDR2 set forth in SEQ ID NO: 505, and CDR3 set forth in SEQ ID NO: 506, and the VL of the scFv portion comprises CDR1 set forth in SEQ ID NO: 501, CDR2 set forth in SEQ ID NO: 502, and CDR3 set forth in SEQ ID NO: 503. [This invention 1035] The DLL3 binding protein of the present invention 1033 or 1034, wherein the VH of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 508, and the VL of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 507; or the VH of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 510, and the VL of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 509. [The present invention 1036] DLL3-binding protein of any one of claims 1033 to 1035, wherein the scFv portion is obtained from a synthetic human Fab phage library. [This invention 1037] The DLL3-binding protein of any one of claims 1033 to 1036, wherein the DLL3 is human or rhesus DLL3. [The present invention 1038] An isolated nucleic acid molecule encoding a CAR of any one of the present inventions 1001 to 1023 or a DLL3-binding protein of any one of the present inventions 1024 to 1032 or 1033 to 1037. [This invention 1039] 1038. An isolated nucleic acid molecule of the present invention, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 368 to 448, encoding a camelid single domain antibody (sdAb). [The present invention 1040] 1038 or 1039, an isolated nucleic acid molecule of the present invention, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 449 to 461, encoding a humanized camelid sdAb. [The present invention 1041] The isolated nucleic acid molecule of any one of claims 1038 to 1040, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 511 to 514, encoding the VL or VH domain of human scFv. [The present invention 1042] The isolated nucleic acid molecule of any of claims 1038 to 1041, further comprising a polynucleotide sequence encoding a chimeric switch receptor (CSR) or a dominant negative receptor (DNR). [This invention 1043] The isolated nucleic acid molecule of any one of claims 1038 to 1042, further comprising a polynucleotide sequence encoding a PD-1 dominant negative receptor (PD-1 DNR), a PD-1 chimeric switch receptor (PD-1 CSR), or a TGF-β dominant negative receptor (TGF-β DNR). [This invention 1044] The isolated nucleic acid molecule of any of claims 1038 to 1043, wherein the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO: 523; and the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO: 524. [This invention 1045] The isolated nucleic acid molecule of any one of claims 1038 to 1044, wherein the TGF-β DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:529. [The present invention 1046] The isolated nucleic acid molecule of any one of claims 1038 to 1045, wherein the polynucleotide sequence encoding the PD-1 DNR, the PD-1 CSR, or the TGF-β DNR is linked to the polynucleotide sequence encoding the CAR via a polynucleotide sequence encoding a 2A self-cleaving peptide. [This invention 1047] The isolated nucleic acid molecule of any one of claims 1038 to 1046, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide. [This invention 1048] The isolated nucleic acid molecule of any of claims 1038 to 1047, comprising, in the 5' to 3' direction, a polynucleotide sequence encoding the CAR, a polynucleotide sequence encoding the 2A self-cleaving peptide, and a polynucleotide sequence encoding the PD-1 DNR, the PD-1 CSR, or the TGF-β DNR. [This invention 1049] Any of the isolated nucleic acid molecules of the present invention 1038 to 1048, which encode a peptide having at least about 95% sequence identity to SEQ ID NO: 521 or 522, or encode a peptide having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 525 to 528. [The present invention 1050] An expression vector comprising any one of the isolated nucleic acid molecules of the present inventions 1038 to 1049. [This invention 1051] A modified immune cell comprising the isolated nucleic acid molecule of any one of 1038 to 1049 of the present invention or the expression vector of 1050 of the present invention. [This invention 1052] The modified immune cell of the present invention 1051, selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and nature killer cells. [This invention 1053] A modified immune cell expressing any one of the CARs of the present invention 1001 to 1023. [This invention 1054] The modified immune cell of the present invention 1053, which also expresses CSR or DNR. [This invention 1055] The modified immune cell of the present invention 1053 or 1054, wherein the CSR is PD-1 CSR and the DNR is PD-1 DNR or TGF-β DNR. [This invention 1056] The modified immune cell of any of claims 1053 to 1055, wherein the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO: 523; and the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO: 524. [This invention 1057] 1057. The modified immune cell of any of claims 1053 to 1056, wherein said TGF-β DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:529. [This invention 1058] The modified immune cell of any of 1053 to 1057, wherein the CAR and the CSR, or the CAR and the DNR, are simultaneously expressed via a 2A self-cleaving peptide. [This invention 1059] The modified immune cell of any of claims 1053 to 1058, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide. [The present invention 1060] The modified immune cell of any of 1053 to 1059 of the present invention, which expresses the CAR and the PD-1 CSR. [The present invention 1061] The modified immune cell of any of claims 1053 to 1060, which expresses the CAR and the TGF-β DNR and is stimulated by a cell expressing DLL3 in the presence of TGF-β. [The present invention 1062] The modified immune cell of any of claims 1053 to 1061 of the present invention, which is selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and nature killer cells. [The present invention 1063] A pharmaceutical composition comprising a CAR of any one of claims 1001 to 1023 of the present invention, a DLL3-binding protein of any one of claims 1024 to 1032 of the present invention or claims 1033 to 1037 of the present invention, an expression vector of claim 1050 of the present invention, or a modified immune cell of any one of claims 1051 to 1062 of the present invention, and a physiologically acceptable excipient. [The present invention 1064] A method for treating a DLL3-related disorder in a subject, comprising administering to the subject a therapeutically effective amount of any of the modified immune cells of present inventions 1051 to 1062, or a therapeutically effective amount of the pharmaceutical composition of present invention 1063. [This invention 1065] 1064. The method of claim 1064, wherein said DLL3-related disorder is a cancer selected from the group consisting of lung cancer, melanoma, breast cancer, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia, and lymphoma. [The present invention 1066] 1065. The method of claim 105, wherein said DLL3-associated disorder is small cell lung cancer. [This invention 1067] The method of any one of claims 1064 to 1066, wherein the cancer expresses DLL3 and PD-L1. [The present invention 1068] The method of any of claims 1064 to 1067, wherein said cancer has a higher TGF-β expression level compared to corresponding normal tissue. [The present invention 1069] Use of a CAR of any of claims 1001 to 1023, a DLL3-binding protein of any of claims 1024 to 1032 or claims 1033 to 1037, an expression vector of claim 1050, or a modified immune cell of any of claims 1051 to 1062, for the preparation of a medicament for treating a DLL3-associated disorder. [The present invention 1070] A drug for use in treating a DLL3-associated disorder, comprising a CAR of any of inventions 1001 to 1023, a DLL3-binding protein of any of inventions 1024 to 1032 or inventions 1033 to 1037, an expression vector of invention 1050, or a modified immune cell of any of inventions 1051 to 1062. [Brief explanation of the drawings]

[0080] [Figure 1] Figure 1 shows a schematic diagram of a VHH-based CAR construct. Sequences that can be used in the construct are listed in SEQ ID NOs: 462-472, 474 and 475. [Figure 2] Figure 2 shows the results of an in vitro cytotoxicity assay of T cells expressing exemplary monospecific CARs containing various camelid anti-DLL3 sdAbs against the small cell lung cancer cell line SHP-77 at an E:T ratio of 2:1 or 5:1. The results for the CARs are presented in order as shown in the legend on the right. [Figure 3]Figure 3 shows the results of cytokine release levels of T cells expressing exemplary monospecific CARs containing various camelid anti-DLL3 sdAbs after co-incubation with the DLL3-expressing tumor cell line SHP-77. IFN-γ and TNF-α release levels (E:T is 2:1 or 5:1) are shown in Figures 3A and 3B, respectively. In each figure, the results for the CARs are presented in order as shown in the legend on the right. [Figure 4] Figure 4 shows the fold expansion of T cells expressing exemplary monospecific CARs comprising various camelid anti-DLL3 sdAbs after long-term stimulation with the small cell lung cancer cell line SHP-77. [Figure 5] Figure 5 shows the in vivo antitumor efficacy results of CAR-T cells expressing a CAR bearing a camel anti-DLL3 sdAb in an SHP-77 tumor model, in which each mouse was injected with a dose of 1 million CAR-T cells. [Figure 6A] Figure 6 shows the results of in vitro cytotoxicity assays of T cells expressing exemplary monospecific CARs containing various humanized camelid anti-DLL3 sdAbs against the small cell lung cancer cell lines SHP-77 (Figure 6A, Figure 6B) and NCI-H82 (Figure 6C, Figure 6D). [Figure 6B] See legend to Figure 6A. [Figure 6C] See legend to Figure 6A. [Figure 6D] See legend to Figure 6A. [Figure 7] Figure 7 shows the results of cytokine release levels of T cells expressing exemplary monospecific CARs containing various humanized camelid anti-DLL3 sdAbs after stimulation with SHP-77. IFN-γ and TNF-α release levels (E:T is 3:1 or 10:1) are shown in Figures 7A and 7B, respectively. In each figure, the results for the CARs are presented in order as shown in the legend on the right. [Figure 8] FIG. 8 shows the fold expansion of T cells expressing exemplary monospecific CARs comprising various humanized camelid anti-DLL3 sdAbs after long-term stimulation with the small cell lung cancer cell line SHP-77. [Figure 9A] Figure 9 shows the in vivo antitumor efficacy results of CAR-T cells expressing a CAR bearing a humanized camelid anti-DLL3 sdAb in an SHP-77 tumor model. In this model, each mouse was injected with a dose of 200,000 CAR-T cells. The results of the nine groups are compared in Figure 9A. The results for each mouse in each group are shown in Figures 9B-9J, respectively. [Figure 9B] See legend to Figure 9A. [Figure 9C] See legend to Figure 9A. [Figure 9D] See legend to Figure 9A. [Figure 9E] See legend to Figure 9A. [Figure 9F] See legend to Figure 9A. [Figure 9G] See legend to Figure 9A. [Figure 9H] See legend to Figure 9A. [Figure 9I] See legend to Figure 9A. [Figure 9J] See legend to Figure 9A. [Figure 10A] Figure 10 shows a schematic diagram of a tandem CAR (Figure 10A) and an armed CAR construct (Figure 10B, Figure 10C). [Figure 10B] See legend to Figure 10A. [Figure 10C] See legend to Figure 10A. [Figure 11A] Figure 11 shows the comparison of the in vitro functional activity of tandem and monospecific CAR-T cells by short-term cytotoxicity (Figures 11A-E, 11V) and cytokine release (Figures 11F-K, 11W-X) and long-term stimulation assays (Figures 11L-U, 11Y-Z). [Figure 11B] See legend to Figure 11A. [Figure 11C] See legend to Figure 11A. [Figure 11D] See legend to Figure 11A. [Figure 11E] See legend to Figure 11A. [Figure 11F] See legend to Figure 11A. [Figure 11G] See legend to Figure 11A. [Figure 11H] See legend to Figure 11A. [Figure 11I] See legend to Figure 11A. [Figure 11J] See legend to Figure 11A. [Figure 11K] See legend to Figure 11A. [Figure 11L] See legend to Figure 11A. [Figure 11M] See legend to Figure 11A. [Figure 11N] See legend to Figure 11A. [Figure 11O] See legend to Figure 11A. [Figure 11P] See legend to Figure 11A. [Figure 11Q] See legend to Figure 11A. [Figure 11R] See legend to Figure 11A. [Figure 11S] See legend to Figure 11A. [Figure 11T] See legend to Figure 11A. [Figure 11U] See legend to Figure 11A. [Figure 11V] See legend to Figure 11A. [Figure 11W] See legend to Figure 11A. [Figure 11X] See legend to Figure 11A. [Figure 11Y] See legend to Figure 11A. [Figure 11Z] See legend to Figure 11A. [Figure 12A] Figure 12 shows the in vitro functional comparison of CAR-T cells and T3s armed with PD-1 DNR or PD-1 CSR targeting SHP-77 cells (Figures 12A and 12B) and SHP-77 / PD-L1 (Figures 12C and 12D), respectively. [Figure 12B] See legend to Figure 12A. [Figure 12C] See legend to Figure 12A. [Figure 12D] See legend to Figure 12A. [Figure 13A] Figure 13 shows that TGF-β-DNR enhances the in vitro and in vivo antitumor efficacy of DLL3 CAR-T cells. Figure 13A shows a schematic diagram of a CAR armed with TGF-β-DNR. Figure 13B shows the positive rates of CAR and TGF-β-DNR for each CAR-T cell. In vitro antitumor efficacy was evaluated in short-term stimulation assays (Figures 13C and 13D) and long-term stimulation assays (Figures 13E-G). Figure 13H shows in vivo antitumor efficacy. Figure 13I shows the pharmacokinetics of CAR-T cells in the peripheral blood of an SHP77 xenograft model after treatment. [Figure 13B] See legend to Figure 13A. [Figure 13C] See legend to Figure 13A. [Figure 13D] See legend to Figure 13A. [Figure 13E] See legend to Figure 13A. [Figure 13F] See legend to Figure 13A. [Figure 13G] See legend to Figure 13A. [Figure 13H] See legend to Figure 13A. [Figure 13I] See legend to Figure 13A. DETAILED DESCRIPTION OF THE INVENTION

[0081] Detailed Description Unless otherwise defined, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art. Any methods, devices, and materials similar or equivalent to those described herein can be used in the practice of the present invention. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not intended to limit the scope of the present disclosure.

[0082] The articles "a" and "an" are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0083] The term "binding protein," as used herein, refers to a molecule or portion of a molecule that binds to a target molecule (e.g., DLL3). In some embodiments, the binding protein comprises an antibody. In some embodiments, the binding protein comprises an antigen-binding fragment of an antibody. In some embodiments, the binding protein may further comprise a low molecular weight component, such as a small molecule drug or toxin. The binding protein may also be an antibody or an antigen-binding fragment thereof. In some embodiments, the binding protein comprises the ligand-binding domain of a receptor. In some embodiments, the binding protein comprises the extracellular domain of a transmembrane receptor. The binding protein may also be the ligand-binding domain of a receptor or the extracellular domain of a transmembrane receptor. In some embodiments, the binding protein comprises a single-domain antibody (sdAb) or a single-chain variable fragment (scFv). In some embodiments, the binding protein may be an sdAb or an scFv. The DLL3-binding protein may be a DLL3-binding domain. In some embodiments, the DLL3-binding protein comprises an antibody or an antigen-binding fragment of an antibody that binds to DLL3. In some embodiments, the DLL3-binding protein may be an antibody or an antigen-binding fragment of an antibody. In some embodiments, the DLL3 binding protein comprises a single domain antibody (sdAb) or a single chain variable fragment (scFv) that binds to DLL3. In some embodiments, the DLL3 binding protein can be an sdAb or an scFv.

[0084] The term "antibody" generally refers to any immunoglobulin (Ig) molecule consisting of four polypeptide chains, two heavy (H) chains and two light (L) chains, or any functional fragment thereof, that retains the essential epitope-binding characteristics of an Ig molecule. In full-length antibodies, each heavy chain consists of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2, and CH3. Each light chain consists of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region consists of one domain, CL. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity-determining regions (CDRs), interspersed with more conserved regions, termed framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The immunoglobulin molecule can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. In a broad sense, the term "antibody" also refers to scFvs or sdAbs that are not derived from immunoglobulin molecules having four polypeptide chains.

[0085] Antibody fragments are portions of antibodies, such as F(ab')2, Fab, Fv, scFv, sdAb, etc. Functional fragments of full-length antibodies retain the target specificity of the full-length antibody. Therefore, recombinant functional antibody fragments, such as scFv (single-chain variable fragment), have been used to develop therapeutics as an alternative to mAb-based therapeutics. scFv fragments (approximately 25 kDa) consist of two variable domains, VH and VL. In nature, the VH and VL domains tend to associate and dissociate non-covalently through hydrophobic interactions. However, stable fragments can be engineered to generate scFvs by linking the domains with a hydrophilic flexible linker.

[0086] As used herein, the term "single domain antibody" (sdAb) has its general meaning in the art and refers to a single heavy chain variable domain of an antibody type that can be found in camelid mammals and that naturally lacks light chains. Such single domain antibodies are characterized by a V H Single domain antibodies are also referred to as "H" or "nanobodies." The amino acid sequence and structure of single domain antibodies can be considered to consist of four framework regions (FR1, FR2, FR3, and FR4) and three complementarity determining regions (CDR1, CDR2, and CDR3). Thus, single domain antibodies can be defined as amino acid sequences with the general structure FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, similar to the variable domains VH or VL. The use of sdAbs as single antigen-binding proteins or as antigen-binding domains in larger proteins or polypeptides offers several significant advantages over the use of conventional antibodies or antibody fragments (e.g., scFvs). The advantages of sdAbs include that only a single domain is required to bind antigen with high affinity and high selectivity; sdAbs are highly stable to denaturing agents or conditions, including heat, pH, and proteases; and sdAbs have access to targets and epitopes inaccessible to conventional antibodies. Typically, sdAbs are produced in camelids, such as llamas, but can also be made synthetically using techniques well known in the art.

[0087] As used herein, the term "humanized sdAb" refers to a humanized sdAb that is a naturally occurring V H It refers to an sdAb in which one or more amino acid residues in the amino acid sequence of the H sequence are replaced by one or more amino acid residues present at the corresponding positions in the VH domain of a normal four-chain antibody of human origin.This can be achieved by methods well known in the art.For example, the FR of the sdAb can be replaced by human variable FR.Therefore, the humanized sdAb has reduced antigenicity when administered to the human body.

[0088] As used herein, the term "heavy chain-only antibody" or "HCAb" refers to a functional antibody that contains a heavy chain but lacks the light chain typically found in four-chain antibodies. Camelids (such as camels, llamas, or alpacas) are known to produce HCAbs.

[0089] "DLL3," also known as "Delta-like ligand 3," is a transmembrane protein involved in the Notch signaling pathway. Initially identified in C. elegans and Drosophila and subsequently shown to be evolutionarily conserved across invertebrates and vertebrates, the Notch signaling pathway is involved in a range of fundamental biological processes, including normal embryonic development, adult tissue homeostasis, and stem cell maintenance. In Drosophila, Notch signaling is primarily mediated by one Notch receptor gene and two ligand genes, known as Serrate and Delta (Wharton et al., 1985; Rebay et al., 1991). In humans, four known Notch receptors and five DSL (Delta-Serrate LAG2) ligands—two homologs of Serrate, known as Jagged 1 and Jagged 2, and three homologs of Delta, termed Delta-like ligands or DLL1, DLL3, and DLL4—are found. In humans, the DLL3 gene is located on chromosome 19q13 and consists of eight exons spanning 9.5 kb. Alternative splicing within the last exon results in two protein isoforms, both of which share 100% overall identity throughout their extracellular and transmembrane domains, differing only in that the longer isoform has an extended cytoplasmic tail.

[0090] As used herein, the terms "specifically bind" or "specifically binding," or any synonym thereof, refer to the ability of a polypeptide, such as a single domain antibody (sdAb), to specifically recognize and detectably bind to a DLL3 molecule as assayed by standard in vitro assays. For example, binding, as used herein, is measured by the ability of an anti-DLL3 polypeptide of the invention to recognize a DLL3 molecule on the cell surface using well-described antigen-antibody binding assays, flow cytometry, and other assays known to those of skill in the art.

[0091] As used herein, the term "expression vector" refers to a nucleic acid construct or sequence produced recombinantly or synthetically using specific nucleic acid elements that enable the transcription and / or expression of another nucleic acid in a host cell. An expression vector can be part of a plasmid, virus, or nucleic acid fragment. In one example, an expression vector is a DNA vector, such as a plasmid, that contains at least one promoter sequence and at least one terminator sequence (e.g., a polyadenylation sequence), optionally an origin of replication (ori) sequence, and optionally a selection or selectable marker sequence. Optionally, the expression vector may further contain at least one nucleotide coding sequence of interest that encodes at least one polypeptide, wherein the at least one promoter sequence is operably linked to the at least one coding sequence. The term "expression" includes any step involved in the production of a polypeptide, including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and / or secretion.

[0092] The term "isolated" refers to material that is substantially or essentially free from components that normally accompany it in its natural state. The material can be a cell or a macromolecule such as a protein or nucleic acid. For example, "isolated nucleic acid," as used herein, refers to a polynucleotide that has been purified from sequences that flank it in its naturally occurring state, e.g., a DNA fragment that has been removed from sequences that normally flank the fragment. Alternatively, "isolated antibody" or "isolated polypeptide," etc., as used herein, refer to the in vitro isolation and / or purification of an antibody or polypeptide molecule from its natural cellular environment and association with other cellular components.

[0093] "Humanized" forms of non-human (e.g., camelid) antibodies are chimeric antibodies which contain minimal sequence derived from non-human immunoglobulin. In some embodiments, humanized antibodies are human immunoglobulins (recipient antibody) in which residues from a hypervariable region (HVR) of the recipient are substituted by residues from an HVR of a non-human species (donor antibody) such as mouse, rat, rabbit, or non-human primate having the desired specificity, affinity, and / or capacity. In some cases, framework ("FR") residues of the human immunoglobulin are substituted by corresponding non-human residues. Furthermore, humanized antibodies may comprise residues which are not found in the recipient antibody or in the donor antibody. These modifications may be made to further refine antibody performance, e.g., binding affinity. Generally, a humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin sequence, and all or substantially all of the FR regions are identical to those of a human immunoglobulin sequence, although the FR regions may include one or more individual FR residue substitutions which improve antibody performance, e.g., binding affinity, isomerization, immunogenicity, etc. The number of these amino acid substitutions in the FRs will typically be no more than six in the H chain and no more than three in the L chain. The humanized antibody will also optionally comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin. For further details, see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). Also see, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409.

[0094] With respect to peptide, polypeptide, or antibody sequences, "sequence identity" and "homology" are defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular peptide or polypeptide sequence after aligning the sequences, introducing gaps as necessary, and achieving the maximum percent sequence identity without considering any conservative substitutions as part of the sequence identity. Alignment, intended to determine percent amino acid sequence identity, can be achieved in a variety of ways within the skill of the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN™ (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

[0095] Dissociation constant (K D or K d ) is used as an index of the affinity of an antibody for an antigen. For example, analysis can be easily performed by the Scatchard method using antibodies labeled with various markers, and by using the OTC measurement kit BiacoreX (manufactured by Amersham Biosciences) or a similar kit in accordance with the user manual and experimental procedures provided with the kit. K that can be derived using these methods D Values ​​are expressed in M ​​(molar) units.

[0096] As used herein, "chimeric switch receptor (CSR)" refers to a receptor engineered to reverse the outcome of an immune cell's (e.g., CAR T cell) original signaling pathway to confer a desired activity on the cell, e.g., the ability to overcome an immunosuppressive tumor microenvironment and to enhance in vivo persistence. In some embodiments, the CSR can utilize inhibitory molecules expressed by cancer cells to further stimulate the CAR T cell. In a non-limiting example, a CAR T cell can be engineered to express a CSR consisting of the extracellular ligand-binding domain of the human inhibitory receptor, programmed cell death protein 1 (PD-1), fused to the transmembrane and cytoplasmic costimulatory signaling domains of CD28. When the CAR T cell is administered to a subject with a cancer that expresses DLL3 and programmed cell death ligand 1 (PD-L1), the expressed CAR can bind to DLL3, and the expressed switch receptor can bind to PD-L1. The properties of the PD-1 / CD28 chimeric switch receptor fusion protein block normal PD1 / PD-L1-mediated T cell suppression and instead promote signaling through the CD28 domain, leading to CAR T cell stimulation. Thus, swapping the transmembrane and intracellular domains of PD-1 with those of CD28 converts PD-L1 into a costimulatory ligand for CAR T cells. This induces enhanced toxicity against cancer cells expressing PD-L1. In other embodiments, CSR can also be used to inhibit the effect of CAR T cells on unintended target cells.

[0097] As used herein, a "dominant negative receptor (DNR)" refers to a receptor that can bind to its ligand but does not induce a signal transduction cascade inside the cell. DNRs typically have an intact ligand-binding domain but lack the intracellular enzymatic domain. They can be a mutant form of the full-length receptor or a truncated form of the receptor. After CAR T cell immunotherapy, some cancers, particularly solid cancers, may upregulate inhibitory ligands that bind to inhibitory receptors on CAR T cells. This adaptive resistance impairs the efficacy of chimeric CAR T cell therapy. Some cancers, particularly solid cancers, are known to secrete transforming growth factor-β (TGF-β), which creates an immunosuppressive environment. TGF-β is known to induce or promote metastasis and potently suppress the immune system. Therefore, in some embodiments, the inventors improve the anti-tumor performance of the CAR T cells disclosed herein by using a truncated version of the TGF-β receptor TGF-βRII as a TGF-β DNR. In some embodiments, the CAR and TGF-β DNR are simultaneously expressed on the surface of T cells by using a 2A self-cleaving peptide. In some embodiments, the CAR and TGF-β DNR are separately expressed on the surface of T cells by using two expression vectors. The inventors have found that when a TGF-β DNR is introduced into the anti-DLL3 CAR T cells disclosed herein, it can enhance the cytotoxicity of the CAR T cells against some DLL3-positive cancer cells, such as SCLC cells. Similarly, in some embodiments, the inventors use a truncated version of the PD-1 receptor as a PD-1 DNR to improve the anti-tumor performance of the CAR T cells disclosed herein.

[0098] As used herein, "treatment" or "treating" includes any beneficial or desired effect on the symptoms or pathology of a disease or condition, and may further include a minimal reduction in one or more measurable markers of the disease or condition being treated, e.g., cancer, autoimmune disease, immune disorder, etc. Treatment may optionally include a delay in the progression of the disease or condition. "Treatment" does not necessarily indicate a complete eradication or cure of the disease or condition or its associated symptoms.

[0099] Some embodiments of the present invention relate to DLL3 binding proteins that have binding specificity for human or rhesus monkey DLL3 protein.

[0100] In some embodiments, the DLL3 binding protein comprises a single domain antibody ("sdAb") portion that specifically binds to DLL3, and the sdAb portion comprises a polypeptide comprising a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81 or a variant thereof comprising up to about three amino acid substitutions in CDR1, a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162 or a variant thereof comprising up to about three amino acid substitutions in CDR2, and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243 or a variant thereof comprising up to about three amino acid substitutions in CDR3.

[0101] In some embodiments, the sdAb comprises a polypeptide comprising a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81, a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162, and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243, or a variant of the polypeptide comprising up to about three amino acid substitutions in the CDR regions. In some embodiments, the anti-DLL3 antibody is or comprises a single domain antibody (sdAb) produced from a camel after immunization with human or rhesus DLL3 protein. In some embodiments, the sdAb comprises a set of CDRs (i.e., a combination of CDR1, CDR2, and CDR3) as listed in each column of Table 1.

[0102] In some embodiments, the sdAb is: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. The present invention also includes a polypeptide comprising any one of the following:

[0103] In some embodiments, the sdAb is: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 The present invention also includes a polypeptide comprising any one of the following:

[0104] In some embodiments, the sdAb comprises an amino acid sequence having at least about 95% (e.g., about 96%, about 97%, about 98%, about 99% or about 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274-354. In some embodiments, the sdAb comprises the amino acid sequence of any one of SEQ ID NOs: 274-354. In other embodiments, the sdAb is humanized and comprises an amino acid sequence having at least about 95% (e.g., about 96%, about 97%, about 98%, about 99% or about 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355-367. In some embodiments, the sdAb comprises the amino acid sequence of any one of SEQ ID NOs: 355-367. Humanized antibodies can be produced using a variety of techniques known in the art, including, but not limited to, CDR grafting (see, e.g., U.S. Pat. Nos. 5,225,539, 5,530,101, and 5,585,089), veneering or resurfacing (see, e.g., European Patent Nos. 592,106 and 519,596), and chain shuffling (see, e.g., U.S. Pat. No. 5,565,332). Generally, during humanization, CDR residues of a recipient antibody (e.g., a human antibody) are replaced with CDR residues from a donor antibody (e.g., a rodent antibody) to retain antigen-binding specificity while minimizing in vivo immunogenicity. Often, framework residues within the framework regions are replaced with the corresponding residues from the donor antibody, resulting in altered, e.g., improved, antigen binding. These framework substitutions, e.g., conservative substitutions, are identified by methods well known in the art, such as modeling the interactions of CDR and framework residues to identify framework residues important for antigen binding and sequence comparison to identify unusual framework residues at particular positions (see, e.g., Queen et al., U.S. Pat. No. 5,585,089; and Riechmann et al., 1988, Nature, 332:323).

[0105] In some cases, an sdAb can be fused to a human IgG hinge fragment and Fc fragment to form a heavy chain antibody (HCAb). In some cases, an sdAb can be fused to another sdAb or scFv specific for an antigen other than DLL3 to form a bispecific antibody. In some cases, an sdAb can be fused to two or more sdAbs or scFvs specific for antigens other than DLL3 to form a multispecific antibody. In other cases, an sdAb can be chemically modified to carry a drug molecule. Thus, an anti-DLL3 sdAb can be used in vivo to target drug molecules to DLL3-expressing cells.

[0106] In some embodiments, the DLL3 binding protein is or comprises a single-chain variable fragment (scFv) that specifically binds to DLL3. In some cases, scFvs are isolated from a synthetic human Fab or scFv phage library through multiple rounds of phage panning, each round of panning including the process of binding, removal of nonspecific phage, and elution and amplification of bound phage in preparation for the next round. In some embodiments, the DLL3 binding protein comprises an scFv portion that specifically binds to DLL3, wherein the scFv comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO:498 or 504, CDR2 set forth in SEQ ID NO:499 or 505, and CDR3 set forth in SEQ ID NO:500 or 506, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO:495 or 501, CDR2 set forth in SEQ ID NO:496 or 502, and CDR3 set forth in SEQ ID NO:497 or 503. In some embodiments, the DLL3 binding protein comprises an scFv portion that specifically binds to DLL3, wherein the scFv comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO:498, CDR2 set forth in SEQ ID NO:499, and CDR3 set forth in SEQ ID NO:500, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO:495, CDR2 set forth in SEQ ID NO:496, and CDR3 set forth in SEQ ID NO:497; or wherein the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO:504, CDR2 set forth in SEQ ID NO:505, and CDR3 set forth in SEQ ID NO:506, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO:501, CDR2 set forth in SEQ ID NO:502, and CDR3 set forth in SEQ ID NO:503.

[0107] In some cases, an scFv can be fused to another sdAb or scFv specific for an antigen other than DLL3 to form a bispecific antibody. In some cases, an scFv can be fused to two or more sdAbs or scFvs specific for antigens other than DLL3 to form a multispecific antibody. In other cases, an scFv can be chemically modified to carry a drug molecule. Thus, an anti-DLL3 scFv can be used in vivo to target drug molecules to DLL3-expressing cells.

[0108] Some embodiments of the present invention relate to a CAR or CAR-T cell that comprises a DLL3 binding domain (an anti-DLL3 CAR or an anti-DLL3 CAR-T cell).

[0109] The CAR of the present invention comprises an extracellular domain, a transmembrane domain, and an intracellular domain. In some embodiments, the CAR further comprises an N-terminal signal peptide and a hinge region between the extracellular domain and the transmembrane domain. The extracellular domain comprises a target-specific binding factor (also referred to as an antigen recognition domain or antigen binding domain). The intracellular domain or otherwise cytoplasmic domain often comprises one or more costimulatory signaling domains and a CD3 zeta chain portion. The costimulatory signaling domain refers to the portion of the CAR that comprises the intracellular domain of a costimulatory molecule.

[0110] Antigen recognition or targeting by a CAR molecule most commonly involves the use of an antibody or antibody fragment. According to the present invention, the antigen-binding domain is an antibody or antibody fragment that specifically binds to DLL3. Preferably, the antigen-binding domain of the CAR of the present invention is an anti-DLL3 scFv or sdAb as described above.

[0111] The transmembrane domain may be derived from either natural or synthetic sources. If the source is natural, the domain may be derived from any membrane-bound or transmembrane protein. A transmembrane region of particular use in the present invention may be derived from (i.e., may comprise at least the transmembrane region of) the alpha, beta, or zeta chain of the T cell receptor, the CD8 α chain, for example.

[0112] The intracellular signaling domain of the CAR of the present invention is involved in activating at least one of the normal effector functions of immune cells. The term "effector function" refers to a specialized function of a cell. For example, the effector function of a T cell can be cytolytic activity or helper activity, including cytokine secretion. Thus, the term "intracellular domain" refers to the portion of a protein that transmits an effector function signal and directs the cell to carry out a specialized function. Typically, the entire cytoplasmic domain can be utilized, although in many cases, it is not necessary to use the entire chain. To the extent that a truncated portion of the cytoplasmic domain is used, such a truncated portion can be used in place of the intact chain, so long as it transmits the effector function signal. Thus, the term intracellular domain is meant to include any truncated portion of the cytoplasmic domain sufficient to transmit the effector function signal. Preferred examples of cytoplasmic domains intended for use in the CAR of the present invention include the cytoplasmic sequences of a T cell receptor (TCR) and a co-receptor that act in conjunction to initiate signal transduction after antigen receptor engagement, as well as any derivatives or variants of these sequences and any synthetic sequences having the same functional capabilities. In some embodiments, the intracellular signaling domain is derived from CD3zeta, FcRgamma, FcRbeta, CD3gamma, CD3delta, CD3epsilon, CD5, CD22, CD79a, CD79b, or CD66d.

[0113] In many cases, the signal generated by the TCR alone is insufficient for full activation of T cells. Therefore, secondary or costimulatory signals are used. Therefore, it can be stated that T cell activation is mediated by two different classes of cytoplasmic signaling sequences: those that initiate antigen-dependent primary activation through the TCR (primary cytoplasmic signaling sequences) and those that act in an antigen-independent manner to provide secondary or costimulatory signals (secondary cytoplasmic signaling sequences). A costimulatory signaling sequence refers to the portion of a CAR that contains the intracellular domain of a costimulatory molecule. A costimulatory molecule is a cell surface molecule other than an antigen receptor or its ligand that is required for an efficient response of lymphocytes to antigens. Examples of such molecules include CD27, CD28, 4-1BB, OX40, CD40, PD-1, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, TNFRSF9, TNFRSF4, TNFRSF8, CD40LG, ITGB2, KLRC2, TNFRSF18, TNFRSF14, HAVCR1, LGALS9, DAP10, DAP12, CD83, ligands for CD83, and combinations thereof.

[0114] The hinge region between the extracellular and transmembrane domains of a CAR generally refers to any oligo- or polypeptide that functions to link the transmembrane domain to the extracellular domain within the polypeptide chain. The hinge region can be up to 300 amino acids, preferably 2-100 amino acids, and most preferably 2-10 amino acids.

[0115] In addition to the antigen-binding domain, transmembrane domain, cytoplasmic domain, and hinge region, the CAR of the present invention may also contain a signal peptide sequence linked to the N-terminus of the CAR. Signal peptide sequences are present at the N-terminus of many secretory proteins and membrane proteins and are typically 15 to 30 amino acids in length. Since many of the above protein molecules have signal peptide sequences, these signal peptides can be used as signal peptides for the CAR of the present invention.

[0116] In some embodiments, the CAR comprises a DLL3 binding domain, which comprises or is derived from a single domain antibody (sdAb) or a single chain variable fragment (scFv).

[0117] In some embodiments, the CAR comprises a DLL3-binding domain, wherein the DLL3-binding domain comprises or is derived from a single domain antibody (sdAb), wherein the sdAb comprises a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81 or a variant thereof comprising up to about three amino acid substitutions in CDR1; a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162 or a variant thereof comprising up to about three amino acid substitutions in CDR2; and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243 or a variant thereof comprising up to about three amino acid substitutions in CDR3. In some embodiments, the CAR comprises a DLL3-binding domain, wherein the DLL3-binding domain comprises or is derived from a single domain antibody (sdAb), wherein the sdAb comprises a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81; a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162; and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243.

[0118] In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain is: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. The present invention also includes an sdAb comprising any one of the following:

[0119] In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain is: (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 The present invention also includes an sdAb comprising any one of the following:

[0120] In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain comprises a camelid sdAb, wherein the sdAb comprises an amino acid sequence having at least about 95% (e.g., 96%, 97%, 98%, 99% or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274-354. In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain comprises a camelid sdAb comprising the amino acid sequence of any one of SEQ ID NOs: 274-354. In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain comprises a humanized sdAb, wherein the sdAb comprises an amino acid sequence having at least about 95% (e.g., 96%, 97%, 98%, 99% or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355-367. In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain comprises a humanized sdAb comprising the amino acid sequence of any one of SEQ ID NOs: 355-367.

[0121] In some embodiments, the CAR comprises a DLL3 binding domain, wherein the DLL3 binding domain comprises or is derived from a single chain variable fragment (scFv), wherein the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO:498, CDR2 set forth in SEQ ID NO:499, and CDR3 set forth in SEQ ID NO:500, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO:495, CDR2 set forth in SEQ ID NO:496, and CDR3 set forth in SEQ ID NO:497; or wherein the VH domain of the scFv comprises CDR1 set forth in SEQ ID NO:504, CDR2 set forth in SEQ ID NO:505, and CDR3 set forth in SEQ ID NO:506, and the VL domain of the scFv comprises CDR1 set forth in SEQ ID NO:501, CDR2 set forth in SEQ ID NO:502, and CDR3 set forth in SEQ ID NO:503. In some embodiments, the VH domain of the scFv comprises the amino acid sequence set forth in SEQ ID NO: 508, and the VL domain of the scFv comprises the amino acid sequence set forth in SEQ ID NO: 507; or the VH domain of the scFv comprises the amino acid sequence set forth in SEQ ID NO: 510, and the VL domain of the scFv comprises the amino acid sequence set forth in SEQ ID NO: 509.

[0122] In some embodiments, a CAR of the invention comprises a camelid sdAb provided herein as a DLL3-binding domain and comprises an amino acid sequence having at least about 95% (e.g., 96%, 97%, 98%, 99% or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 476-484. In some embodiments, a CAR of the invention comprises a camelid sdAb provided herein as a DLL3-binding domain and comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 476-484. In some embodiments, a CAR of the invention comprises a humanized sdAb provided herein as a DLL3-binding domain and comprises an amino acid sequence having at least about 95% (e.g., 96%, 97%, 98%, 99% or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 485-494. In other embodiments, a CAR of the invention comprises a humanized sdAb provided herein as a DLL3-binding domain and comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 485-494.

[0123] In some embodiments, a CAR of the invention comprises a human scFv provided herein as a DLL3-binding domain and comprises an amino acid sequence having at least about 95% (e.g., 96%, 97%, 98%, 99% or 100%) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 515-516. In some embodiments, a CAR of the invention comprises a human scFv provided herein as a DLL3-binding domain and has an amino acid sequence selected from the group consisting of SEQ ID NOs: 515-516.

[0124] In some embodiments, a CAR of the invention comprises, from N- to C-terminus, a signal peptide, a DLL3-binding domain, a hinge region, a transmembrane domain, and a cytoplasmic signaling domain. In a specific embodiment, a CAR of the invention comprises, from N- to C-terminus, a CD8α signal peptide as set forth in SEQ ID NO:465, a DLL3-binding domain, a CD8α hinge domain as set forth in SEQ ID NO:466, a CD8α transmembrane domain as set forth in SEQ ID NO:467, a CD137 cytoplasmic domain as set forth in SEQ ID NO:468, a CD28 cytoplasmic domain as set forth in SEQ ID NO:469, and a CD3ζ cytoplasmic domain as set forth in SEQ ID NO:470.

[0125] Some aspects of the invention relate to isolated nucleic acid molecules encoding an sdAb, scFv, or CAR of the invention. In some embodiments, the nucleic acid molecule encodes a camelid sdAb and comprises a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 368-448. In some embodiments, the nucleic acid molecule encodes a humanized sdAb and comprises a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 449-461. In some embodiments, the nucleic acid molecule encodes the VH and VL domains of an scFv, wherein the sequence encoding the VH domain comprises the polynucleotide sequence of SEQ ID NO: 512 or 514, and the sequence encoding the VL domain comprises the polynucleotide sequence of SEQ ID NO: 511 or 513.

[0126] Some aspects of the present application relate to modified immune cells comprising any one of the CARs provided above, any one of the isolated nucleic acids described above, or any one of the vectors described above. In some embodiments, the modified immune cells are cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and nature killer cells. In some embodiments, the cells comprise an expression vector carrying an isolated nucleic acid molecule of the present invention. Genetically modifying cells with an expression vector to express a polypeptide encoded by a portion of a nucleic acid molecule is a genetic technique well known in the art.

[0127] Some aspects of the present invention relate to the use of the DLL3-binding proteins, anti-DLL3 CARs, nucleic acid molecules, or CAR-T cells of the present invention. In some embodiments, the CAR-T cells of the present invention are formulated as a pharmaceutical composition with a physiologically acceptable excipient. As used herein, "physiologically acceptable excipient" includes, but is not limited to, any adjuvant, carrier, diluent, preservative, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, surfactant, or emulsifier acceptable for use in humans or veterinary medicine. In some embodiments, the CAR-T cells of the present invention or a pharmaceutical composition comprising the same are used to treat a DLL3-associated disorder in a subject. Accordingly, provided are methods for treating a DLL3-associated disorder, comprising administering a therapeutically effective amount of the CAR-T cells or pharmaceutical composition of the present invention to a subject suffering from a DLL3-associated disorder. The "therapeutically effective amount" of an antibody, CAR-T cell, or pharmaceutical composition may vary depending on factors such as the condition, age, sex, and weight of the subject (e.g., patient). The term "therapeutically effective amount" can include an amount that is effective to "treat" a subject. When a therapeutic amount is specified, the exact amount contemplated for administration in a particular embodiment can be determined by a physician in consideration of the subject's condition. In some embodiments, the DLL3-associated disorder is a cancer that expresses DLL3 as a cell surface protein, such as melanoma, breast cancer, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia, and lymphoma. Preferably, the DLL3-associated disorder is lung cancer, particularly small cell lung cancer (SCLC). [Example]

[0128] The examples described herein are not intended to represent all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure.

[0129] Example 1. Animal immunization and antibody library construction In this example, it was demonstrated that immunized camels exhibited good immune responses to human or rhesus DLL3 proteins, and that the acquired immune library exhibited excellent quality.

[0130] animal immunity Immunogens containing the extracellular domain of human DLL3 protein (aa 27-466) with an N-terminal FLAG tag (AdipoGen, AG-40B-0151) or / and DLL3 expression plasmid or DLL3-expressing cells (CHO-K1 / DLL3 or / and CHO-K1 / EGF4) were mixed with adjuvant or PBS and injected into camels. Camels were typically immunized two to four times at weekly to biweekly intervals. After multiple immunizations, the immune response to the target antigen, DLL3, was assessed by serum titration using both enzyme-linked immunosorbent assay (ELISA) and flow cytometry assays.

[0131] Phage display library construction Total RNA was extracted from lymphocytes of immunized camels using TRIZOL® reagent according to the manufacturer's protocol. cDNA was synthesized from the RNA template using the PRIMESCRIPT™ First-Strand cDNA Synthesis Kit with oligo(dT)20 primers according to the manufacturer's protocol. H For the construction of the H phage library, V was isolated from camel cDNA. H H was amplified.

[0132] Example 2. Preparation of anti-DLL3 antibodies The anti-DLL3 antibodies provided herein include single domain antibodies (sdAbs) generated from immune camels or human Fabs isolated from a synthetic human Fab library.

[0133] phage display Phage display libraries were constructed using sdAbs obtained by immunization (immunogens containing the extracellular domain of human DLL3 protein (aa 27-466) with an N-terminal FLAG tag (AdipoGen, AG-40B-0151) or / and DLL3 expression plasmids or DLL3-expressing cells (CHO-K1 / DLL3 or / and CHO-K1 / EGF4)). A separate human Fab phage display library was synthesized. Both phage libraries were rescued and stored after filter sterilization at 4°C for further use. Binding phages were isolated using protein-based and cell-based panning with the two phage libraries. At least one round of panning was performed using both libraries until the percentage of DLL3-specific phage clones reached 30% for both the protein-based and cell-based panning approaches. The output phages from each round were evaluated for the number of total output clones, the percentage of DLL3-positive clones by ELISA, and the sequence diversity of the DLL3-specific clones. Based on these results, the best panning output was selected for high-throughput screening.

[0134] High-throughput screening The selected output phage was used to infect exponentially growing E. coli cells. Double-stranded DNA of the output phage was extracted. For high-throughput screening, the sdAb / Fab insert was excised from the phagemid vector and inserted into an antibody fragment expression vector. The resulting plasmid was used to transform exponentially growing E. coli cells, which were then plated and grown overnight at 37°C. Several thousand colonies were individually picked and grown in 96-deep-well plates containing 1 mL of 2xYT medium. Expression of the antibody fragment was induced by adding 1 mM IPTG.

[0135] The sdAb / Fab proteins in the supernatants were analyzed for their ability to bind to DLL3 ECD protein by ELISA and for their ability to bind to the DLL3-expressing SHP-77 cell line (American Type Culture Collection (ATCC)® CRL-2195™) and CHO-K1 / human DLL3 (in-house) by FACS. All binders were sequenced. Redundant sequences were removed. A total of 81 camelid sdAb and 2 human Fab binders were obtained that bound to both human and rhesus DLL3 protein and cell lines. All these binders have unique amino acid sequences.

[0136] Some of these unique binders were further characterized by surface plasmon resonance (SPR) on a BIAcore T200 instrument (GE Healthcare). The experiment was performed as follows: crude sdAb / Fab proteins were captured on a sensor chip via their affinity tags. A high concentration (100 nM) of human DLL3 was flowed over the sensor chip surface and allowed to bind to the antibody fragments for 300 seconds, after which running buffer was injected to allow the formed complexes to dissociate. The on-rate (ka) and off-rate (kd) were estimated based on a single association and dissociation curve and used to calculate the equilibrium dissociation constant (K D ) was estimated. The binding affinities of some of these unique binders are shown in Table 7.

[0137] The CDR sequences of the anti-DLL3 camelid sdAb are listed in Table 1, and the CDR sequences of the anti-DLL3 human scFv are listed in Table 2.

[0138] Table 1. CDR sequences of anti-DLL3 camel sdAb TIFF2026001061000001.tif118151TIFF2026001061000002.tif242151TIFF2026001061000003.tif243151TIFF2026001061000004.tif208151

[0139] Table 2: CDR sequences of anti-DLL3 human scFv TIFF2026001061000005.tif51151

[0140] The amino acid sequences of the anti-DLL3 sdAbs are listed in Table 3. The CDRs of the sdAbs are underlined. The nucleic acid sequences encoding the anti-DLL3 sdAbs are shown in SEQ ID NOs: 368-448.

[0141] Table 3. Amino acid sequences of anti-DLL3 camel sdAbs TIFF2026001061000006.tif190150TIFF2026001061000007.tif245150TIFF2026001061000008.tif245150TIFF20260010610 00009.tif241150TIFF2026001061000010.tif245150TIFF2026001061000011.tif246150TIFF2026001061000012.tif102150

[0142] The amino acid sequences of the VH and VL domains of anti-DLL3 human scFv are listed in Table 4. The nucleic acid sequences encoding the VH or VL domain of anti-DLL3 human scFv are shown in SEQ ID NOs: 511 to 514.

[0143] Table 4: Amino acid sequence of anti-DLL3 human scFv TIFF2026001061000013.tif95147

[0144] Example 3. Generation of monospecific camel CARs The amino acid sequences of the anti-DLL3 camelid sdAb fragments are presented in Table 3 above, and the nucleic acid sequences of the anti-DLL3 camelid sdAb fragments are presented in SEQ ID NOS: 368-448. CAR constructs (SEQ ID NOS: 476-484) were generated using the sdAb fragments in Table 3 and additional sequences. A reference CAR construct (CAR3) was also generated using the human anti-DLL3 scFv, CAR3 scFv (SEQ ID NOS: 473). The full-length CAR contains, from N- to C-terminus, a CD8α signal peptide (SEQ ID NOS: 465), the DLL3-binding domain sdAb presented in Table 3, a CD8α hinge domain (SEQ ID NOS: 466), a CD8α transmembrane domain (SEQ ID NOS: 467), a CD137 intracellular domain (SEQ ID NOS: 468) or a CD28 intracellular domain (SEQ ID NOS: 469), and a CD3ζ intracellular domain (SEQ ID NOS: 470). A schematic diagram of the CAR construct is shown in Figure 1. The nucleic acid encoding the CAR fragment was then cloned into a lentiviral vector, using the human EF1α promoter for expression to generate a full-length CAR construct in a single coding frame. The resulting CAR backbone vector was designated "PLLV-hEF1α-DLL3."

[0145] Example 4. Generation of camel anti-DLL3 CAR-T cells Preparation of lentivirus A lentiviral packaging plasmid mixture containing pCMV-ΔR-8.47 and pMD2.G (Addgene, catalog no. 12259) was premixed with the PLLV-hEF1α-DLL3 vector at a pre-optimized ratio (1:1:1:2) in the presence of polyethyleneimine before addition to HEK293 cells. After overnight incubation, the supernatant was collected. The virus-containing supernatant was filtered through a 0.45 μm PES filter and ultracentrifuged to concentrate the lentivirus. The viral pellet was rinsed with pre-chilled DPBS. The virus was appropriately aliquoted and immediately stored at -80°C. Viral titers were determined by measuring transduction efficiency in the supT1 cell line via flow cytometry assay.

[0146] T lymphocyte collection and transduction Leukocytes were collected from healthy donors by apheresis. Peripheral blood mononuclear cells (PBMCs) were isolated using Ficoll-Paque™ PLUS medium according to the manufacturer's protocol. Human T cells were purified from PBMCs using a pan T cell isolation kit (Miltenyi, catalog number 130-096-535) according to the manufacturer's protocol. Purified T cells were then preactivated for 48 hours using a human T cell activation / expansion kit (Miltenyi, catalog number 130-091-441) according to the manufacturer's protocol, and anti-CD3 / CD28 MACSiBead particles were added at a bead-to-cell ratio of 1:2. Preactivated T cells were transduced with lentiviral stock in the presence of 7 μg / mL polybrene. The transduced cells were then transferred to a cell culture incubator to allow transgene expression under appropriate conditions.

[0147] Example 5. Evaluation of the in vitro activity of camel anti-DLL3 CAR-T cells In vitro cytotoxicity assay Six days after transduction, transduced T cells were harvested and co-incubated with the DLL3-expressing tumor cell line SHP-77 at effector (CAR-T) to target cell ratios of 2:1 and 5:1 for 20 hours. In all assays, CAR3 CAR-T cells were used as a reference to compare assay variability and / or act as a control. Untransduced T cells (UnT) were used as a negative control.

[0148] The cytotoxicity of the transduced T cells was determined by lactate dehydrogenase (LDH) assay. The results show that CAR3 CAR-T and some anti-DLL3 CAR-T exhibit potent antitumor activity against SHP-77 cells in vitro, while UnT has no targeted cell-killing effect (Figure 2).

[0149] IFN-γ and TNF-α release detection In addition, supernatants from in vitro cytotoxicity assays were collected to assess CAR-induced cytokine release, such as interferon gamma (IFN-γ) and TNF-α release. As shown in Figures 3A and 3B, CAR3 CAR-Ts and some anti-DLL3 CAR-Ts stimulated by SHP-77 produced IFN-γ and TNF-α, whereas UnTs produced little or no IFN-γ or TNF-α. For protocols for detecting IFN-γ and TNF-α release, please refer to CISBIO's human TNF-α kit and IFN-γ kit.

[0150] CAR-T expansion by long-term stimulation assay Day 0, 1 x 10 5 SHP-77 cells were plated in 24-well plates to establish a monolayer. On day 1, transduced T cells were counted and 2 × 10 5 Individual Survival Car + T cells were plated on top of SHP-77 cells in fresh medium in the absence of cytokines. On day 3, fresh 1 x 10 5 A monolayer of SHP-77 cells was plated on top of the CAR-T cells. On day 4, viable CAR-T cells were counted for each well. On the same day, 2 × 10 5 CARs from expanded wells (with at least this amount of cells) + T cells were replated and a new monolayer was established as on day 1. The process was repeated 3–4 times for stimulation. The fold expansion after each stimulation was calculated as [day 4 viable CARs]. + T cells] / 2×10 5 The cumulative expansion was calculated as [(fold expansion) × (fold expansion + 1)] to normalize for cells discarded with each new stimulation.

[0151] After three stimulations, the fold expansion of different CAR-T constructs was calculated. As shown in Figure 4, most CAR-T constructs expanded more than CAR3 CAR-T after three stimulations with SHP-77 tumor cells.

[0152] Example 6. Evaluation of in vivo efficacy of camelid CARs by CAR-T cell-mediated tumor growth inhibition The antitumor activity of camelid CAR was evaluated in the SHP-77 tumor model. SHP-77 cells were implanted subcutaneously into NOD / SCID mice and randomized into seven groups (four mice per group, day 0). Group 1: vehicle (PBS only); Group 2: UnT (negative control); Group 3: CAR3; Group 4: CAS64380; Group 5: CAS64511; Group 6: CAS63931; Group 7: CAS63997. When tumors were palpable (100 mm 3 ), treatment with CAR-T cells, UnT cells, or vehicle (PBS only) was initiated, and their tumor volumes reached approximately 3000 mm 3 Mice were euthanized when tumor volume reached 1 × 10. Tumor volume was measured twice weekly. CAR-T cells were administered at a dose of 1 × 10. 6 CAR-positive T cells / mouse were administered intravenously. After tumor cell implantation, mice and tumors were monitored for approximately 21 days.

[0153] As shown in Figure 5, all selected camelid CARs exhibited antitumor activity in this animal tumor model.

[0154] Example 7. Humanization of camel sdAb Selected camelid sdAbs (SEQ ID NOS: 279, 294, 297, 312) were humanized using CDR-grafting techniques (see, e.g., U.S. Pat. No. 5,225,539). Briefly, camelid sdAb sequences were compared with those available in the Research Collaboratory for Structural Bioinformatics (RCSB) Protein Data Bank. Homology models of each camelid sdAb were generated based on the closest VH structure. From the model structures, residues located near the CDRs or buried in the interior of the molecule (i.e., with less than 15% of the side-chain solvent accessible surface area) were identified.

[0155] Each camelid sdAb sequence was then BLAST searched against the NCBI human germline V gene database to identify the human VH germline sequence (i.e., human acceptor) with the highest identity to the sdAb (see, e.g., Foote and Winter, J. Mol. Biol. 224:487-499 (1992); Morea V. et al., Methods 20:267-279 (2000); Chothia C. et al., J. Mol. Biol. 186:651-663 (1985)). In the CDR-grafting approach, the CDRs of the human acceptor were replaced with those of the camelid sdAb, generating a straight-graft sequence. Straight-grafted antibodies typically lose binding activity, which must be restored by replacing framework residues critical for antibody activity with non-human residues. Amino acid residues located near the CDRs or buried inside the molecule are usually important for the activity and structure of the antibody and therefore needed to be potential backmutation sites. A series of humanized variants were designed using this method. The CDR amino acid sequences of the humanized variants are shown in Table 5. The full-length amino acid sequences of the humanized variants are shown in Table 6. The CDRs are underlined.

[0156] Table 5. CDR sequences of anti-DLL3 humanized sdAbs TIFF2026001061000014.tif124151

[0157] Table 6. Amino acid sequences of anti-DLL3 humanized sdAbs TIFF2026001061000015.tif234147

[0158] The camel and humanized sdAb sequences were fused with human IgG1 hinge and Fc to generate chimeric and humanized HCAb sequences. DNA encoding these HCAbs was synthesized and inserted into the pTT5 vector. HEK293 cells were transfected with the HCAb expression plasmids. Crude HCAb proteins secreted into the culture medium were subjected to SPR affinity measurements as follows: the capture antibody, anti-human Fc pAb (GE Healthcare), was immobilized on a Biacore™ CM5 chip using EDC-activated amine coupling chemistry to approximately 6,000 RU. The HCAb of interest was captured on the sensor chip surface for 300 seconds. Human DLL3 (AdipoGen, AG-40B-0151) was flowed over the sensor chip surface at a series of increasing concentrations. The association and dissociation phases were monitored. Captured antibody and antigen were removed between cycles using 10 mM glycine-HCl, pH 2.0 buffer, confirming a new binding surface for the antigen. The resulting sensorgrams were globally fitted using a 1:1 binding model to determine on- and off-rates (ka and kd, respectively) and affinity (K D ) was calculated.

[0159] The binding affinity of some humanized sdAbs was measured and compared to that of the original camelid sdAb (Table 7). The majority of the humanized antibodies retained the binding affinity of the camelid sdAb. This example demonstrated the success of humanizing sdAbs using our standard protocol. The majority of sdAbs retained their binding affinity after humanization.

[0160] The scFvs had comparable K D values ​​(AS56788 and AS56704 in Table 7).

[0161] Table 7. Monovalent binding affinities of camelid and humanized antibodies and scFvs TIFF2026001061000016.tif126133

[0162] Example 8. Generation of monospecific humanized CARs The amino acid sequences of the anti-DLL3 humanized sdAbs are presented in Table 6 above, and the nucleic acid sequences of the anti-DLL3 humanized sdAbs are listed in SEQ ID NOs: 449-461. Full CAR constructs (SEQ ID NOs: 485-494) were generated using the humanized sdAbs and additional sequences in Table 6. The full-length CARs comprise, from N- to C-terminus, a CD8α signal peptide (SEQ ID NO: 465), the DLL3 binding domain (humanized sdAb) presented in Table 6, a CD8α hinge domain (SEQ ID NO: 466), a CD8α transmembrane domain (SEQ ID NO: 467), a CD137 intracellular domain (SEQ ID NO: 468) or a CD28 intracellular domain (SEQ ID NO: 469), and a CD3ζ cytoplasmic domain (SEQ ID NO: 470). A schematic diagram of the CAR construct is shown in Figure 1. The nucleic acids encoding the CAR fragments were then cloned into a lentiviral vector, generating full-length CAR constructs in a single coding frame using the human EF1α promoter for expression. The resulting CAR backbone vector was named "PLLV-hEF1α-DLL3."

[0163] Example 9. Evaluation of in vitro activity of humanized anti-DLL3 CAR-T cells Similar to the procedures described in Example 4, the efficacy of the humanized CAR was evaluated via CAR-T cell-mediated killing of tumor cells, cytokine release, and long-term stimulation assays.

[0164] In vitro cytotoxicity assay The results are shown in Figures 6A-6D. Our humanized CAR-T demonstrated superior antitumor efficacy in vitro.

[0165] IFN-γ release assay In addition, supernatants from in vitro cytotoxicity assays were collected to assess CAR-induced cytokine release, such as interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α). As shown in Figures 7A and 7B, CAR3 CAR-Ts and some anti-DLL3 CAR-Ts were stimulated by SHP-77 to produce IFN-γ and TNF-α, whereas UnTs produced little to no IFN-γ or TNF-α.

[0166] CART expansion proliferation by long-term stimulation assay Day 0, 1 x 10 5 SHP-77 cells were plated in 24-well plates to establish a monolayer. On day 1, CAR-T cells were counted and 2 × 10 5 Individual Survival Car + T cells were plated on top of SHP-77 cells in fresh medium in the absence of cytokines. On day 3, fresh 1 x 10 5 A monolayer of NCI-H82 cells was plated on top of the CAR-T cells. On day 4, viable CAR-T cells were counted for each well. On the same day, 2 × 10 5 CARs from expanded wells (with at least this amount of cells) + T cells were replated and a new monolayer was established as on day 1. The process was repeated 3–4 times for stimulation. The fold expansion after each stimulation was calculated as [day 4 viable CARs]. + T cells] / 2×10 5 The cumulative expansion was calculated as [(expansion fold) × (expansion fold + 1)] to normalize for cells discarded with each new stimulation.

[0167] After three stimulations, the fold expansion of different CAR-T constructs was calculated. As shown in Figure 8, most CAR-T constructs expanded more than CAR3 CAR-T after three stimulations with SHP-77 tumor cells.

[0168] Example 10. Evaluation of in vivo efficacy of humanized CARs by CAR-T cell-mediated tumor growth inhibition The antitumor activity of humanized CARs was evaluated in the SHP-77 tumor model. SHP-77 cells were implanted subcutaneously into NOD / SCID mice and randomized into 9 groups: vehicle (PBS only), UnT (negative control), CAR3, CAS64380, CAS64380VH5, CAS64511, CAS64511VH5, CAS63997, and CAS63997VH5 (4 mice per group, day 0). When tumors were palpable (100 mm 3), treatment with CAR-T cells, UnT cells, or vehicle (PBS only) was initiated, and their tumor volumes reached approximately 3000 mm 3 Mice were euthanized when tumor volume reached 0.2 × 10. Tumor volume was measured twice weekly. CAR-T cells were administered at 0.2 × 10. 6 CAR-positive T cells / mouse were administered intravenously. After tumor cell implantation, mice and tumors were monitored for approximately 21 days.

[0169] As shown in Figures 9A-9J, compared to the benchmark CAR3, CAS64380VH5 and CAS63997VH5 exhibited superior antitumor activity in this animal model.

[0170] The antitumor activity of these humanized CARs was not found to correlate with their in vitro cell killing efficacy.

[0171] Some of the amino acid and nucleic acid sequences described herein are listed below. Camel sdAb nucleic acid sequence SEQ ID NO: 368 (Camel sdAb AS63930 nucleic acid sequence) TIFF2026001061000017.tif36147 SEQ ID NO: 369 (Camel sdAb AS63932 nucleic acid sequence) TIFF2026001061000018.tif36146 SEQ ID NO: 370 (Camel sdAb AS63951 nucleic acid sequence) TIFF2026001061000019.tif36146 SEQ ID NO: 371 (Camel sdAb AS63984 nucleic acid sequence) TIFF2026001061000020.tif36146 SEQ ID NO: 372 (Camel sdAb AS63987 nucleic acid sequence) TIFF2026001061000021.tif36146 SEQ ID NO: 373 (Camel sdAb AS63997 nucleic acid sequence) TIFF2026001061000022.tif36146 SEQ ID NO: 374 (Camel sdAb AS64047 nucleic acid sequence) TIFF2026001061000023.tif36146 SEQ ID NO: 375 (Camel sdAb AS64052 nucleic acid sequence) TIFF2026001061000024.tif36146 SEQ ID NO: 376 (Camel sdAb AS64062 nucleic acid sequence) TIFF2026001061000025.tif36146 SEQ ID NO: 377 (Camel sdAb AS64072 nucleic acid sequence) TIFF2026001061000026.tif36147 SEQ ID NO: 378 (Camel sdAb AS64097 nucleic acid sequence) TIFF2026001061000027.tif36146 SEQ ID NO: 379 (Camel sdAb AS64114 nucleic acid sequence) TIFF2026001061000028.tif36146 SEQ ID NO: 380 (Camel sdAb AS64123 nucleic acid sequence) TIFF2026001061000029.tif36146 SEQ ID NO: 381 (Camel sdAb AS64130 nucleic acid sequence) TIFF2026001061000030.tif36147 SEQ ID NO: 382 (Camel sdAb AS64137 nucleic acid sequence) TIFF2026001061000031.tif36147 SEQ ID NO: 383 (Camel sdAb AS64142 nucleic acid sequence) TIFF2026001061000032.tif36146 SEQ ID NO: 384 (Camel sdAb AS64154 nucleic acid sequence) TIFF2026001061000033.tif36146 SEQ ID NO: 385 (Camel sdAb AS64160 nucleic acid sequence) TIFF2026001061000034.tif36146 SEQ ID NO: 386 (Camel sdAb AS64228 nucleic acid sequence) TIFF2026001061000035.tif37146 SEQ ID NO: 387 (Camel sdAb AS64300 nucleic acid sequence) TIFF2026001061000036.tif36147 SEQ ID NO: 388 (Camel sdAb AS64380 nucleic acid sequence) TIFF2026001061000037.tif36147 SEQ ID NO: 389 (Camel sdAb AS64395 nucleic acid sequence) TIFF2026001061000038.tif36146 SEQ ID NO: 390 (Camel sdAb AS64443 nucleic acid sequence) TIFF2026001061000039.tif36146 SEQ ID NO: 391 (Camel sdAb AS64511 nucleic acid sequence) TIFF2026001061000040.tif36146 SEQ ID NO: 392 (Camel sdAb AS64536 nucleic acid sequence) TIFF2026001061000041.tif36146 SEQ ID NO: 393 (Camel sdAb AS64597 nucleic acid sequence) TIFF2026001061000042.tif36146 SEQ ID NO: 394 (Camel sdAb AS64617 nucleic acid sequence) TIFF2026001061000043.tif36146 SEQ ID NO: 395 (Camel sdAb AS64634 nucleic acid sequence) TIFF2026001061000044.tif36146 SEQ ID NO: 396 (Camel sdAb AS69498 nucleic acid sequence) TIFF2026001061000045.tif36146 SEQ ID NO: 397 (Camel sdAb AS69500 ​​nucleic acid sequence) TIFF2026001061000046.tif36147 SEQ ID NO: 398 (Camel sdAb AS69527 nucleic acid sequence) TIFF2026001061000047.tif36146 SEQ ID NO: 399 (Camel sdAb AS68280 nucleic acid sequence) TIFF2026001061000048.tif36146 SEQ ID NO: 400 (Camel sdAb AS68355 nucleic acid sequence) TIFF2026001061000049.tif36147 SEQ ID NO: 401 (Camel sdAb AS69443 nucleic acid sequence) TIFF2026001061000050.tif36147 SEQ ID NO: 402 (Camel sdAb AS75376 nucleic acid sequence) TIFF2026001061000051.tif36146 SEQ ID NO: 403 (Camel sdAb AS75387 nucleic acid sequence) TIFF2026001061000052.tif37147 SEQ ID NO: 404 (Camel sdAb AS75695 nucleic acid sequence) TIFF2026001061000053.tif36146 SEQ ID NO: 405 (Camel sdAb AS76169 nucleic acid sequence) TIFF2026001061000054.tif36146 SEQ ID NO: 406 (Camel sdAb AS63931 nucleic acid sequence) TIFF2026001061000055.tif36147 SEQ ID NO: 407 (Camel sdAb AS63937 nucleic acid sequence) TIFF2026001061000056.tif36146 SEQ ID NO: 408 (Camel sdAb AS63948 nucleic acid sequence) TIFF2026001061000057.tif36146 SEQ ID NO: 409 (Camel sdAb AS63956 nucleic acid sequence) TIFF2026001061000058.tif36147 SEQ ID NO: 410 (Camel sdAb AS63965 nucleic acid sequence) TIFF2026001061000059.tif36146 SEQ ID NO: 411 (Camel sdAb AS63993 nucleic acid sequence) TIFF2026001061000060.tif36146 SEQ ID NO: 412 (Camel sdAb AS63999 nucleic acid sequence) TIFF2026001061000061.tif36146 SEQ ID NO: 413 (Camel sdAb AS64006 nucleic acid sequence) TIFF2026001061000062.tif36146 SEQ ID NO: 414 (Camel sdAb AS64057 nucleic acid sequence) TIFF2026001061000063.tif36147 SEQ ID NO: 415 (Camel sdAb AS64060 nucleic acid sequence) TIFF2026001061000064.tif36146 SEQ ID NO: 416 (Camel sdAb AS64071 nucleic acid sequence) TIFF2026001061000065.tif36146 SEQ ID NO: 417 (Camel sdAb AS64093 nucleic acid sequence) TIFF2026001061000066.tif36146 SEQ ID NO: 418 (Camel sdAb AS64118 nucleic acid sequence) TIFF2026001061000067.tif36146 SEQ ID NO: 419 (Camel sdAb AS64120 nucleic acid sequence) TIFF2026001061000068.tif42146 SEQ ID NO: 420 (Camel sdAb AS64124 nucleic acid sequence) TIFF2026001061000069.tif36146 SEQ ID NO: 421 (Camel sdAb AS64135 nucleic acid sequence) TIFF2026001061000070.tif36147 SEQ ID NO: 422 (Camel sdAb AS64163 nucleic acid sequence) TIFF2026001061000071.tif36146 SEQ ID NO: 423 (Camel sdAb AS64182 nucleic acid sequence) TIFF2026001061000072.tif36146 SEQ ID NO: 424 (Camel sdAb AS64183 nucleic acid sequence) TIFF2026001061000073.tif36146 SEQ ID NO: 425 (Camel sdAb AS64207 nucleic acid sequence) TIFF2026001061000074.tif36146 SEQ ID NO: 426 (Camel sdAb AS64276 nucleic acid sequence) TIFF2026001061000075.tif36146 SEQ ID NO: 427 (Camel sdAb AS64336 nucleic acid sequence) TIFF2026001061000076.tif36146 SEQ ID NO: 428 (Camel sdAb AS64346 nucleic acid sequence) TIFF2026001061000077.tif36146 SEQ ID NO: 429 (Camel sdAb AS64420 nucleic acid sequence) TIFF2026001061000078.tif36146 SEQ ID NO: 430 (Camel sdAb AS64473 nucleic acid sequence) TIFF2026001061000079.tif36146 SEQ ID NO: 431 (Camel sdAb AS64475 nucleic acid sequence) TIFF2026001061000080.tif36146 SEQ ID NO: 432 (Camel sdAb AS64513 nucleic acid sequence) TIFF2026001061000081.tif36146 SEQ ID NO: 433 (Camel sdAb AS64562 nucleic acid sequence) TIFF2026001061000082.tif36146 SEQ ID NO: 434 (Camel sdAb AS64583 nucleic acid sequence) TIFF2026001061000083.tif36146 SEQ ID NO: 435 (Camel sdAb AS64594 nucleic acid sequence) TIFF2026001061000084.tif36146 SEQ ID NO: 436 (Camel sdAb AS64605 nucleic acid sequence) TIFF2026001061000085.tif36146 SEQ ID NO: 437 (Camel sdAb AS64606 nucleic acid sequence) TIFF2026001061000086.tif36146 SEQ ID NO: 438 (Camel sdAb AS68121 nucleic acid sequence) TIFF2026001061000087.tif36147 SEQ ID NO: 439 (Camel sdAb AS68170 nucleic acid sequence) TIFF2026001061000088.tif36147 SEQ ID NO: 440 (Camel sdAb AS63964 nucleic acid sequence) TIFF2026001061000089.tif36146 SEQ ID NO: 441 (Camel sdAb AS64116 nucleic acid sequence) TIFF2026001061000090.tif36146 SEQ ID NO: 442 (Camel sdAb AS68270 nucleic acid sequence) TIFF2026001061000091.tif36147 SEQ ID NO: 443 (Camel sdAb AS68320 nucleic acid sequence) TIFF2026001061000092.tif36147 SEQ ID NO: 444 (Camel sdAb AS68351 nucleic acid sequence) TIFF2026001061000093.tif36147 SEQ ID NO: 445 (Camel sdAb AS75378 nucleic acid sequence) TIFF2026001061000094.tif36146 SEQ ID NO: 446 (Camel sdAb AS75383 nucleic acid sequence) TIFF2026001061000095.tif36147 SEQ ID NO: 447 (Camel sdAb AS75751 nucleic acid sequence) TIFF2026001061000096.tif36146 SEQ ID NO: 448 (Camel sdAb AS76422 nucleic acid sequence) TIFF2026001061000097.tif36146

[0172] Humanized camel sdAb nucleic acid sequence SEQ ID NO: 449 (humanized sdAb AS64380VH4 nucleic acid sequence) TIFF2026001061000098.tif36146 SEQ ID NO: 450 (humanized sdAb AS64380VH5 nucleic acid sequence) TIFF2026001061000099.tif36146 SEQ ID NO: 451 (humanized sdAb AS64380VH6 nucleic acid sequence) TIFF2026001061000100.tif36146 SEQ ID NO: 452 (humanized sdAb AS64380VH7 nucleic acid sequence) TIFF2026001061000101.tif36146 SEQ ID NO: 453 (humanized sdAb AS64511VH4 nucleic acid sequence) TIFF2026001061000102.tif36147 SEQ ID NO: 454 (humanized sdAb AS64511VH5 nucleic acid sequence) TIFF2026001061000103.tif36147 SEQ ID NO: 455 (humanized sdAb AS64511VH6 nucleic acid sequence) TIFF2026001061000104.tif36147 SEQ ID NO: 456 (humanized sdAb AS63931VH4 nucleic acid sequence) TIFF2026001061000105.tif36146 SEQ ID NO: 457 (humanized sdAb AS63931VH5 nucleic acid sequence) TIFF2026001061000106.tif36146 SEQ ID NO: 458 (humanized sdAb AS63931VH6 nucleic acid sequence) TIFF2026001061000107.tif36146 SEQ ID NO: 459 (humanized sdAb AS63997VH4 nucleic acid sequence) TIFF2026001061000108.tif36147 SEQ ID NO: 460 (humanized sdAb AS63997VH5 nucleic acid sequence) TIFF2026001061000109.tif36147 SEQ ID NO: 461 (humanized sdAb AS63997VH6 nucleic acid sequence) TIFF2026001061000110.tif36147 SEQ ID NO: 462 (linker amino acid sequence) TIFF2026001061000111.tif3128 SEQ ID NO: 463 (linker amino acid sequence) TIFF2026001061000112.tif3128 SEQ ID NO: 464 (linker amino acid sequence) TIFF2026001061000113.tif3128 SEQ ID NO: 465 (CD8α signal peptide amino acid sequence) TIFF2026001061000114.tif3128 SEQ ID NO: 466 (CD8α hinge amino acid sequence) TIFF2026001061000115.tif9146 SEQ ID NO: 467 (CD8α transmembrane domain amino acid sequence) TIFF2026001061000116.tif3128 SEQ ID NO: 468 (4-1BB intracellular domain amino acid sequence) TIFF2026001061000117.tif4128 SEQ ID NO: 469 (CD28 intracellular domain amino acid sequence) TIFF2026001061000118.tif4128 SEQ ID NO: 470 (CD3ζ intracellular domain amino acid sequence) TIFF2026001061000119.tif9147 SEQ ID NO: 471 (F2A element amino acid sequence) TIFF2026001061000120.tif4128 SEQ ID NO: 472 (P2A element amino acid sequence) TIFF2026001061000121.tif4128 SEQ ID NO: 473 (CAR3 anti-DLL3 scFv amino acid sequence) TIFF2026001061000122.tif26147 SEQ ID NO: 474 (CD28 transmembrane domain amino acid sequence) TIFF2026001061000123.tif3128 SEQ ID NO: 475 (CD28 hinge) TIFF2026001061000124.tif3128

[0173] Camel anti-DLL3 CAR sequence SEQ ID NO: 476 (CAS63997) TIFF2026001061000125.tif37145 Sequence number 477 (CAS64380) TIFF2026001061000126.tif37146 Sequence number 478 (CAS64511) TIFF2026001061000127.tif37147 sequence number 479 (CAS64617) TIFF2026001061000128.tif35147 sequence number 480 (CAS69443) TIFF2026001061000129.tif37147 Sequence number 481 (CAS63931) TIFF2026001061000130.tif37145 sequence number 482 (CAS64047) TIFF2026001061000131.tif37147 sequence number 483 (CAS64052) TIFF2026001061000132.tif37146 Sequence number 484 (CAS64062) TIFF2026001061000133.tif37147

[0174] Humanized anti-DLL3 CAR sequence SEQ ID NO: 485 (CAS64380VH4) TIFF2026001061000134.tif37145 Sequence number 486 (CAS64380VH5) TIFF2026001061000135.tif37145 Sequence number 487 (CAS64380VH6) TIFF2026001061000136.tif37146 Sequence number 488 (CAS64380VH7) TIFF2026001061000137.tif37146 Sequence number 489 (CAS64511VH4) TIFF2026001061000138.tif37146 SEQ ID NO: 490 (CAS64511VH5) TIFF2026001061000139.tif37147 Sequence number 491 (CAS64511VH6) TIFF2026001061000140.tif37146 SEQ ID NO: 492 (CAS63997VH4) TIFF2026001061000141.tif37146 Sequence number 493 (CAS63997VH5) TIFF2026001061000142.tif37146 SEQ ID NO: 494 (CAS63997VH6) TIFF2026001061000143.tif37146

[0175] Anti-DLL3 human scFv VL and VH domain nucleic acid sequences SEQ ID NO: 511 (nucleic acid sequence of the VL domain of anti-DLL3 human scFv AS56704) TIFF2026001061000144.tif31146 SEQ ID NO: 512 (nucleic acid sequence of the VH domain of anti-DLL3 human scFv AS56704) TIFF2026001061000145.tif36146 SEQ ID NO: 513 (nucleic acid sequence of the VL domain of anti-DLL3 human scFv AS56788) TIFF2026001061000146.tif31146 SEQ ID NO: 514 (nucleic acid sequence of the VH domain of anti-DLL3 human scFv AS56788) TIFF2026001061000147.tif36146

[0176] Human anti-DLL3 scFv CAR sequence SEQ ID NO: 515 (CAS56704) TIFF2026001061000148.tif48147 sequence number 516 (CAS56788) TIFF2026001061000149.tif48147

[0177] Anti-DLL3 Benchmark CAR SEQ ID NO: 517 (1H2.1 amino acid sequence) TIFF2026001061000150.tif42146

[0178] Example 11. Evaluation of the in vitro activity of humanized anti-DLL3 tandem CAR-T cells To improve the anti-tumor efficacy of CAR-T, we constructed three tandem CARs (T1, T2, and T3). The amino acid sequences of the tandem CARs are shown in SEQ ID NOs: 518-520. The amino acid sequences of anti-DLL3 humanized sdAb fragments are shown in SEQ ID NOs: 356 (AS64380VH5) and 366 (AS63997VH5). A reference CAR construct was also generated using the anti-DLL3 CAR, 1H2.1 (SEQ ID NO: 517; see, e.g., WO 2019200007). The full-length CAR comprises, from N- to C-terminus, a CD8α signal peptide (SEQ ID NO: 465), a DLL3-binding domain sdAb represented by SEQ ID NO: 356 (AS64380VH5) and SEQ ID NO: 366 (AS63997VH5), a CD8α hinge domain (SEQ ID NO: 466), a CD8α transmembrane domain (SEQ ID NO: 467), a CD137 intracellular domain (SEQ ID NO: 468) or a CD28 intracellular domain (SEQ ID NO: 469), and a CD3ζ intracellular domain (SEQ ID NO: 470). A schematic diagram of the CAR construct is shown in Figure 10A. For T1, both sdAb1 and sdAb2 were AS64380VH5. For T2, both sdAb1 and sdAb2 were AS63997VH5. For T3, sdAb1 and sdAb2 were AS63997VH5 and AS64380VH5, respectively. The nucleic acid encoding the CAR fragment was then cloned into a lentiviral vector, using the human EF1α promoter for expression to generate a full-length CAR construct in a single coding frame.

[0179] SEQ ID NO: 518 (T1 amino acid sequence) TIFF2026001061000151.tif48147 SEQ ID NO: 519 (T2 amino acid sequence) TIFF2026001061000152.tif48146 SEQ ID NO: 520 (T3 amino acid sequence) TIFF2026001061000153.tif48146

[0180] In vitro cytotoxicity assay Nine days after transduction, transduced T cells were harvested and co-incubated with DLL3-expressing tumor cell lines (SHP-77, with high DLL3 expression, NCI-H82, with moderate DLL3 expression, and NCI-H2171, with low DLL3 expression) and DLL3-negative expressing cell lines (NCI-H460 and HEK293) at effector (CAR-T) to target cell ratios of 0.5:1 and 2:1 for 22 hours. In all assays, CAR3 CAR-T cells were used as a reference to compare assay variability and / or act as a control. Untransduced T cells (UnT) were used as a negative control.

[0181] The cytotoxicity of the transduced T cells was determined by lactate dehydrogenase (LDH) assay. The results show that CAR3 CAR-T and some anti-DLL3 tandem CAR-T exhibited potent antitumor activity against SHP-77 cells in vitro, while UnT had no targeted cell-killing effect (Figures 11A-C), and DLL3-negative expressing cells (NCI-H460 and HEK293) did not induce cytotoxicity (Figures 11D-E). In addition to CAR3, we also compared the in vitro cytotoxicity of T3 and 1H2.1 against SHP-77 cells. The results show that T3 had comparable or less potent cell-killing activity in short-term stimulation (Figure 11V).

[0182] IFN-γ and TNF-α release detection In addition, supernatants from in vitro cytotoxicity assays were collected to assess CAR-induced cytokine release, such as interferon gamma (IFN-γ) and TNF-α release. As shown in Figure 11F-K, CAR3 CAR-Ts and some anti-DLL3 tandem CAR-Ts produced IFN-γ and TNF-α when stimulated by DLL3-expressing cell lines, whereas UnTs produced little or no IFN-γ or TNF-α. DLL3-negative expressing cell lines did not reduce the specific release of IFN-γ and TNF-α (data not shown). For protocols for detecting IFN-γ and TNF-α release, please refer to CISBIO's Human TNF-α Kit and IFN-γ Kit.

[0183] Compared with 1H2.1, T3 released more IFN-γ and TNF-α (after 22 h of co-incubation) (FIGS. 11W-X).

[0184] Tandem CAR-T cytotoxicity and expansion by long-term stimulation assay We evaluated DLL3 CAR-T cells by repeated antigen stimulation assays. Upon repeated stimulation with SCLC and control cell lines, tandem CAR-T cells T3 exhibited more potent cytotoxicity against SCLC cells, particularly SHP-77 and NCI-H82 cells (Figures 11L-P). In addition to cytotoxic activity, tandem CAR-T cells T3 also exhibited higher proliferation potential than other CAR-T cells, particularly upon stimulation with SHP-77 and NCI-H82 cells (Figures 11Q-U).

[0185] In addition to CAR3, we also compared the in vitro cytotoxicity of T3 and 1H2.1 against SHP-77 cells. The results show that T3 had superior cytotoxicity and expanded proliferation in long-term stimulation (Figures 11Y-Z).

[0186] Repeated stimulation was performed as follows. Round 1: CAR-T cells and 3 × 10 5 Target cells (e.g., SHP-77) were added to a 24-well plate at a 1:5 effector-to-target cell ratio and co-incubated for 3 days in a carbon dioxide incubator at 37°C and 5% CO. For cytokine detection, 200 μL of cell culture supernatant was pipetted to collect the co-incubated cells, and the %CD3 and CAR positivity rates were evaluated by flow cytometry. Round 2: Based on the CAR-T positivity rate of the collected cells in Round 1, the collected cells were continued to be co-incubated with the same volume of fresh target cells (SHP-77) at an effector-to-target cell ratio of 1:2 for another 3 days. For cytokine detection, 200 μL of cell culture supernatant was pipetted, and the co-incubated cells were collected, and the %CD3 and CAR positivity rates were evaluated by flow cytometry; Round 3 and subsequent rounds were conducted in a similar manner to Round 2, based on the CAR-T positivity rate of each preceding round.

[0187] Example 12. Evaluation of the in vitro activity of humanized anti-DLL3 CAR-T cells armed with PD-1 DNR or CSR To improve the durability of CAR-T, we constructed DLL3 CARs armed with a PD-1 dominant-negative receptor (PD-1DNR) or a PD-1 chimeric switch receptor (PD-1CSR). The amino acid sequences of the two CARs are presented in SEQ ID NOs: 521-522. The PD-1DNR and PD-1CSR sequences were linked to the C-terminus of T3 via P2A. The amino acid sequences of PD-1DNR and PD-1CSR are presented in SEQ ID NOs: 523-524. A schematic diagram of the CAR construct is shown in Figure 10B.

[0188] SEQ ID NO: 521 (T3-PD-1DNR amino acid sequence) TIFF2026001061000154.tif75146 SEQ ID NO: 522 (T3-PD-1CSR amino acid sequence) TIFF2026001061000155.tif70146 SEQ ID NO: 523 (PD-1DNR amino acid sequence) TIFF2026001061000156.tif26146 SEQ ID NO: 524 (PD-1CSR amino acid sequence) TIFF2026001061000157.tif20146

[0189] We evaluated DLL3 CAR-T cells armed with PD-1DNR or PD-1CSR in a repeated antigen stimulation assay. When repeatedly stimulated with SHP-77 cells, the armed CAR-T cells did not exhibit enhanced cytotoxicity or improved expansion capacity compared with standard CAR-T cells (Figures 12A-B). When repeatedly stimulated with SHP-77 / PD-L1 cells (overexpressing human PD-L1 in SHP-77 cells), CAR-T cells armed with PD-1CSR exhibited superior cytotoxicity and expansion capacity (Figures 12C-D).

[0190] Example 13. TGF-β-DNR enhances the antitumor efficacy of DLL3 CAR-T cells Construction of TGF-β-DNR-armed DLL3 CAR-T cells To improve the antitumor performance of DLL3 CAR-T cells within the tumor microenvironment, we incorporated a TGF-β-DNR sequence into the DLL3 CAR, as shown in Figure 13A. TGF-β-DNR is a truncated version of TGFBRII, consisting of the extracellular and transmembrane domains of TGFBRII. Constructs T3-P2A-TGF-β-DNR and T3-T2A-TGF-β-DNR contain a T3-BBZ sequence at the N-terminus, a P2A or T2A peptide as shown, and a TGF-β-DNR sequence at the C-terminus. Constructs TGF-β-DNR-P2A-T3 and TGF-β-DNR-T2A-T3 contain a TGF-β-DNR sequence at the N-terminus, a P2A or T2A peptide as shown, and a T3-BBZ sequence at the C-terminus. The detailed sequences of the TGF-β-DNR and armed DLL3 CAR are provided in SEQ ID NOs: 525-529.

[0191] SEQ ID NO: 525 (T3-P2A-TGF-β-DNR amino acid sequence) TIFF2026001061000158.tif70146 SEQ ID NO: 526 (TGF-β-DNR-P2A-T3 amino acid sequence) TIFF2026001061000159.tif70147 SEQ ID NO: 527 (T3-T2A-TGF-β-DNR amino acid sequence) TIFF2026001061000160.tif70147 SEQ ID NO: 528 (TGF-β-DNR-T2A-T3 amino acid sequence) TIFF2026001061000161.tif70147 SEQ ID NO: 529 (TGF-β-DNR amino acid sequence) TIFF2026001061000162.tif20146

[0192] All constructs were packaged into lentiviruses based on a second-generation lentivirus system. Primary T cells isolated from PBMCs of healthy donors were then transduced with the lentiviruses. Four days after transduction, the sdAb and TGF-β-DNR positivity rates were detected by FACS (Figure 13B). The results showed that the sdAb positivity rates of T3-P2A-TGF-β-DNR, T3-T2A-TGF-β-DNR, TGF-β-DNR-P2A-T3, and TGF-β-DNR-T2A-T3 CAR-T cells were comparable. However, the positivity rates of TGF-β-DNR were higher for the constructs T3-P2A-TGF-β-DNR and T3-T2A-TGF-β-DNR than for TGF-β-DNR-P2A-T3 and TGF-β-DNR-T2A-T3. These results indicated that the expression of TGF-β-DNR was higher when conjugated to the C-terminus of CAR.

[0193] In vitro cytotoxicity assay The cytotoxicity of these CAR-T cells was then assessed by LDH or IFN-γ release assays. Five days after transduction, CAR-T cells were regulated by untransduced T cells (UnT) to the same sdAb positivity rate. Next, CAR-T cells or UnT cells were co-incubated with SHP77 cells in the presence of 5 ng / mL TGF-β for 48 hours, and LDH and IFN-γ release were measured (Figure 13C-D). The results showed that CAR-T cells armed with TGF-β-DNR induced more specific lysis of target cells than unarmed CAR-T cells. Therefore, CAR-T cells armed with TGF-β-DNR showed a higher IFN-γ release capacity upon antigen activation. Consistent with the TGF-β-DNR expression levels of different CAR-T cells, T3-P2A-TGF-β-DNR and T3-T2A-TGF-β-DNR secreted higher levels of IFN-γ than TGF-β-DNR-P2A-T3 and TGF-β-DNR-T2A-T3 CAR-T cells. Collectively, these results demonstrated that TGF-β-DNR can enhance the cytotoxicity of DLL3 CAR-T cells against DLL3-positive SCLC cells.

[0194] Long-term stimulation assay To determine whether TGF-β-DNR could counteract the inhibitory effects of TGF-β on CAR-T cells, we performed a long-term stimulation assay. Specifically, T3-P2A-TGF-β-DNR and T3 CAR-T cells were repeatedly loaded with SHP77 cells every 3 days in the presence or absence of 5 ng / mL TGF-β. At the end of each stimulation, the percentage of T cells among total viable cells was analyzed by FACS, and CAR-T cell expansion was calculated. As shown in Figures 13E and 13F, the persistence and expansion of T3 CAR-T cells was inhibited by TGF-β. In contrast, the persistence and expansion of T3-P2A-TGF-β-DNR were fully maintained even in the presence of TGF-β. After two rounds of stimulation with SHP77 cells, T cell exhaustion markers were analyzed by FACS. As shown in Figure 13G, treatment with TGF-β upregulated the expression of exhaustion markers in T3 cells but not in T3-P2A-TGF-β-DNR CAR-T cells. Collectively, these results demonstrated that TGF-β-DNR protects DLL3 CAR-T cells from TGF-β inhibition. Because TGF-β expression levels are typically elevated in the microenvironment of solid tumors, our results suggest that the addition of TGF-β-DNR improves the antitumor efficacy of DLL3 CAR-T cells in solid tumors.

[0195] In vivo antitumor efficacy studies To further explore whether TGF-β-DNR could enhance the antitumor efficacy of DLL3 CAR-T cells in vivo, T3-T2A-TGF-β-DNR CAR-T cells or parental CAR-T cells were evaluated in a xenograft model. 7 SHP77 cells were subcutaneously implanted into NCG mice. After 7–10 days, tumor volumes reached 100–200 mm. 3 When it reaches 2.5 × 10 5CAR-T cells were intravenously injected into mice. Tumor volume was then measured twice weekly, and the percentage of CAR-T in peripheral blood was measured weekly. As shown in Figure 13H, at suboptimal doses, T3-T2A-TGF-β-DNR CAR-T cells were able to potently suppress tumor growth, whereas parental T3 CAR-T cells were unable to do so. As shown in Figure 13I, the percentage of T3-T2A-TGF-β-DNR CAR-T in peripheral blood leukocytes was higher than that of T3. Collectively, these results indicate that TGF-β-DNR could enhance the antitumor efficacy of DLL3 CAR-T cells in vivo.

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85 Thr Ile Val Ser Gly Gly Gly Thr Thr Tyr Tyr Ala Asp Ser Val Arg 1 5 10 15 Gly <210> 86 <211> 17 <212> PRT <213> Artificial <220> <223> AS63987 CDR2 <400> 86 Val Ile Tyr Thr Pro Ser Gly Ile Thr Asp Tyr Ala Ser Ser Val Lys 1 5 10 15 Gly <210> 87 <211> 17 <212> PRT <213> Artificial <220> <223> AS63997 CDR2 <400> 87 Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 88 <211> 17 <212> PRT <213> Artificial <220> <223> AS64047 CDR2 <400> 88 Ala Val Tyr Thr Gly Asp Gly Ile Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 89 <211> 17 <212> PRT <213> Artificial <220> <223> AS64052 CDR2 <400> 89 Thr Ile His Ser Gly Val Ala Thr Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 90 <211> 17 <212> PRT <213> Artificial <220> <223> AS64062 CDR2 <400> 90 Ala Leu Tyr Thr Gly Gly Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 91 <211> 17 <212> PRT <213> Artificial <220> <223> AS64072 CDR2 <400> 91 Ala His Gly Pro Val Ser Gly Thr Ala Tyr Tyr Thr Asp Ser Val Lys 1 5 10 15 Gly <210> 92 <211> 17 <212> PRT <213> Artificial <220> <223> AS64097 CDR2 <400> 92 Ala Ile Asn Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Leu Glu 1 5 10 15 Gly <210> 93 <211> 17 <212> PRT <213> Artificial <220> <223> AS64114 CDR2 <400> 93 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val Arg 1 5 10 15 Gly <210> 94 <211> 17 <212> PRT <213> Artificial <220> <223> AS64123 CDR2 <400> 94 Ile Ile Tyr Thr Gly Ser Gly Ser Thr His Tyr Arg Ser Ser Val Lys 1 5 10 15 Gly <210> 95 <211> 17 <212> PRT <213> Artificial <220> <223> AS64130 CDR2 <400> 95 Thr Ile Asp Ser Arg Gly Thr Ile Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 96 <211> 17 <212> PRT <213> Artificial <220> <223> AS64137 CDR2 <400> 96 Thr Leu Val Thr Trp Val Glu Arg Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 97 <211> 17 <212> PRT <213> Artificial <220> <223> AS64142 CDR2 <400> 97 Cys Ile Asp Trp Thr Gly Ala Asn Ile Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 98 <211> 17 <212> PRT <213> Artificial <220> <223> AS64154 CDR2 <400> 98 Cys Ile Tyr Thr Gly Ser Gly Ser Thr Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 99 <211> 17 <212> PRT <213> Artificial <220> <223> AS64160 CDR2 <400> 99 Ala Met Cys Phe Gly Gly Leu Val Thr His Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 100 <211> 16 <212> PRT <213> Artificial <220> <223> AS64228 CDR2 <400> 100 Arg Ile Gln Pro Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 101 <211> 17 <212> PRT <213> Artificial <220> <223> AS64300 CDR2 <400> 101 Tyr Ile Asn Thr Ile Ile Gly Asn Thr Tyr Tyr Thr Asp Ser Val Lys 1 5 10 15 Gly <210> 102 <211> 17 <212> PRT <213> Artificial <220> <223> AS64380 CDR2 <400> 102 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val Arg 1 5 10 15 Gly <210> 103 <211> 17 <212> PRT <213> Artificial <220> <223> AS64395 CDR2 <400> 103 Thr Leu Val Thr Trp Ala Glu Arg Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 104 <211> 17 <212> PRT <213> Artificial <220> <223> AS64443 CDR2 <400> 104 Ile Ile Tyr Thr Gly Gly Glu Ser Thr His Tyr Arg Ser Ser Val Lys 1 5 10 15 Gly <210> 105 <211> 17 <212> PRT <213> Artificial <220> <223> AS64511 CDR2 <400> 105 Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 106 <211> 17 <212> PRT <213> Artificial <220> <223> AS64536 CDR2 <400> 106 Leu Ile Tyr Pro Gly Gly Gly Ser Thr Tyr Tyr Ala Ser Ser Val Lys 1 5 10 15 Gly <210> 107 <211> 17 <212> PRT <213> Artificial <220> <223> AS64597 CDR2 <400> 107 Thr Ile Asp Ser Gly Val Gly Thr Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 108 <211> 16 <212> PRT <213> Artificial <220> <223> AS64617 CDR2 <400> 108 Arg Ile Ser Thr Ser Gly Phe Thr Asn Tyr Ala Ala Ser Val Lys Gly 1 5 10 15 <210> 109 <211> 17 <212> PRT <213> Artificial <220> <223> AS64634 CDR2 <400> 109 Val Phe Tyr Pro Gly Gly Gly Ser Thr Asn Tyr Ala Asp Ser Ala Lys 1 5 10 15 Gly <210> 110 <211> 17 <212> PRT <213> Artificial <220> <223> AS69498 CDR2 <400> 110 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Ile Asp Ser Val Arg 1 5 10 15 Gly <210> 111 <211> 18 <212> PRT <213> Artificial <220> <223> AS69500 CDR2 <400> 111 Ser Ile Phe Thr Gly Thr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 1 5 10 15 Lys Gly <210> 112 <211> 17 <212> PRT <213> Artificial <220> <223> AS69527 CDR2 <400> 112 Ala Ile Tyr Thr Asp Asp Gly Ser Thr Trp Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 113 <211> 17 <212> PRT <213> Artificial <220> <223> AS68280 CDR2 <400> 113 Ala Ile Ser Thr Gly Asp Gly Ala Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 114 <211> 16 <212> PRT <213> Artificial <220> <223> AS68355 CDR2 <400> 114 Ala Ile Asp Ser Asp Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 115 <211> 16 <212> PRT <213> Artificial <220> <223> AS69443 CDR2 <400> 115 Thr Ile Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 116 <211> 17 <212> PRT <213> Artificial <220> <223> AS75376 CDR2 <400> 116 Val Ile Tyr Thr Gly Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 117 <211> 17 <212> PRT <213> Artificial <220> <223> AS75387 CDR2 <400> 117 Val Ala Tyr Thr Gly Gly Asp Ile Gln Tyr Leu Thr Asp Ser Val Lys 1 5 10 15 Gly <210> 118 <211> 16 <212> PRT <213> Artificial <220> <223> AS75695 CDR2 <400> 118 Phe Ile Asp Ala Gly Gly Ala Thr Ile Tyr Ala Asp Pro Val Lys Gly 1 5 10 15 <210> 119 <211> 16 <212> PRT <213> Artificial <220> <223> AS76169 CDR2 <400> 119 Tyr Ile Arg Asp Asn Ile Met Thr Ser Tyr Thr Asp Ser Val Lys Gly 1 5 10 15 <210> 120 <211> 17 <212> PRT <213> Artificial <220> <223> AS63931 CDR2 <400> 120 Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val Thr 1 5 10 15 Gly <210> 121 <211> 17 <212> PRT <213> Artificial <220> <223> AS63937 CDR2 <400> 121 Cys Ile His Thr Gly Leu Gly Arg Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 122 <211> 17 <212> PRT <213> Artificial <220> <223> AS63948 CDR2 <400> 122 Cys Ile Tyr Thr Gly Ser Gly Ser Thr Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 123 <211> 16 <212> PRT <213> Artificial <220> <223> AS63956 CDR2 <400> 123 Leu Ile Asn Ser Ser Gly Gly Thr Tyr Tyr Ala Asp Ser Val Arg Gly 1 5 10 15 <210> 124 <211> 16 <212> PRT <213> Artificial <220> <223> AS63965 CDR2 <400> 124 Lys Val Ile Ser Asp Gly Thr Thr Val Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 125 <211> 16 <212> PRT <213> Artificial <220> <223> AS63993 CDR2 <400> 125 Leu Val Ala Thr Ala Gly Asn Ser Val Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 126 <211> 17 <212> PRT <213> Artificial <220> <223> AS63999 CDR2 <400> 126 Ile Ile Tyr Thr Gly Gly Ile Ser Thr His Tyr Arg Ser Ser Val Lys 1 5 10 15 Gly <210> 127 <211> 17 <212> PRT <213> Artificial <220> <223> AS64006 CDR2 <400> 127 Leu Ile Tyr Thr Ser Gly Thr Ser Glu Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 128 <211> 17 <212> PRT <213> Artificial <220> <223> AS64057 CDR2 <400> 128 Cys Ile Ser Trp Thr Gly Ala Asn Ile Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 129 <211> 16 <212> PRT <213> Artificial <220> <223> AS64060 CDR2 <400> 129 Val Ile Asp Ser Gly Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 130 <211> 17 <212> PRT <213> Artificial <220> <223> AS64071 CDR2 <400> 130 Phe Ile Val Met Ile Arg Gly Thr Thr Tyr Tyr Gly Ala Ser Val Lys 1 5 10 15 Gly <210> 131 <211> 17 <212> PRT <213> Artificial <220> <223> AS64093 CDR2 <400> 131 Ser Ile Tyr Pro Ala Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 132 <211> 16 <212> PRT <213> Artificial <220> <223> AS64118 CDR2 <400> 132 Phe Ile Asn Ser Ala Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 133 <211> 17 <212> PRT <213> Artificial <220> <223> AS64120 CDR2 <400> 133 Ser Ile Tyr Pro Ala Gly Gly Arg Pro Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 134 <211> 17 <212> PRT <213> Artificial <220> <223> AS64124 CDR2 <400> 134 Ala Ile Ala Thr Ile Gly Gly Thr Thr Arg Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 135 <211> 16 <212> PRT <213> Artificial <220> <223> AS64135 CDR2 <400> 135 Ala Met Gln Pro Asp Gly Thr Thr Ser Tyr Ser Asp Ser Val Lys Gly 1 5 10 15 <210> 136 <211> 17 <212> PRT <213> Artificial <220> <223> AS64163 CDR2 <400> 136 Ala Ile Ser Thr Gly Ser Gly Ser Thr Tyr Tyr Ala Gly Ser Val Lys 1 5 10 15 Gly <210> 137 <211> 17 <212> PRT <213> Artificial <220> <223> AS64182 CDR2 <400> 137 Cys Ile Tyr Thr Gly Ser Gly Ser Thr Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 138 <211> 17 <212> PRT <213> Artificial <220> <223> AS64183 CDR2 <400> 138 Ser Val Tyr Thr Asp Asp Asp Ser Thr Met Tyr Thr Asp Ser Val Lys 1 5 10 15 Gly <210> 139 <211> 17 <212> PRT <213> Artificial <220> <223> AS64207 CDR2 <400> 139 Thr Ile Val Ser Gly Gly Gly Thr Thr Tyr Tyr Ala Asp Ser Val Arg 1 5 10 15 Gly <210> 140 <211> 17 <212> PRT <213> Artificial <220> <223> AS64276 CDR2 <400> 140 Gln Ile Phe Thr Gly Arg Gly Thr Thr Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 141 <211> 16 <212> PRT <213> Artificial <220> <223> AS64336 CDR2 <400> 141 His Ile Phe Ser Asp Gly Ser Arg Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 142 <211> 20 <212> PRT <213> Artificial <220> <223> AS64346 CDR2 <400> 142 Thr Ile Gly Thr Gly Asp Ile Phe Asn Gly Ala Ala Tyr Tyr Val Asp 1 5 10 15 Ser Val Lys Gly 20 <210> 143 <211> 17 <212> PRT <213> Artificial <220> <223> AS64420 CDR2 <400> 143 Thr Ile Ser Thr Ala Gly Gly Ser Thr Trp Tyr Thr Asp Ser Val Lys 1 5 10 15 Gly <210> 144 <211> 17 <212> PRT <213> Artificial <220> <223> AS64473 CDR2 <400> 144 Cys Ile Tyr Thr Gly Ser Gly Thr Thr Gly Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 145 <211> 17 <212> PRT <213> Artificial <220> <223> AS64475 CDR2 <400> 145 Thr Ile Thr Ile Ser Gly Gly Ser Thr Trp Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 146 <211> 17 <212> PRT <213> Artificial <220> <223> AS64513 CDR2 <400> 146 Ala Ala Cys Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 147 <211> 17 <212> PRT <213> Artificial <220> <223> AS64562 CDR2 <400> 147 Val Ile His Thr Gly Gly Asp Thr Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 148 <211> 17 <212> PRT <213> Artificial <220> <223> AS64583 CDR2 <400> 148 Cys Thr Ser Trp Ala Gly Gly Arg Thr Tyr Thr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 149 <211> 17 <212> PRT <213> Artificial <220> <223> AS64594 CDR2 <400> 149 Leu Ile Tyr Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 150 <211> 17 <212> PRT <213> Artificial <220> <223> AS64605 CDR2 <400> 150 Arg Ile Tyr Thr Gly Thr Gly Ser Thr Trp Tyr Thr Asp Ser Val Lys 1 5 10 15 Gly <210> 151 <211> 20 <212> PRT <213> Artificial <220> <223> AS64606 CDR2 <400> 151 Ala Phe Tyr Leu Ile Tyr Thr Arg Gly Gly Ser Thr Tyr Tyr Ala Ser 1 5 10 15 Ser Val Lys Gly 20 <210> 152 <211> 17 <212> PRT <213> Artificial <220> <223> AS68121 CDR2 <400> 152 Thr Asp Tyr Ile Arg Phe Gly Arg Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 153 <211> 18 <212> PRT <213> Artificial <220> <223> AS68170 CDR2 <400> 153 Thr Ile Tyr Thr Gly Gly Gly Arg Asn Leu Tyr Tyr Ala Asp Ser Val 1 5 10 15 Lys Gly <210> 154 <211> 16 <212> PRT <213> Artificial <220> <223> AS63964 CDR2 <400> 154 Ala Ile Ser Gly Gly Arg Phe Thr Ala Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 155 <211> 17 <212> PRT <213> Artificial <220> <223> AS64116 CDR2 <400> 155 Gly Ile Ser Thr Gly Gly Gly Gly Thr Val Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 156 <211> 17 <212> PRT <213> Artificial <220> <223> AS68270 CDR2 <400> 156 Ala Ile Thr Ser Gly Thr Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys 1 5 10 15 Asp <210> 157 <211> 17 <212> PRT <213> Artificial <220> <223> AS68320 CDR2 <400> 157 Ala Ile Tyr Arg His Ser Gly Asn Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 158 <211> 17 <212> PRT <213> Artificial <220> <223> AS68351 CDR2 <400> 158 Gly Ile Ser Ala Ser Gly Gly Arg Thr Ser Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 159 <211> 17 <212> PRT <213> Artificial <220> <223> AS75378 CDR2 <400> 159 Ile Val Tyr Thr Gly Gly Arg Asp Thr Tyr Tyr Ala Ala Ser Val Lys 1 5 10 15 Gly <210> 160 <211> 16 <212> PRT <213> Artificial <220> <223> AS75383 CDR2 <400> 160 Thr Phe Asn Asn Arg Gly Val Ala Asn Tyr His Asp Ser Val Lys Gly 1 5 10 15 <210> 161 <211> 16 <212> PRT <213> Artificial <220> <223> AS75751 CDR2 <400> 161 Ala Ile Asp Thr Asp Gly Arg Thr Ser Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 162 <211> 16 <212> PRT <213> Artificial <220> <223> AS76422 CDR2 <400> 162 Thr Tyr Asn Asn Phe Gly Val Ala Asn Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <210> 163 <211> 17 <212> PRT <213> Artificial <220> <223> AS63930 CDR3 <400> 163 Glu Val Ala Asp Pro Thr Trp Gly Ser Arg Asp Gln Arg Arg Tyr Lys 1 5 10 15 Tyr <210> 164 <211> 9 <212> PRT <213> Artificial <220> <223> AS63932 CDR3 <400> 164 Arg Tyr Gly Ser Gly Asn Val Asn Tyr 1 5 <210> 165 <211> 21 <212> PRT <213> Artificial <220> <223> AS63951 CDR3 <400> 165 Arg Ser Phe Ser Ile Ala Val Cys Ala Thr Arg Ser Gly Ile Thr Arg 1 5 10 15 Ser Asn Phe Ala Tyr 20 <210> 166 <211> 20 <212> PRT <213> Artificial <220> <223> AS63984 CDR3 <400> 166 Gly Gly Pro Val Thr Asn Ala Pro Arg Trp Tyr Pro Leu Arg Pro Pro 1 5 10 15 Gly Tyr Asn Tyr 20 <210> 167 <211> 16 <212> PRT <213> Artificial <220> <223> AS63987 CDR3 <400> 167 Asp Arg Pro Phe Val Cys Asn Ile Ala Asn Met Arg Arg Ser Ser Asn 1 5 10 15 <210> 168 <211> 16 <212> PRT <213> Artificial <220> <223> AS63997 CDR3 <400> 168 Gly Tyr Leu Ser Gly Gly Ser Trp Asp Val Pro Gly Arg Tyr Asn Tyr 1 5 10 15 <210> 169 <211> 16 <212> PRT <213> Artificial <220> <223> AS64047 CDR3 <400> 169 Gly Phe Val Ser Gly Gly Arg Trp Asn Gln Ser Tyr Arg Tyr Lys Tyr 1 5 10 15 <210> 170 <211> 20 <212> PRT <213> Artificial <220> <223> AS64052 CDR3 <400> 170 Gly Gly Pro Pro Ala Asn Ala Asp Arg Trp Tyr Pro Leu Arg Pro Pro 1 5 10 15 Gly Tyr Asn Tyr 20 <210> 171 <211> 17 <212> PRT <213> Artificial <220> <223> AS64062 CDR3 <400> 171 Val Val Pro Arg Gly Gly Ser Cys Arg Leu Asp Glu Arg Gly Tyr Tyr 1 5 10 15 His <210> 172 <211> 14 <212> PRT <213> Artificial <220> <223> AS64072 CDR3 <400> 172 Glu Thr Thr Met Gly Trp Ala His Glu Arg Gly Tyr Arg Tyr 1 5 10 <210> 173 <211> 18 <212> PRT <213> Artificial <220> <223> AS64097 CDR3 <400> 173 Gly Pro Asp Leu Gly Gly Ser Trp Cys Arg Pro Val Glu Arg Ala Phe 1 5 10 15 Thr Tyr <210> 174 <211> 16 <212> PRT <213> Artificial <220> <223> AS64114 CDR3 <400> 174 Ala Ser Arg His Arg Leu Arg Leu Asn Asn Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 175 <211> 7 <212> PRT <213> Artificial <220> <223> AS64123 CDR3 <400> 175 Arg Phe Ser Glu Tyr Asn Tyr 1 5 <210> 176 <211> 20 <212> PRT <213> Artificial <220> <223> AS64130 CDR3 <400> 176 Gly Gly Pro Arg Thr Asn Asp Asp Arg Trp Tyr Pro Leu Arg Pro Pro 1 5 10 15 Gly Tyr Asn Tyr 20 <210> 177 <211> 17 <212> PRT <213> Artificial <220> <223> AS64137 CDR3 <400> 177 Ala Ala Ala Ser Thr Asp Val Arg Leu Leu Asp Pro Gly Asp Phe Ala 1 5 10 15 Tyr <210> 178 <211> 17 <212> PRT <213> Artificial <220> <223> AS64142 CDR3 <400> 178 Asp Thr Thr Ser Gly Tyr Cys Ser Gly Phe Trp Ser Thr Ser Arg Tyr 1 5 10 15 Ser <210> 179 <211> 17 <212> PRT <213> Artificial <220> <223> AS64154 CDR3 <400> 179 Ser Ser Pro Arg Trp Gly Gly Thr Cys Arg Arg Trp Ser Gln Tyr Asn 1 5 10 15 Tyr <210> 180 <211> 19 <212> PRT <213> Artificial <220> <223> AS64160 CDR3 <400> 180 Asp Phe Gly Arg Asp Lys Asn Tyr Leu Arg Pro Leu Leu Pro His Ala 1 5 10 15 Tyr Asn Tyr <210> 181 <211> 9 <212> PRT <213> Artificial <220> <223> AS64228 CDR3 <400> 181 Leu Cys Trp Arg Glu Asn Val Asn Tyr 1 5 <210> 182 <211> 15 <212> PRT <213> Artificial <220> <223> AS64300 CDR3 <400> 182 Phe Asn Tyr Gly Gly Ala Trp Tyr Glu Glu Arg Ser Tyr Lys Tyr 1 5 10 15 <210> 183 <211> 16 <212> PRT <213> Artificial <220> <223> AS64380 CDR3 <400> 183 Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 184 <211> 16 <212> PRT <213> Artificial <220> <223> AS64395 CDR3 <400> 184 Ala Ala Ser Thr Ala Val Arg Leu Leu Asp Pro Gly Asp Phe Ala Tyr 1 5 10 15 <210> 185 <211> 7 <212> PRT <213> Artificial <220> <223> AS64443 CDR3 <400> 185 Arg Phe Pro Ala Val Thr Tyr 1 5 <210> 186 <211> 16 <212> PRT <213> Artificial <220> <223> AS64511 CDR3 <400> 186 Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe Arg Ala 1 5 10 15 <210> 187 <211> 16 <212> PRT <213> Artificial <220> <223> AS64536 CDR3 <400> 187 Lys Trp Gly Leu Gly Gly Gly Gly Leu Lys Ser Asp Thr Tyr Met Tyr 1 5 10 15 <210> 188 <211> 20 <212> PRT <213> Artificial <220> <223> AS64597 CDR3 <400> 188 Gly Gly Pro Pro Thr Asp Gly Asp Arg Trp Tyr Pro Leu Arg Pro Pro 1 5 10 15 Gly Tyr Asn Tyr 20 <210> 189 <211> 10 <212> PRT <213> Artificial <220> <223> AS64617 CDR3 <400> 189 Ile Val Gly Arg Thr Cys Ser Leu Asn Tyr 1 5 10 <210> 190 <211> 16 <212> PRT <213> Artificial <220> <223> AS64634 CDR3 <400> 190 Arg Arg Trp Val Ser Gly Thr Cys Tyr Trp Asp Ser Asp Phe His Tyr 1 5 10 15 <210> 191 <211> 16 <212> PRT <213> Artificial <220> <223> AS69498 CDR3 <400> 191 Ser Ser Arg His Arg Leu Arg Leu Ser Asp Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 192 <211> 17 <212> PRT <213> Artificial <220> <223> AS69500 CDR3 <400> 192 Arg Ala Phe Gln Val Gly Tyr Cys Tyr Leu Arg Thr Asp Val Tyr Asn 1 5 10 15 Tyr <210> 193 <211> 16 <212> PRT <213> Artificial <220> <223> AS69527 CDR3 <400> 193 Arg Arg Trp Ala Cys Pro Arg Val Gly Ser Trp His Glu Phe Ala Tyr 1 5 10 15 <210> 194 <211> 16 <212> PRT <213> Artificial <220> <223> AS68280 CDR3 <400> 194 Ala Arg Gly Arg Phe Ile Asp Trp Thr Lys Ala Thr Gln Tyr Asp Tyr 1 5 10 15 <210> 195 <211> 20 <212> PRT <213> Artificial <220> <223> AS68355 CDR3 <400> 195 Ala Ile Val Gly Gly Phe Asn Ala Tyr Cys Ser Gly Gly Tyr Val Leu 1 5 10 15 Asp Phe Gly Ala 20 <210> 196 <211> 24 <212> PRT <213> Artificial <220> <223> AS69443 CDR3 <400> 196 Asp Phe Leu Thr Gly Phe Tyr Tyr Ser Asp Ser Pro His Pro Ala Pro 1 5 10 15 Cys Ser Ala Ser Asp Phe Gly Tyr 20 <210> 197 <211> 17 <212> PRT <213> Artificial <220> <223> AS75376 CDR3 <400> 197 Asp Pro Asn Pro Asp Tyr Met Leu Pro Phe Arg Pro Ser Arg Arg Ser 1 5 10 15 Trp <210> 198 <211> 14 <212> PRT <213> Artificial <220> <223> AS75387 CDR3 <400> 198 Asp Leu Arg Leu Pro Arg Ala Gly Gly Cys Ala Tyr Ser Tyr 1 5 10 <210> 199 <211> 13 <212> PRT <213> Artificial <220> <223> AS75695 CDR3 <400> 199 Asp Arg Arg Gly Arg Val Arg Arg Cys Glu Tyr Asn Ala 1 5 10 <210> 200 <211> 17 <212> PRT <213> Artificial <220> <223> AS76169 CDR3 <400> 200 Asp Arg Gly Gly Tyr Ala Asn Ser Cys Ala Val Ala Ala Arg Tyr Asp 1 5 10 15 Tyr <210> 201 <211> 16 <212> PRT <213> Artificial <220> <223> AS63931 CDR3 <400> 201 Gly Trp Leu Ser Gly Gly Ser Trp His Val Pro Gly Arg Tyr Asn Tyr 1 5 10 15 <210> 202 <211> 14 <212> PRT <213> Artificial <220> <223> AS63937 CDR3 <400> 202 Asp Ser Arg Arg Pro Cys Met Val Ala Ala Gly Tyr Thr Tyr 1 5 10 <210> 203 <211> 17 <212> PRT <213> Artificial <220> <223> AS63948 CDR3 <400> 203 Ala Ser Pro Arg Trp Gly Gly Thr Cys Arg Arg Trp Ser Glu Tyr Asn 1 5 10 15 Tyr <210> 204 <211> 15 <212> PRT <213> Artificial <220> <223> AS63956 CDR3 <400> 204 Tyr Gln Ala Lys Tyr Cys Ser Gly Pro Cys Ala Pro Pro Thr Asp 1 5 10 15 <210> 205 <211> 13 <212> PRT <213> Artificial <220> <223> AS63965 CDR3 <400> 205 Trp Cys Arg Glu Tyr Pro Gly Gly Ile Leu Asn Asn Gly 1 5 10 <210> 206 <211> 10 <212> PRT <213> Artificial <220> <223> AS63993 CDR3 <400> 206 Arg Thr Asp Ser Glu His Ala Phe Lys Phe 1 5 10 <210> 207 <211> 7 <212> PRT <213> Artificial <220> <223> AS63999 CDR3 <400> 207 Arg Tyr Thr Asp Tyr Asn Tyr 1 5 <210> 208 <211> 7 <212> PRT <213> Artificial <220> <223> AS64006 CDR3 <400> 208 Arg Ser Arg Thr Met Met Tyr 1 5 <210> 209 <211> 17 <212> PRT <213> Artificial <220> <223> AS64057 CDR3 <400> 209 Asp Thr Thr Ser Gly Ser Cys Ser Gly Phe Trp Ser Thr Ser Arg Tyr 1 5 10 15 Tyr <210> 210 <211> 14 <212> PRT <213> Artificial <220> <223> AS64060 CDR3 <400> 210 Asp Pro Thr Ile Gly Cys Pro Gln Thr Tyr Arg Tyr Asn Tyr 1 5 10 <210> 211 <211> 16 <212> PRT <213> Artificial <220> <223> AS64071 CDR3 <400> 211 Ser Thr Lys Asp Gln Phe Tyr Val Phe Asn Pro Ile Gly Tyr Asp Ser 1 5 10 15 <210> 212 <211> 21 <212> PRT <213> Artificial <220> <223> AS64093 CDR3 <400> 212 Arg Ser Phe Ser Ile Gly Val Cys Ala Thr Gln Ser Gly Ile Thr Trp 1 5 10 15 Ser Asn Phe Ala Tyr 20 <210> 213 <211> 10 <212> PRT <213> Artificial <220> <223> AS64118 CDR3 <400> 213 Thr Trp Asp Ser Ser Cys Arg Phe Gln Tyr 1 5 10 <210> 214 <211> 21 <212> PRT <213> Artificial <220> <223> AS64120 CDR3 <400> 214 Arg Ser Phe Ser Ile Ala Asp Cys Ala Thr Gln Ser Gly Ile Thr Arg 1 5 10 15 Ser Asn Phe Ala Tyr 20 <210> 215 <211> 19 <212> PRT <213> Artificial <220> <223> AS64124 CDR3 <400> 215 Gly Arg Pro Tyr Ser Leu Pro Leu Pro Leu Pro Leu Glu Ser Gly Ala 1 5 10 15 Tyr Arg Tyr <210> 216 <211> 13 <212> PRT <213> Artificial <220> <223> AS64135 CDR3 <400> 216 Asp Pro Met Gly Gly Ser Arg Thr Pro Cys Thr Ser Ala 1 5 10 <210> 217 <211> 17 <212> PRT <213> Artificial <220> <223> AS64163 CDR3 <400> 217 Asp Pro Ser Val Cys Pro Gly Gly Met Trp Tyr Ser Lys Glu Tyr Arg 1 5 10 15 Tyr <210> 218 <211> 17 <212> PRT <213> Artificial <220> <223> AS64182 CDR3 <400> 218 Ser Ser Pro His Trp Gly Gly Thr Cys Arg Arg Trp Ser Glu Tyr Lys 1 5 10 15 Tyr <210> 219 <211> 13 <212> PRT <213> Artificial <220> <223> AS64183 CDR3 <400> 219 Asp Leu Ser Gly Gly Pro Ala Gly Cys Gly Tyr Thr His 1 5 10 <210> 220 <211> 20 <212> PRT <213> Artificial <220> <223> AS64207 CDR3 <400> 220 Gly Gly Pro Pro Thr Asn Gly Ala Lys Trp Tyr Pro Leu Arg Pro Pro 1 5 10 15 Gly Tyr Asn Tyr 20 <210> 221 <211> 13 <212> PRT <213> Artificial <220> <223> AS64276 CDR3 <400> 221 Ser Leu Gly Pro Gly Arg Gly Ala Cys Gly Tyr Asn Tyr 1 5 10 <210> 222 <211> 13 <212> PRT <213> Artificial <220> <223> AS64336 CDR3 <400> 222 Arg Thr Gly Trp Ala Pro Arg Cys Ala Val Pro Gly Tyr 1 5 10 <210> 223 <211> 18 <212> PRT <213> Artificial <220> <223> AS64346 CDR3 <400> 223 Val Gln Ser Lys Ser Ser Asn Tyr Val Leu Arg Asp Ala Ser Thr Tyr 1 5 10 15 Asn Tyr <210> 224 <211> 7 <212> PRT <213> Artificial <220> <223> AS64420 CDR3 <400> 224 Arg Ser Arg Tyr Ala Thr Tyr 1 5 <210> 225 <211> 17 <212> PRT <213> Artificial <220> <223> AS64473 CDR3 <400> 225 Ser Ser Pro Gln Trp Gly Gly Thr Cys Arg Arg Trp Ser Glu Tyr Asn 1 5 10 15 Tyr <210> 226 <211> 7 <212> PRT <213> Artificial <220> <223> AS64475 CDR3 <400> 226 Arg Asp Thr Ala Arg Thr Tyr 1 5 <210> 227 <211> 18 <212> PRT <213> Artificial <220> <223> AS64513 CDR3 <400> 227 Gly Tyr Tyr Ser Gly Ser Gly Pro Gly Tyr Leu Leu Pro Trp Arg Tyr 1 5 10 15 Asn Tyr <210> 228 <211> 22 <212> PRT <213> Artificial <220> <223> AS64562 CDR3 <400> 228 Gly Phe Arg Pro Arg Gly Gly Gly Tyr Thr Gly Asp Val Leu Ala Gln 1 5 10 15 Ala Ala Ala Tyr Asn Tyr 20 <210> 229 <211> 17 <212> PRT <213> Artificial <220> <223> AS64583 CDR3 <400> 229 Lys Ala His Pro Asp Cys Ser Gly Asp Trp Ser Pro Ser Gly Tyr Glu 1 5 10 15 Tyr <210> 230 <211> 7 <212> PRT <213> Artificial <220> <223> AS64594 CDR3 <400> 230 Arg Thr Gln Thr Arg Asn Tyr 1 5 <210> 231 <211> 22 <212> PRT <213> Artificial <220> <223> AS64605 CDR3 <400> 231 Arg Ser Asn Ser Tyr Ser Tyr Ser Ser Cys Asp Tyr Gly Pro Leu Thr 1 5 10 15 Arg Gly Gly Tyr Asn Phe 20 <210> 232 <211> 7 <212> PRT <213> Artificial <220> <223> AS64606 CDR3 <400> 232 Arg Leu Asp Glu Lys Met Tyr 1 5 <210> 233 <211> 21 <212> PRT <213> Artificial <220> <223> AS68121 CDR3 <400> 233 Asp Pro Gly Ser Arg Thr Asp Asp Ser Cys Gly Thr Ser Tyr Asn Lys 1 5 10 15 Gly Asn Phe Gly Tyr 20 <210> 234 <211> 20 <212> PRT <213> Artificial <220> <223> AS68170 CDR3 <400> 234 Ala Ser Asp Val Ala Val Gly Val Asn Ser Cys Gly Gly Arg Thr Ala 1 5 10 15 Gly Phe Asp Ala 20 <210> 235 <211> 17 <212> PRT <213> Artificial <220> <223> AS63964 CDR3 <400> 235 Glu Val Val Asp Pro Thr Trp Gly Ser Arg Asp Gln Arg Arg Tyr Lys 1 5 10 15 Tyr <210> 236 <211> 17 <212> PRT <213> Artificial <220> <223> AS64116 CDR3 <400> 236 Asp Arg Trp Asn Ser Phe Ala Asn Cys Gly Ala Trp Gly Arg Tyr Thr 1 5 10 15 Tyr <210> 237 <211> 18 <212> PRT <213> Artificial <220> <223> AS68270 CDR3 <400> 237 Ala Ser Gly Trp Ile Val Pro Ser Arg Ser Leu Thr Ala Asn Leu Tyr 1 5 10 15 Arg Tyr <210> 238 <211> 16 <212> PRT <213> Artificial <220> <223> AS68320 CDR3 <400> 238 Gly Arg Ala Gly Pro Trp Ala Leu Met Arg Pro Thr Glu Phe Gly Tyr 1 5 10 15 <210> 239 <211> 14 <212> PRT <213> Artificial <220> <223> AS68351 CDR3 <400> 239 Gly Ala Val Arg Leu Ser Thr Ser Ser Val Arg Asp Ser Ser 1 5 10 <210> 240 <211> 7 <212> PRT <213> Artificial <220> <223> AS75378 CDR3 <400> 240 Arg Ser Tyr Glu Tyr Thr Tyr 1 5 <210> 241 <211> 16 <212> PRT <213> Artificial <220> <223> AS75383 CDR3 <400> 241 Asp Arg Arg Tyr Gly Arg Gln Trp Tyr Gln Pro Cys Glu Trp Asn Thr 1 5 10 15 <210> 242 <211> 20 <212> PRT <213> Artificial <220> <223> AS75751 CDR3 <400> 242 Gly Phe Gly Tyr Met Asn Val Ile Gln Ala Leu Asn Gly Met Arg Gln 1 5 10 15 Asn Pro Asp Tyr 20 <210> 243 <211> 16 <212> PRT <213> Artificial <220> <223> AS76422 CDR3 <400> 243 Asp Arg Arg Asp Gly Arg Arg Trp Ser Gln Pro Cys Glu Trp Asn Thr 1 5 10 15 <210> 244 <211> 10 <212> PRT <213> Artificial <220> <223> AS64380VH4 CDR1 <400> 244 Gly Asn Thr Tyr Ser Ser Asn Tyr Met Gly 1 5 10 <210> 245 <211> 10 <212> PRT <213> Artificial <220> <223> AS64380VH5 CDR1 <400> 245 Gly Asn Thr Tyr Ser Ser Asn Tyr Met Gly 1 5 10 <210> 246 <211> 10 <212> PRT <213> Artificial <220> <223> AS64380VH6 CDR1 <400> 246 Gly Asn Thr Tyr Ser Ser Asn Tyr Met Gly 1 5 10 <210> 247 <211> 10 <212> PRT <213> Artificial <220> <223> AS64380VH7 CDR1 <400> 247 Gly Asn Thr Tyr Ser Ser Asn Tyr Met Gly 1 5 10 <210> 248 <211> 10 <212> PRT <213> Artificial <220> <223> AS64511VH4 CDR1 <400> 248 Arg Ala Thr Tyr Ser Thr Asn Tyr Ile Ser 1 5 10 <210> 249 <211> 10 <212> PRT <213> Artificial <220> <223> AS64511VH5 CDR1 <400> 249 Arg Ala Thr Tyr Ser Thr Asn Tyr Ile Ser 1 5 10 <210> 250 <211> 10 <212> PRT <213> Artificial <220> <223> AS64511VH6 CDR1 <400> 250 Arg Ala Thr Tyr Ser Thr Asn Tyr Ile Ser 1 5 10 <210> 251 <211> 10 <212> PRT <213> Artificial <220> <223> AS63997VH4 CDR1 <400> 251 Phe Ser Gly Tyr Gly Val Ser Thr Met Ala 1 5 10 <210> 252 <211> 10 <212> PRT <213> Artificial <220> <223> AS63997VH5 CDR1 <400> 252 Phe Ser Gly Tyr Gly Val Ser Thr Met Ala 1 5 10 <210> 253 <211> 10 <212> PRT <213> Artificial <220> <223> AS63997VH6 CDR1 <400> 253 Phe Ser Gly Tyr Gly Val Ser Thr Met Ala 1 5 10 <210> 254 <211> 17 <212> PRT <213> Artificial <220> <223> AS64380VH4 CDR2 <400> 254 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val Arg 1 5 10 15 Gly <210> 255 <211> 17 <212> PRT <213> Artificial <220> <223> AS64380VH5 CDR2 <400> 255 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val Arg 1 5 10 15 Gly <210> 256 <211> 17 <212> PRT <213> Artificial <220> <223> AS64380VH6 CDR2 <400> 256 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val Arg 1 5 10 15 Gly <210> 257 <211> 17 <212> PRT <213> Artificial <220> <223> AS64380VH7 CDR2 <400> 257 Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val Arg 1 5 10 15 Gly <210> 258 <211> 17 <212> PRT <213> Artificial <220> <223> AS64511VH4 CDR2 <400> 258 Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 259 <211> 17 <212> PRT <213> Artificial <220> <223> AS64511VH5 CDR2 <400> 259 Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 260 <211> 17 <212> PRT <213> Artificial <220> <223> AS64511VH6 CDR2 <400> 260 Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 261 <211> 17 <212> PRT <213> Artificial <220> <223> AS63997VH4 CDR2 <400> 261 Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 262 <211> 17 <212> PRT <213> Artificial <220> <223> AS63997VH5 CDR2 <400> 262 Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 263 <211> 17 <212> PRT <213> Artificial <220> <223> AS63997VH6 CDR2 <400> 263 Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 264 <211> 16 <212> PRT <213> Artificial <220> <223> AS64380VH4 CDR3 <400> 264 Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 265 <211> 16 <212> PRT <213> Artificial <220> <223> AS64380VH5 CDR3 <400> 265 Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 266 <211> 16 <212> PRT <213> Artificial <220> <223> AS64380VH6 CDR3 <400> 266 Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 267 <211> 16 <212> PRT <213> Artificial <220> <223> AS64380VH7 CDR3 <400> 267 Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr Asp Tyr 1 5 10 15 <210> 268 <211> 16 <212> PRT <213> Artificial <220> <223> AS64511VH4 CDR3 <400> 268 Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe Arg Ala 1 5 10 15 <210> 269 <211> 16 <212> PRT <213> Artificial <220> <223> AS64511VH5 CDR3 <400> 269 Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe Arg Ala 1 5 10 15 <210> 270 <211> 16 <212> PRT <213> Artificial <220> <223> AS64511VH6 CDR3 <400> 270 Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe Arg Ala 1 5 10 15 <210> 271 <211> 16 <212> PRT <213> Artificial <220> <223> AS63997VH4 CDR3 <400> 271 Gly Tyr Leu Ser Gly Gly Ser Trp Asp Val Pro Gly Arg Tyr Asn Tyr 1 5 10 15 <210> 272 <211> 16 <212> PRT <213> Artificial <220> <223> AS63997VH5 CDR3 <400> 272 Gly Tyr Leu Ser Gly Gly Ser Trp Asp Val Pro Gly Arg Tyr Asn Tyr 1 5 10 15 <210> 273 <211> 16 <212> PRT <213> Artificial <220> <223> AS63997VH6 CDR3 <400> 273 Gly Tyr Leu Ser Gly Gly Ser Trp Asp Val Pro Gly Arg Tyr Asn Tyr 1 5 10 15 <210> 274 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63930 <400> 274 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Gly Asn 20 25 30 Tyr Met Ala Trp Phe Arg Gln Ala Pro Gly Asn Glu Arg Glu Gly Val 35 40 45 Ala Val Val Tyr Asn Ile Asp Gly Gly Gly Arg Phe Thr Thr Tyr Ala 50 55 60 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Gly Asn Asp Lys Asn 65 70 75 80 Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Ser Gly Met 85 90 95 Tyr Tyr Cys Ala Ala Glu Val Ala Asp Pro Thr Trp Gly Ser Arg Asp 100 105 110 Gln Arg Arg Tyr Lys Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 275 <211> 118 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63932 <400> 275 Gln Val Gln Leu Glu Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Thr Tyr Gly Ser Thr 20 25 30 Phe Met Gly Trp Phe Arg Gln Asn Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Val Ile Tyr Thr Gly Gly Gly Ser Thr Trp Tyr Ala Ser Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Tyr Gly Ser Gly Asn Val Asn Tyr Trp Gly Gln Gly Thr 100 105 110 Gln Val Thr Val Ser Ser 115 <210> 276 <211> 130 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63951 <400> 276 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ala Cys Glu Thr Ser Arg Asp Ile Tyr Gly Asn Asn 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ser Ile Tyr Pro Ala Gly Gly Arg Pro Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Ser Phe Ser Ile Ala Val Cys Ala Thr Arg Ser Gly Ile 100 105 110 Thr Arg Ser Asn Phe Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 115 120 125 Ser Ser 130 <210> 277 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63984 <400> 277 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Ile Val Ser Gly Gly Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Gly Pro Val Thr Asn Ala Pro Arg Trp Tyr Pro Leu Arg 100 105 110 Pro Pro Gly Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 278 <211> 122 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63987 <400> 278 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Thr Gly Tyr Arg Asn Cys Met Ala 20 25 30 Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala Val Ile 35 40 45 Tyr Thr Pro Ser Gly Ile Thr Asp Tyr Ala Ser Ser Val Lys Gly Arg 50 55 60 Phe Thr Ile Ser Gln Asn Asn Ala Arg Asn Thr Gln Tyr Leu Gln Met 65 70 75 80 Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala Ala Asp 85 90 95 Arg Pro Phe Val Cys Asn Ile Ala Asn Met Arg Arg Ser Ser Asn Trp 100 105 110 Gly Arg Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 279 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63997 <400> 279 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Phe Ser Gly Tyr Gly Val Ser 20 25 30 Thr Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Thr Val Phe 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Val Gly Tyr Leu Ser Gly Gly Ser Trp Asp Val Pro Gly Arg Tyr 100 105 110 Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 280 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64047 <400> 280 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gln Tyr Val Tyr Arg Trp Asp 20 25 30 Leu Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45 Ala Ala Val Tyr Thr Gly Asp Gly Ile Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ser Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Met Tyr Phe Cys 85 90 95 Ala Ala Gly Phe Val Ser Gly Gly Arg Trp Asn Gln Ser Tyr Arg Tyr 100 105 110 Lys Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 281 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64052 <400> 281 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Arg Ser Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Ile 35 40 45 Ala Thr Ile His Ser Gly Val Ala Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Gly Pro Pro Ala Asn Ala Asp Arg Trp Tyr Pro Leu Arg 100 105 110 Pro Pro Gly Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 282 <211> 127 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64062 <400> 282 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Val Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Ser Pro Tyr Ser Ser Ser 20 25 30 Arg Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly 35 40 45 Val Ala Ala Leu Tyr Thr Gly Gly Gly Ser Thr Ser Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr 85 90 95 Cys Ala Ala Val Val Pro Arg Gly Gly Ser Cys Arg Leu Asp Glu Arg 100 105 110 Gly Tyr Tyr His Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 283 <211> 123 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64072 <400> 283 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Phe Leu Arg Leu Ser Cys Ala Leu Ser Gly Tyr Ser Tyr Tyr Ile Asn 20 25 30 Leu Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45 Ala Ala His Gly Pro Val Ser Gly Thr Ala Tyr Tyr Thr Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Pro Gly Lys Asn Thr Met Tyr 65 70 75 80 Leu Gln Met Phe Ser Leu Gln Pro Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95 Ala Ala Glu Thr Thr Met Gly Trp Ala His Glu Arg Gly Tyr Arg Tyr 100 105 110 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 284 <211> 127 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64097 <400> 284 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Tyr Thr Tyr Ser Arg Asn 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Asn Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Leu 50 55 60 Glu Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Met Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Pro Asp Leu Gly Gly Ser Trp Cys Arg Pro Val Glu Arg 100 105 110 Ala Phe Thr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 285 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64114 <400> 285 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Thr Leu Ser Cys Glu Ala Ser Gly Asn Thr Tyr Ser Thr Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ala Ser Arg His Arg Leu Arg Leu Asn Asn Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 286 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64123 <400> 286 Gln Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Thr Ser Asn 20 25 30 Trp Leu Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Ile Ile Tyr Thr Gly Ser Gly Ser Thr His Tyr Arg Ser Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Phe Ser Glu Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 287 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64130 <400> 287 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Arg Ser Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Ala Pro Ala Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Ile Asp Ser Arg Gly Thr Ile Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Glu Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Gly Pro Arg Thr Asn Asp Asp Arg Trp Tyr Pro Leu Arg 100 105 110 Pro Pro Gly Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 288 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64137 <400> 288 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Tyr Ser Thr Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Leu Val Thr Trp Val Glu Arg Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Arg Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ala Ala Ala Ser Thr Asp Val Arg Leu Leu Asp Pro Gly Asp 100 105 110 Phe Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 289 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64142 <400> 289 Gln Val His Leu Met Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Asp Arg Asn 20 25 30 Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ser Cys Ile Asp Trp Thr Gly Ala Asn Ile Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Met Tyr Tyr Cys 85 90 95 Ala Ala Asp Thr Thr Ser Gly Tyr Cys Ser Gly Phe Trp Ser Thr Ser 100 105 110 Arg Tyr Ser Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 290 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64154 <400> 290 Gln Val Gln Leu Lys Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Tyr Thr Tyr Arg Tyr Leu 20 25 30 Tyr Met Gly Trp Phe Arg Gln Thr Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Cys Ile Tyr Thr Gly Ser Gly Ser Thr Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Pro Arg Trp Gly Gly Thr Cys Arg Arg Trp Ser Gln 100 105 110 Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 291 <211> 128 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64160 <400> 291 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Val Tyr Thr Ser Ser Ser Tyr 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Met Cys Phe Gly Gly Leu Val Thr His Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asp Phe Gly Arg Asp Lys Asn Tyr Leu Arg Pro Leu Leu Pro 100 105 110 His Ala Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 292 <211> 117 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64228 <400> 292 Gln Val Gln Leu Lys Glu Ser Gly Gly Gly Ser Ile Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Val Ser Tyr Asn Arg Cys 20 25 30 Ser Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Gly Arg Glu Leu Val 35 40 45 Ser Arg Ile Gln Pro Gly Gly Asn Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Val Ser Gln Asp Asn Ala Lys Asn Thr Val Ser Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Asn 85 90 95 Ala Leu Cys Trp Arg Glu Asn Val Asn Tyr Trp Gly Gln Gly Thr Gln 100 105 110 Val Thr Val Ser Ser 115 <210> 293 <211> 124 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64300 <400> 293 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Asp Ile Tyr Asn Leu Met 20 25 30 Ser Met Ala Trp Phe Arg Arg Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Tyr Ile Asn Thr Ile Ile Gly Asn Thr Tyr Tyr Thr Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Phe Asn Tyr Gly Gly Ala Trp Tyr Glu Glu Arg Ser Tyr Lys 100 105 110 Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 294 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64380 <400> 294 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Thr Leu Ser Cys Glu Ala Ser Gly Asn Thr Tyr Ser Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 295 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64395 <400> 295 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Tyr Ser Thr Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Leu Val Thr Trp Ala Glu Arg Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Arg Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Gly Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ala Ala Ser Thr Ala Val Arg Leu Leu Asp Pro Gly Asp Phe 100 105 110 Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 296 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64443 <400> 296 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Asp Ser Ser Val 20 25 30 Tyr Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Ile Ile Tyr Thr Gly Gly Glu Ser Thr His Tyr Arg Ser Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Gln Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Phe Pro Ala Val Thr Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 297 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64511 <400> 297 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Ala Thr Tyr Ser Thr Asn 20 25 30 Tyr Ile Ser Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45 Ala Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe 100 105 110 Arg Ala Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 298 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64536 <400> 298 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Tyr Thr Asp Asn Phe Val 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Leu Ile Tyr Pro Gly Gly Gly Ser Thr Tyr Tyr Ala Ser Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Gly Thr Val His 65 70 75 80 Leu Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Lys Trp Gly Leu Gly Gly Gly Gly Leu Lys Ser Asp Thr Tyr 100 105 110 Met Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 299 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64597 <400> 299 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Arg Val Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Thr Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Ile Asp Ser Gly Val Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser His Asn Asn Ala Lys Asn Thr Ile Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Gly Pro Pro Thr Asp Gly Asp Arg Trp Tyr Pro Leu Arg 100 105 110 Pro Pro Gly Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 300 <211> 117 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64617 <400> 300 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Asp Arg Cys Ser 20 25 30 Met Ala Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val Ser 35 40 45 Arg Ile Ser Thr Ser Gly Phe Thr Asn Tyr Ala Ala Ser Val Lys Gly 50 55 60 Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln 65 70 75 80 Met Asn Ser Leu Asn Pro Gly Asp Thr Gly Met Tyr Tyr Cys Ala Ile 85 90 95 Ile Val Gly Arg Thr Cys Ser Leu Asn Tyr Trp Gly Asn Gly Ile Leu 100 105 110 Val Thr Val Ser Ser 115 <210> 301 <211> 127 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64634 <400> 301 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ser Phe Arg Gly Asp 20 25 30 Phe Met Cys Met Gly Trp Phe Arg Gln Thr Pro Gly Lys Gly Arg Glu 35 40 45 Gly Val Ala Val Phe Tyr Pro Gly Gly Gly Ser Thr Asn Tyr Ala Asp 50 55 60 Ser Ala Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr 65 70 75 80 Met Tyr Leu Gln Met Asn Thr Leu Lys Pro Glu Asp Thr Ala Met Tyr 85 90 95 Tyr Cys Ala Ala Arg Arg Trp Val Ser Gly Thr Cys Tyr Trp Asp Ser 100 105 110 Asp Phe His Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 302 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS69498 <400> 302 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Gly Asn Thr Tyr Ser Ser Asn 20 25 30 Tyr Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Ile Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Arg His Arg Leu Arg Leu Ser Asp Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 303 <211> 127 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS69500 <400> 303 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Asp Arg Tyr Thr Tyr Ser Ser Ala 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ser Ile Phe Thr Gly Thr Gly Gly Ser Thr Tyr Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr 85 90 95 Cys Ala Ala Arg Ala Phe Gln Val Gly Tyr Cys Tyr Leu Arg Thr Asp 100 105 110 Val Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 304 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS69527 <400> 304 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Arg Tyr Thr Phe Ser Ser Thr 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Ala Ile Tyr Thr Asp Asp Gly Ser Thr Trp Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Arg Trp Ala Cys Pro Arg Val Gly Ser Trp His Glu Phe 100 105 110 Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 305 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68280 <400> 305 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val His Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Tyr Ser Ser Asn 20 25 30 Tyr Leu Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Asp Trp Val 35 40 45 Ala Ala Ile Ser Thr Gly Asp Gly Ala Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Leu Glu Asp Ser Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ala Arg Gly Arg Phe Ile Asp Trp Thr Lys Ala Thr Gln Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 306 <211> 128 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68355 <400> 306 Gln Met Gln Leu Val Glu Ser Gly Gly Asp Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Gly Val 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Asp Ser Asp Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Ala Ala Ile Val Gly Gly Phe Asn Ala Tyr Cys Ser Gly Gly Tyr Val 100 105 110 Leu Asp Phe Gly Ala Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 307 <211> 132 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS69443 <400> 307 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Phe Thr Phe Asp Asp Ser 20 25 30 Asp Met Ala Trp Tyr Arg Gln Ala Pro Gly Asp Gly Cys Asp Leu Val 35 40 45 Ser Thr Ile Ser Ser Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met His Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Ala Asp Phe Leu Thr Gly Phe Tyr Tyr Ser Asp Ser Pro His Pro Ala 100 105 110 Pro Cys Ser Ala Ser Asp Phe Gly Tyr Trp Gly Gln Gly Thr Gln Val 115 120 125 Thr Val Ser Ser 130 <210> 308 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS75376 <400> 308 Gln Val Gln Leu Lys Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser His 20 25 30 Ser Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Val Ile Tyr Thr Gly Asp Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asp Pro Asn Pro Asp Tyr Met Leu Pro Phe Arg Pro Ser Arg 100 105 110 Arg Ser Trp Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 309 <211> 123 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS75387 <400> 309 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Pro Tyr Ser Ser Pro 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Leu Val Ala Tyr Thr Gly Gly Asp Ile Gln Tyr Leu Thr Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asp Leu Arg Leu Pro Arg Ala Gly Gly Cys Ala Tyr Ser Tyr 100 105 110 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 310 <211> 121 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS75695 <400> 310 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Thr Val Ser Ala Tyr 20 25 30 Cys Met Gly Trp Phe Arg Gln Val Leu Gly Lys Gly Arg Glu Arg Ile 35 40 45 Ala Phe Ile Asp Ala Gly Gly Ala Thr Ile Tyr Ala Asp Pro Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Val 85 90 95 Ala Asp Arg Arg Gly Arg Val Arg Arg Cys Glu Tyr Asn Ala Trp Gly 100 105 110 Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 311 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS76169 <400> 311 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ile Tyr Ser Ser Phe 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Val Val 35 40 45 Ala Tyr Ile Arg Asp Asn Ile Met Thr Ser Tyr Thr Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Lys Arg Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Gly Leu Lys Pro Glu Asp Thr Gly Met Tyr Tyr Cys Ala 85 90 95 Val Asp Arg Gly Gly Tyr Ala Asn Ser Cys Ala Val Ala Ala Arg Tyr 100 105 110 Asp Tyr Trp Gly Arg Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 312 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63931 <400> 312 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Phe Ser Gly Tyr Gly Val Ser 20 25 30 Thr Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Arg Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Trp Leu Ser Gly Gly Ser Trp His Val Pro Gly Arg Tyr 100 105 110 Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 313 <211> 123 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63937 <400> 313 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Ile Ser Ser Arg 20 25 30 Pro Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Cys Ile His Thr Gly Leu Gly Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Val Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asp Ser Arg Arg Pro Cys Met Val Ala Ala Gly Tyr Thr Tyr 100 105 110 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 314 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63948 <400> 314 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Tyr Thr Tyr Arg Tyr Leu 20 25 30 Tyr Met Gly Trp Phe Arg Gln Thr Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Cys Ile Tyr Thr Gly Ser Gly Ser Thr Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Glu Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ala Ser Pro Arg Trp Gly Gly Thr Cys Arg Arg Trp Ser Glu 100 105 110 Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 315 <211> 123 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63956 <400> 315 Gln Val His Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Tyr Ser Asn Cys 20 25 30 Cys Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Ala Arg Glu Leu Val 35 40 45 Ser Leu Ile Asn Ser Ser Gly Gly Thr Tyr Tyr Ala Asp Ser Val Arg 50 55 60 Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Ala Tyr Gln Ala Lys Tyr Cys Ser Gly Pro Cys Ala Pro Pro Thr Asp 100 105 110 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 316 <211> 121 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63965 <400> 316 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Ser Ser Gly Ser Cys 20 25 30 Arg Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val 35 40 45 Ser Lys Val Ile Ser Asp Gly Thr Thr Val Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Leu Ser Gln Gly Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Ser Ser Leu Leu Pro Glu Asp Thr Ala Met Tyr Tyr Cys Asn 85 90 95 Ala Trp Cys Arg Glu Tyr Pro Gly Gly Ile Leu Asn Asn Gly Trp Gly 100 105 110 Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 317 <211> 118 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63993 <400> 317 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Val Ser Gly Phe Thr Phe Asp Asp Leu 20 25 30 Val Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Gln Leu Val 35 40 45 Ser Leu Val Ala Thr Ala Gly Asn Ser Val Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Leu Ser Arg Asp Asn Ala His Ser Thr Ala Tyr Leu 65 70 75 80 Gln Met Asn Gly Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Ala Arg Thr Asp Ser Glu His Ala Phe Lys Phe Trp Gly Gln Gly Thr 100 105 110 Gln Val Thr Val Ser Ser 115 <210> 318 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63999 <400> 318 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser Asn 20 25 30 Trp Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Glu Val 35 40 45 Ala Ile Ile Tyr Thr Gly Gly Ile Ser Thr His Tyr Arg Ser Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Tyr Thr Asp Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 319 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64006 <400> 319 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Val Ser Gly Tyr Thr Gly Asp Thr Thr 20 25 30 Tyr Ile Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Leu Ile Tyr Thr Ser Gly Thr Ser Glu Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ile Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Gly Ala Arg Ser Arg Thr Met Met Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 320 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64057 <400> 320 Gln Val Gln Leu Glu Glu Ser Gly Gly Gly Leu Val Gln Thr Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Asp Arg Asn 20 25 30 Ala Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ser Cys Ile Ser Trp Thr Gly Ala Asn Ile Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Met Tyr Tyr Cys 85 90 95 Ala Ala Asp Thr Thr Ser Gly Ser Cys Ser Gly Phe Trp Ser Thr Ser 100 105 110 Arg Tyr Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 321 <211> 122 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64060 <400> 321 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Ser Thr Tyr Cys Thr Tyr 20 25 30 Arg Met Ser Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45 Ala Val Ile Asp Ser Gly Gly Ser Thr Ser Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Lys 85 90 95 Thr Asp Pro Thr Ile Gly Cys Pro Gln Thr Tyr Arg Tyr Asn Tyr Trp 100 105 110 Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 322 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64071 <400> 322 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Asn Thr Tyr Arg Leu Asn 20 25 30 Ser Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Phe Ile Val Met Ile Arg Gly Thr Thr Tyr Tyr Gly Ala Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Gln Thr Thr Val Tyr 65 70 75 80 Leu Gln Met Ser Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Thr Lys Asp Gln Phe Tyr Val Phe Asn Pro Ile Gly Tyr 100 105 110 Asp Ser Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 323 <211> 130 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64093 <400> 323 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Thr Ser Arg Tyr Ile Tyr Gly Asn Asn 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ser Ile Tyr Pro Ala Gly Gly Arg Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Ile Asp Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Ser Phe Ser Ile Gly Val Cys Ala Thr Gln Ser Gly Ile 100 105 110 Thr Trp Ser Asn Phe Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 115 120 125 Ser Ser 130 <210> 324 <211> 117 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64118 <400> 324 Gln Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ala Cys 20 25 30 Arg Met Ala Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val 35 40 45 Ser Phe Ile Asn Ser Ala Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Ala Ile Ser Arg Asp Asn Ala Lys Thr Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ala Leu Lys Ala Glu Asp Thr Ala Ile Tyr Tyr Cys Asn 85 90 95 Thr Trp Asp Ser Ser Cys Arg Phe Gln Tyr Trp Gly Gln Gly Thr Gln 100 105 110 Val Thr Val Ser Ser 115 <210> 325 <211> 130 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64120 <400> 325 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Thr Ser Arg Tyr Ile Tyr Gly Asn Asn 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ser Ile Tyr Pro Ala Gly Gly Arg Pro Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Ser Phe Ser Ile Ala Asp Cys Ala Thr Gln Ser Gly Ile 100 105 110 Thr Arg Ser Asn Phe Ala Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 115 120 125 Ser Ser 130 <210> 326 <211> 128 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64124 <400> 326 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Thr Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Thr Tyr Thr Pro Ser Asn Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Ala Thr Ile Gly Gly Thr Thr Arg Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Gly Ala Lys Asn Thr Ile Tyr 65 70 75 80 Leu Gln Met Asn Gly Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Arg Pro Tyr Ser Leu Pro Leu Pro Leu Pro Leu Glu Ser 100 105 110 Gly Ala Tyr Arg Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 327 <211> 121 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64135 <400> 327 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Thr Ser Thr Tyr Cys Arg Tyr 20 25 30 Tyr Met Arg Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Phe Val 35 40 45 Ser Ala Met Gln Pro Asp Gly Thr Thr Ser Tyr Ser Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Met Ser Gln Asp Arg Ala Asn Asn Met Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Met Tyr Tyr Cys Lys 85 90 95 Arg Asp Pro Met Gly Gly Ser Arg Thr Pro Cys Thr Ser Ala Trp Gly 100 105 110 Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 328 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64163 <400> 328 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Arg Tyr Arg Trp Asn 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Ser Thr Gly Ser Gly Ser Thr Tyr Tyr Ala Gly Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Met Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Ala Asp Pro Ser Val Cys Pro Gly Gly Met Trp Tyr Ser Lys Glu Tyr 100 105 110 Arg Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 329 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64182 <400> 329 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Gln Thr Ser Arg Tyr Leu 20 25 30 Tyr Met Gly Trp Phe Arg Gln Thr Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Cys Ile Tyr Thr Gly Ser Gly Ser Thr Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Thr Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Pro His Trp Gly Gly Thr Cys Arg Arg Trp Ser Glu 100 105 110 Tyr Lys Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 330 <211> 122 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64183 <400> 330 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly His Thr Tyr Ser Ala Asn 20 25 30 Cys Met Ala Trp Phe Arg Arg Ala Pro Gly Lys Glu Arg Glu Trp Val 35 40 45 Ala Ser Val Tyr Thr Asp Asp Asp Ser Thr Met Tyr Thr Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Phe Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Gly Met Tyr Ile Cys 85 90 95 Ala Ala Asp Leu Ser Gly Gly Pro Ala Gly Cys Gly Tyr Thr His Trp 100 105 110 Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 331 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64207 <400> 331 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Ser Ser Asn 20 25 30 Phe Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Ile Val Ser Gly Gly Gly Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Gly Pro Pro Thr Asn Gly Ala Lys Trp Tyr Pro Leu Arg 100 105 110 Pro Pro Gly Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 332 <211> 122 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64276 <400> 332 Gln Val His Leu Met Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Ser Leu Ser Cys Val Val Ser Gly Tyr Thr Gly Ser Ser Arg 20 25 30 Cys Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Ala Val 35 40 45 Ala Gln Ile Phe Thr Gly Arg Gly Thr Thr Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Arg Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Ala Ser Leu Gly Pro Gly Arg Gly Ala Cys Gly Tyr Asn Tyr Trp 100 105 110 Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 333 <211> 121 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64336 <400> 333 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Thr Ser Gly Arg Thr Tyr Ser Ser Cys 20 25 30 Ser Met Gly Trp Tyr Arg Gln Ala Pro Gly Lys Glu Arg Glu Leu Val 35 40 45 Ser His Ile Phe Ser Asp Gly Ser Arg Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Asn 85 90 95 Arg Arg Thr Gly Trp Ala Pro Arg Cys Ala Val Pro Gly Tyr Trp Gly 100 105 110 Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 334 <211> 127 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64346 <400> 334 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Phe Met Gly 20 25 30 Trp Phe Arg Gln Ala Pro Gln Lys Glu Arg Glu Trp Val Ala Thr Ile 35 40 45 Gly Thr Gly Asp Ile Phe Asn Gly Ala Ala Tyr Tyr Val Asp Ser Val 50 55 60 Lys Gly Arg Phe Ala Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Val Tyr Val Cys 85 90 95 Ala Ala Val Gln Ser Lys Ser Ser Asn Tyr Val Leu Arg Asp Ala Ser 100 105 110 Thr Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 335 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64420 <400> 335 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Glu Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asp Thr Ser Arg Ser Val 20 25 30 Trp Met Gly Trp Ala Arg Gln Val Pro Gly Lys Glu Arg Glu Val Val 35 40 45 Ala Thr Ile Ser Thr Ala Gly Gly Ser Thr Trp Tyr Thr Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Ala Arg Ser Arg Tyr Ala Thr Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 336 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64473 <400> 336 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Tyr Thr Tyr Arg Tyr Leu 20 25 30 Tyr Met Ala Trp Phe Arg Gln Thr Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Cys Ile Tyr Thr Gly Ser Gly Thr Thr Gly Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Asn Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Pro Gln Trp Gly Gly Thr Cys Arg Arg Trp Ser Glu 100 105 110 Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 337 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64475 <400> 337 Gln Val Gln Leu Gln Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Trp Ser Arg Asn 20 25 30 Trp Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Phe 35 40 45 Ala Thr Ile Thr Ile Ser Gly Gly Ser Thr Trp Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Leu Asp Asn Ala Gly Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Asp Thr Ala Arg Thr Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 338 <211> 127 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64513 <400> 338 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Asp Tyr Pro Tyr Ile Asp Asn 20 25 30 Cys Met Gly Trp Phe Arg Gln Gly Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ala Cys Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Asp Val Tyr Tyr Cys 85 90 95 Ala Thr Gly Tyr Tyr Ser Gly Ser Gly Pro Gly Tyr Leu Leu Pro Trp 100 105 110 Arg Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 339 <211> 131 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64562 <400> 339 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Ala Arg Arg Asp 20 25 30 Phe Met Ala Trp Phe Arg Gln Val Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Val Ile His Thr Gly Gly Asp Thr Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Gln Asn Ile Met Asn 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Phe Arg Pro Arg Gly Gly Gly Tyr Thr Gly Asp Val Leu 100 105 110 Ala Gln Ala Ala Ala Tyr Asn Tyr Trp Gly Gln Gly Thr Gln Val Thr 115 120 125 Val Ser Ser 130 <210> 340 <211> 126 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64583 <400> 340 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Leu Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Ile Ala Val Tyr 20 25 30 Thr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Ile 35 40 45 Ser Cys Thr Ser Trp Ala Gly Gly Arg Thr Tyr Thr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Lys Ala His Pro Asp Cys Ser Gly Asp Trp Ser Pro Ser Gly 100 105 110 Tyr Glu Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 341 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64594 <400> 341 Gln Val His Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Asn Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Leu Ile Tyr Thr Gly Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ser Val Arg Thr Gln Thr Arg Asn Tyr Trp Gly Gln Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 342 <211> 131 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64605 <400> 342 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Arg Tyr Pro Tyr Ser Ser Ile 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Ser Glu Gly Val 35 40 45 Ala Arg Ile Tyr Thr Gly Thr Gly Ser Thr Trp Tyr Thr Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ala Arg Asp Asn Ala Gln Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Ser Asn Ser Tyr Ser Tyr Ser Ser Cys Asp Tyr Gly Pro 100 105 110 Leu Thr Arg Gly Gly Tyr Asn Phe Trp Gly Gln Gly Thr Gln Val Thr 115 120 125 Val Ser Ser 130 <210> 343 <211> 115 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64606 <400> 343 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Tyr Thr Ser Arg Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Leu Ile Tyr Thr Arg Gly Gly Ser Thr Tyr Tyr Ala Ser Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Ser Ala Lys Lys Thr Tyr Leu 65 70 75 80 Gln Met Asn Ser Val Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Leu Arg Leu Asp Glu Lys Met Tyr Trp Gly Gln Gly Thr Gln Val Thr 100 105 110 Val Ser Ser 115 <210> 344 <211> 133 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68121 <400> 344 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Asp Ala Ser Gly Tyr Thr Tyr Ser Arg Asn 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Phe Tyr Thr Asp Tyr Ile Arg Phe Gly Arg Thr Tyr Tyr Ala 50 55 60 Asp Ser Val Lys Gly Arg Phe Thr Ile Phe Gln Asp Asn Ala Lys Asn 65 70 75 80 Thr Val Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met 85 90 95 Tyr Tyr Cys Ala Ala Asp Pro Gly Ser Arg Thr Asp Asp Ser Cys Gly 100 105 110 Thr Ser Tyr Asn Lys Gly Asn Phe Gly Tyr Trp Gly Gln Gly Thr Gln 115 120 125 Val Thr Val Ser Ser 130 <210> 345 <211> 130 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68170 <400> 345 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Tyr Thr Tyr Arg Ser Asn 20 25 30 Cys Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Thr Ile Tyr Thr Gly Gly Gly Arg Asn Leu Tyr Tyr Ala Asp Ser 50 55 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Ser Ala Arg Tyr Tyr 85 90 95 Cys Ala Ala Ala Ser Asp Val Ala Val Gly Val Asn Ser Cys Gly Gly 100 105 110 Arg Thr Ala Gly Phe Asp Ala Trp Gly Gln Gly Thr Gln Val Thr Val 115 120 125 Ser Ser 130 <210> 346 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS63964 <400> 346 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ser Ala Ser Gly Tyr Thr Tyr Ser Tyr Asn 20 25 30 Asn Met Gly Trp Phe Arg Gln Ala Pro Gly Asn Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Ser Gly Gly Arg Phe Thr Ala Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Glu Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Asn Leu Lys Pro Glu Asp Thr Gly Met Tyr Tyr Cys Ala 85 90 95 Ala Glu Val Val Asp Pro Thr Trp Gly Ser Arg Asp Gln Arg Arg Tyr 100 105 110 Lys Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 347 <211> 124 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS64116 <400> 347 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ile Tyr Ser Cys Val 20 25 30 Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val Ala Gly 35 40 45 Ile Ser Thr Gly Gly Gly Gly Thr Val Tyr Ala Asp Ser Val Lys Gly 50 55 60 Gln Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Val Tyr Leu Gln 65 70 75 80 Met Asp Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala Ala 85 90 95 Asp Arg Trp Asn Ser Phe Ala Asn Cys Gly Ala Trp Gly Arg Tyr Thr 100 105 110 Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 348 <211> 130 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68270 <400> 348 Gln Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Pro Ser Ser Thr Tyr 20 25 30 Tyr Met Leu Ser Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg 35 40 45 Glu Gly Val Ala Ala Ile Thr Ser Gly Thr Gly Ser Thr Ser Tyr Ala 50 55 60 Asp Ser Val Lys Asp Arg Phe Thr Ile Ser Lys Asp Tyr Ala Asn Asn 65 70 75 80 Thr Leu Tyr Leu His Ile Asn Asn Leu Lys Pro Glu Asp Thr Ala Met 85 90 95 Tyr Tyr Cys Ala Ala Ala Ser Gly Trp Ile Val Pro Ser Arg Ser Leu 100 105 110 Thr Ala Asn Leu Tyr Arg Tyr Trp Gly Gln Gly Thr Gln Val Thr Val 115 120 125 Ser Ser 130 <210> 349 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68320 <400> 349 Gln Val His Leu Val Glu Ser Gly Gly Asp Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Tyr Asn Thr Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ala Ile Tyr Arg His Ser Gly Asn Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Tyr Ala Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Arg Ala Gly Pro Trp Ala Leu Met Arg Pro Thr Glu Phe 100 105 110 Gly Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 125 <210> 350 <211> 123 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS68351 <400> 350 Gln Val Gln Leu Glu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asp Thr Phe Arg Ala Tyr 20 25 30 Tyr Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Phe Glu Trp Val 35 40 45 Ser Gly Ile Ser Ala Ser Gly Gly Arg Thr Ser Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Leu Asn Ser Leu Ser Thr Glu Asp Thr Gly Met Tyr Tyr Cys 85 90 95 Val Lys Gly Ala Val Arg Leu Ser Thr Ser Ser Val Arg Asp Ser Ser 100 105 110 Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 351 <211> 116 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS75378 <400> 351 Gln Val Gln Leu Glu Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Val Ser Gly Asn Thr Arg Ser Thr Thr 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly Val 35 40 45 Ala Ile Val Tyr Thr Gly Gly Arg Asp Thr Tyr Tyr Ala Ala Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Thr Thr Ile Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Glu Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Arg Ser Tyr Glu Tyr Thr Tyr Trp Gly Arg Gly Thr Gln Val 100 105 110 Thr Val Ser Ser 115 <210> 352 <211> 124 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS75383 <400> 352 Glu Val Gln Leu Ala Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Thr Phe Ser Ser Tyr 20 25 30 Cys Leu Gly Trp Phe Arg Gln Ala Pro Gly Lys Gln Arg Gln Gly Val 35 40 45 Ala Thr Phe Asn Asn Arg Gly Val Ala Asn Tyr His Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ala Ser Val Asp Asn Ala Lys Asn Thr Leu Leu Leu 65 70 75 80 Gln Met Asn Ser Leu Glu Pro Asp Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Ala Asp Arg Arg Tyr Gly Arg Gln Trp Tyr Gln Pro Cys Glu Trp Asn 100 105 110 Thr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 353 <211> 129 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS75751 <400> 353 Gln Val Arg Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Tyr Phe Tyr Asn Thr Tyr 20 25 30 Tyr Phe Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Glu Arg Glu Gly 35 40 45 Val Ala Ala Ile Asp Thr Asp Gly Arg Thr Ser Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Lys Asp Asn Ala Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Lys Pro Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Phe Gly Tyr Met Asn Val Ile Gln Ala Leu Asn Gly Met 100 105 110 Arg Gln Asn Pro Asp Tyr Trp Gly Gln Gly Thr Gln Val Thr Val Ser 115 120 125 Ser <210> 354 <211> 124 <212> PRT <213> Artificial <220> <223> Anti-DLL3 camel sdAb amino acid sequence AS76422 <400> 354 Gln Val Lys Leu Val Glu Ser Gly Gly Gly Ser Val Gln Ala Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Ala Gly Asn 20 25 30 Cys Leu Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Gly Val 35 40 45 Val Thr Tyr Asn Asn Phe Gly Val Ala Asn Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Val Ser Gln Asp Asn Ala Lys Asn Thr Leu Leu Leu 65 70 75 80 Gln Met Asn Ser Leu Glu Pro Glu Asp Thr Ala Met Tyr Tyr Cys Ala 85 90 95 Ala Asp Arg Arg Asp Gly Arg Arg Trp Ser Gln Pro Cys Glu Trp Asn 100 105 110 Thr Trp Gly Gln Gly Thr Gln Val Thr Val Ser Ser 115 120 <210> 355 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64380VH4 <400> 355 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Thr Tyr Ser Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 356 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64380VH5 <400> 356 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Thr Tyr Ser Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 357 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64380VH6 <400> 357 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Thr Tyr Ser Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Ser Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 358 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64380VH7 <400> 358 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Asn Thr Tyr Ser Ser Asn 20 25 30 Tyr Met Gly Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Glu Val 35 40 45 Ala Val Ile Tyr Thr Arg Gly Gly His Thr Tyr Tyr Val Asp Ser Val 50 55 60 Arg Gly Arg Phe Thr Ile Ser Gln Asp Asn Ala Lys Asn Ser Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Ser Ser Arg His Arg Leu Gly Leu Asn Asn Pro Arg Asp Tyr 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 359 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64511VH4 <400> 359 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Ala Thr Tyr Ser Thr Asn 20 25 30 Tyr Ile Ser Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Ala Val 35 40 45 Ala Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe 100 105 110 Arg Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 360 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64511VH5 <400> 360 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Ala Thr Tyr Ser Thr Asn 20 25 30 Tyr Ile Ser Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Ala Val 35 40 45 Ala Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe 100 105 110 Arg Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 361 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS64511VH6 <400> 361 Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Arg Ala Thr Tyr Ser Thr Asn 20 25 30 Tyr Ile Ser Trp Phe Arg Gln Ala Pro Gly Lys Gly Arg Glu Ala Val 35 40 45 Ala Thr Ile Thr Thr Gly Asp Gly Glu Thr Ala Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Asn Leu Arg Ile Gly Gly Asp Trp Phe Asp Gly Arg Asp Phe 100 105 110 Arg Ala Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 362 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS63931VH4 <400> 362 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Phe Ser Gly Tyr Gly Val Ser 20 25 30 Thr Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Gly Val 35 40 45 Ala Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Ala Gly Trp Leu Ser Gly Gly Ser Trp His Val Pro Gly Arg Tyr 100 105 110 Asn Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 363 <211> 125 <212> PRT <213> Artificial <220> <223> Anti-DLL3 humanized sdAb amino acid sequence AS63931VH5 <400> 363 Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Gly Ser Phe Ser Gly Tyr Gly Val Ser 20 25 30 Thr Met Ala Trp Phe Arg Gln Ala Pro Gly Lys Gly Leu Glu Gly Val 35 40 45 Ala Ala Ile Thr Val Gly Ser Gly Asn Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Thr Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Val Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Met Tyr Tyr Cys 85 90 95 Ala Ala Gly Trp Leu Ser Gly Gly Ser Trp His Val Pro Gly Arg Tyr 100 105 110 Asn Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 <210> 364 ...

Claims

1. 1. A chimeric antigen receptor (CAR) comprising a DLL3-binding domain, wherein the DLL3-binding domain comprises or is derived from a single domain antibody (sdAb) or a single chain variable fragment (scFv).

2. The CAR of claim 1, wherein the sdAb comprises a polypeptide comprising: CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81, or a variant thereof comprising up to about three amino acid substitutions in the CDR1; CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162, or a variant thereof comprising up to about three amino acid substitutions in the CDR2; and CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243, or a variant thereof comprising up to about three amino acid substitutions in the CDR3.

3. 3. The CAR of claim 1 or 2, wherein the sdAb comprises a polypeptide comprising CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1 to 81, CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82 to 162, and CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163 to 243, or a variant of the polypeptide comprising up to about three amino acid substitutions in the CDR1, the CDR2, and the CDR3.

4. the sdAb (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions. The CAR according to any one of claims 1 to 3, comprising a polypeptide comprising any one of:

5. the sdAb (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 The CAR according to any one of claims 1 to 4, comprising a polypeptide comprising any one of:

6. The CAR of any one of claims 1 to 5, wherein the sdAb is a camelid sdAb raised against human or rhesus DLL3.

7. The CAR of any one of claims 1 to 6, wherein the sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274 to 354.

8. The CAR of any one of claims 1 to 7, wherein the sdAb is humanized through CDR grafting.

9. The CAR of any one of claims 1 to 8, wherein the humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355 to 367.

10. The CAR according to any one of claims 1 to 9, wherein the scFv comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH of the scFv comprises CDR1 shown in SEQ ID NO: 498 or 504 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, CDR2 shown in SEQ ID NO: 499 or 505 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and CDR3 shown in SEQ ID NO: 500 or 506 or a variant thereof comprising up to about three amino acid substitutions in the CDR3; and the VL of the scFv comprises CDR1 shown in SEQ ID NO: 495 or 501 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, CDR2 shown in SEQ ID NO: 496 or 502 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and CDR3 shown in SEQ ID NO: 497 or 503 or a variant thereof comprising up to about three amino acid substitutions in the CDR3.

11. The CAR according to any one of claims 1 to 10, wherein the VH of the scFv comprises CDR1 set forth in SEQ ID NO:498, CDR2 set forth in SEQ ID NO:499, and CDR3 set forth in SEQ ID NO:500, and the VL of the scFv comprises CDR1 set forth in SEQ ID NO:495, CDR2 set forth in SEQ ID NO:496, and CDR3 set forth in SEQ ID NO:497; or the VH of the scFv comprises CDR1 set forth in SEQ ID NO:504, CDR2 set forth in SEQ ID NO:505, and CDR3 set forth in SEQ ID NO:506, and the VL of the scFv comprises CDR1 set forth in SEQ ID NO:501, CDR2 set forth in SEQ ID NO:502, and CDR3 set forth in SEQ ID NO:

503.

12. The CAR according to any one of claims 1 to 11, wherein the VH of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 508, and the VL of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 507; or the VH of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO: 510, and the VL of the scFv comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:

509.

13. The CAR of any one of claims 1 to 12, wherein the scFv is obtained from a synthetic human Fab phage library.

14. The CAR according to any one of claims 1 to 13, wherein the DLL3 is human or rhesus DLL3.

15. The CAR according to any one of claims 1 to 14, comprising, from the N-terminus to the C-terminus, a signal peptide, the DLL3-binding domain, a hinge domain, a transmembrane domain, and an intracellular signaling domain.

16. The CAR according to any one of claims 1 to 15, wherein the intracellular signaling domain is derived from CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, or CD66d.

17. The CAR according to any one of claims 1 to 16, wherein the intracellular signaling domain further comprises an intracellular costimulatory sequence.

18. The CAR according to any one of claims 1 to 17, wherein the intracellular costimulatory sequence is derived from a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD40, PD-1, LFA-1, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, TNFRSF9, TNFRSF4, TNFRSF8, CD40LG, ITGB2, KLRC2, TNFRSF18, TNFRSF14, HAVCR1, LGALS9, DAP10, DAP12, CD83, a CD83 ligand, and a combination thereof.

19. The CAR according to any one of claims 1 to 18, comprising an amino acid sequence having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 476 to 484, 485 to 494, and 515 to 516.

20. The CAR of any one of claims 1 to 19, wherein the DLL3-binding domain comprises two sdAbs linked to each other.

21. 21. The CAR of any one of claims 1 to 20, wherein each of the sdAbs independently comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO: 356 or SEQ ID NO:

366.

22. The CAR according to any one of claims 1 to 21, comprising an amino acid sequence having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 518 to 520.

23. The CAR according to any one of claims 1 to 22, comprising the amino acid sequence of SEQ ID NO:

520.

24. 1. A DLL3 binding protein comprising a single domain antibody (sdAb) portion that specifically binds to DLL3, wherein the sdAb portion comprises a polypeptide comprising: a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81, or a variant thereof comprising up to about three amino acid substitutions in said CDR1; a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162, or a variant thereof comprising up to about three amino acid substitutions in said CDR2; and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243, or a variant thereof comprising up to about three amino acid substitutions in said CDR3.

25. 25. The DLL3 binding protein of claim 24, wherein the sdAb portion comprises a polypeptide comprising a CDR1 comprising the amino acid sequence of any one of SEQ ID NOs: 1-81, a CDR2 comprising the amino acid sequence of any one of SEQ ID NOs: 82-162, and a CDR3 comprising the amino acid sequence of any one of SEQ ID NOs: 163-243, or a variant of said polypeptide comprising up to about three amino acid substitutions in said CDR1, said CDR2, and said CDR3.

26. the sdAb portion (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 87 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168 or a variant thereof comprising up to about three amino acid substitutions; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 102 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183 or a variant thereof comprising up to about three amino acid substitutions; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 105 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186 or a variant thereof comprising up to about three amino acid substitutions; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 108 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189 or a variant thereof comprising up to about three amino acid substitutions; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 115 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196 or a variant thereof comprising up to about three amino acid substitutions; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 120 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201 or a variant thereof comprising up to about three amino acid substitutions; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 88 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169 or a variant thereof comprising up to about three amino acid substitutions; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about three amino acid substitutions; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising up to about three amino acid substitutions; CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising up to about three amino acid substitutions; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising up to about three amino acid substitutions.

26. The DLL3 binding protein of claim 24 or 25, comprising a polypeptide comprising any one of:

27. the sdAb portion (1) CDR1 comprising the amino acid sequence of SEQ ID NO: 6; CDR2 comprising the amino acid sequence of SEQ ID NO: 87; and CDR3 comprising the amino acid sequence of SEQ ID NO: 168; (2) CDR1 comprising the amino acid sequence of SEQ ID NO: 21; CDR2 comprising the amino acid sequence of SEQ ID NO: 102; and CDR3 comprising the amino acid sequence of SEQ ID NO: 183; (3) CDR1 comprising the amino acid sequence of SEQ ID NO: 24; CDR2 comprising the amino acid sequence of SEQ ID NO: 105; and CDR3 comprising the amino acid sequence of SEQ ID NO: 186; (4) CDR1 comprising the amino acid sequence of SEQ ID NO: 27; CDR2 comprising the amino acid sequence of SEQ ID NO: 108; and CDR3 comprising the amino acid sequence of SEQ ID NO: 189; (5) CDR1 comprising the amino acid sequence of SEQ ID NO: 34; CDR2 comprising the amino acid sequence of SEQ ID NO: 115; and CDR3 comprising the amino acid sequence of SEQ ID NO: 196; (6) CDR1 comprising the amino acid sequence of SEQ ID NO: 39; CDR2 comprising the amino acid sequence of SEQ ID NO: 120; and CDR3 comprising the amino acid sequence of SEQ ID NO: 201; (7) CDR1 comprising the amino acid sequence of SEQ ID NO: 7; CDR2 comprising the amino acid sequence of SEQ ID NO: 88; and CDR3 comprising the amino acid sequence of SEQ ID NO: 169; (8) CDR1 comprising the amino acid sequence of SEQ ID NO: 8; CDR2 comprising the amino acid sequence of SEQ ID NO: 89; and CDR3 comprising the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 comprising the amino acid sequence of SEQ ID NO: 9; CDR2 comprising the amino acid sequence of SEQ ID NO: 90; and CDR3 comprising the amino acid sequence of SEQ ID NO: 171 27. The DLL3 binding protein of any one of claims 24 to 26, comprising a polypeptide comprising any one of:

28. 28. A DLL3 binding protein according to any one of claims 24 to 27, wherein the sdAb moiety is a camelid sdAb raised against human or rhesus DLL3.

29. 29. The DLL3 binding protein of any one of claims 24 to 28, wherein the sdAb portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 274 to 354.

30. 30. The DLL3 binding protein of any one of claims 24 to 29, wherein the sdAb portion is humanized through CDR grafting.

31. 31. The DLL3 binding protein of any one of claims 24 to 30, wherein the humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 355 to 367.

32. 32. The DLL3 binding protein of any one of claims 24 to 31, wherein the DLL3 is human or rhesus DLL3.

33. 1. A DLL3 binding protein comprising a single chain variable fragment (scFv) portion that specifically binds to DLL3, the scFv portion comprising a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH of the scFv portion comprises a CDR1 set forth in SEQ ID NO:498 or 504 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, a CDR2 set forth in SEQ ID NO:499 or 505 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and a CDR3 set forth in SEQ ID NO:500 or 506 or a CDR1 set forth in SEQ ID NO: 495 or 501 or a variant thereof comprising up to about three amino acid substitutions in the CDR1, a CDR2 set forth in SEQ ID NO: 496 or 502 or a variant thereof comprising up to about three amino acid substitutions in the CDR2, and a CDR3 set forth in SEQ ID NO: 497 or 503 or a variant thereof comprising up to about three amino acid substitutions in the CDR3.

34. 34. The DLL3 binding protein of claim 33, wherein the VH of the scFv portion comprises CDR1 set forth in SEQ ID NO:498, CDR2 set forth in SEQ ID NO:499, and CDR3 set forth in SEQ ID NO:500, and the VL of the scFv portion comprises CDR1 set forth in SEQ ID NO:495, CDR2 set forth in SEQ ID NO:496, and CDR3 set forth in SEQ ID NO:497; or the VH of the scFv portion comprises CDR1 set forth in SEQ ID NO:504, CDR2 set forth in SEQ ID NO:505, and CDR3 set forth in SEQ ID NO:506, and the VL of the scFv portion comprises CDR1 set forth in SEQ ID NO:501, CDR2 set forth in SEQ ID NO:502, and CDR3 set forth in SEQ ID NO:

503.

35. 35. The DLL3 binding protein of claim 33 or 34, wherein the VH of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:508 and the VL of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:507; or the VH of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:510 and the VL of the scFv portion comprises an amino acid sequence having at least about 95% sequence identity to the amino acid sequence set forth in SEQ ID NO:

509.

36. 36. The DLL3 binding protein of any one of claims 33 to 35, wherein the scFv portion is obtained from a synthetic human Fab phage library.

37. 37. The DLL3 binding protein of any one of claims 33 to 36, wherein the DLL3 is human or rhesus DLL3.

38. An isolated nucleic acid molecule encoding the CAR of any one of claims 1 to 23 or the DLL3-binding protein of any one of claims 24 to 32 or 33 to 37.

39. 39. The isolated nucleic acid molecule of claim 38, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 368-448, which encodes a camelid single domain antibody (sdAb).

40. 40. The isolated nucleic acid molecule of claim 38 or 39, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 449 to 461, which encodes a humanized camelid sdAb.

41. 41. The isolated nucleic acid molecule of any one of claims 38 to 40, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 511 to 514, encoding the VL or VH domain of a human scFv.

42. 42. The isolated nucleic acid molecule of any one of claims 38 to 41, further comprising a polynucleotide sequence encoding a chimeric switch receptor (CSR) or a dominant negative receptor (DNR).

43. 43. The isolated nucleic acid molecule of any one of claims 38 to 42, further comprising a polynucleotide sequence encoding a PD-1 dominant negative receptor (PD-1 DNR), a PD-1 chimeric switch receptor (PD-1 CSR), or a TGF-β dominant negative receptor (TGF-β DNR).

44. 44. The isolated nucleic acid molecule of any one of claims 38-43, wherein the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:523; and the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:

524.

45. 45. The isolated nucleic acid molecule of any one of claims 38 to 44, wherein the TGF-β DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:

529.

46. 46. ​​The isolated nucleic acid molecule of any one of claims 38-45, wherein the polynucleotide sequence encoding the PD-1 DNR, the PD-1 CSR, or the TGF-β DNR is linked to the polynucleotide sequence encoding the CAR via a polynucleotide sequence encoding a 2A self-cleaving peptide.

47. 47. The isolated nucleic acid molecule of any one of claims 38 to 46, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide.

48. 48. The isolated nucleic acid molecule of any one of claims 38 to 47, comprising, in the 5' to 3' direction, a polynucleotide sequence encoding the CAR, a polynucleotide sequence encoding the 2A self-cleaving peptide, and a polynucleotide sequence encoding the PD-1 DNR, the PD-1 CSR, or the TGF-β DNR.

49. 49. The isolated nucleic acid molecule of any one of claims 38 to 48, which encodes a peptide having at least about 95% sequence identity to SEQ ID NO: 521 or 522, or encodes a peptide having at least about 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 525 to 528.

50. 50. An expression vector comprising the isolated nucleic acid molecule of any one of claims 38 to 49.

51. A modified immune cell comprising the isolated nucleic acid molecule of any one of claims 38 to 49 or the expression vector of claim 50.

52. 52. The modified immune cell of claim 51, wherein the modified immune cell is selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and nature killer cells.

53. A modified immune cell expressing the CAR according to any one of claims 1 to 23.

54. 54. The modified immune cell of claim 53, which also expresses a CSR or a DNR.

55. 55. The modified immune cell of claim 53 or 54, wherein the CSR is a PD-1 CSR and the DNR is a PD-1 DNR or a TGF-β DNR.

56. 56. The modified immune cell of any one of claims 53-55, wherein the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:523; and the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:

524.

57. 57. The modified immune cell of any one of claims 53-56, wherein the TGF-β DNR comprises an amino acid sequence having at least about 95% sequence identity to SEQ ID NO:

529.

58. The modified immune cell of any one of claims 53 to 57, wherein the CAR and the CSR, or the CAR and the DNR, are simultaneously expressed via a 2A self-cleaving peptide.

59. 59. The modified immune cell of any one of claims 53 to 58, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide.

60. 60. The modified immune cell of any one of claims 53 to 59, wherein the modified immune cell expresses the CAR and the PD-1 CSR.

61. The modified immune cell of any one of claims 53 to 60, which expresses the CAR and the TGF-β DNR, and is stimulated by a cell expressing DLL3 in the presence of TGF-β.

62. The modified immune cell of any one of claims 53 to 61, wherein the modified immune cell is selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and nature killer cells.

63. 63. A pharmaceutical composition comprising the CAR of any one of claims 1 to 23, the DLL3 binding protein of any one of claims 24 to 32 or 33 to 37, the expression vector of claim 50, or the modified immune cell of any one of claims 51 to 62, and a physiologically acceptable excipient.

64. 63. A method for treating a DLL3-associated disorder in a subject, comprising administering to the subject a therapeutically effective amount of the modified immune cells of any one of claims 51-62, or a therapeutically effective amount of the pharmaceutical composition of claim 63.

65. 65. The method of claim 64, wherein the DLL3-associated disorder is a cancer selected from the group consisting of lung cancer, melanoma, breast cancer, prostate cancer, colon cancer, renal cell cancer, ovarian cancer, neuroblastoma, rhabdomyosarcoma, leukemia, and lymphoma.

66. 66. The method of claim 65, wherein the DLL3-associated disorder is small cell lung cancer.

67. 67. The method of any one of claims 64 to 66, wherein the cancer expresses DLL3 and PD-L1.

68. 68. The method of any one of claims 64 to 67, wherein the cancer has a higher TGF-β expression level compared to corresponding normal tissue.

69. Use of a CAR according to any one of claims 1 to 23, a DLL3-binding protein according to any one of claims 24 to 32 or 33 to 37, an expression vector according to claim 50, or a modified immune cell according to any one of claims 51 to 62, for the preparation of a medicament for treating a DLL3-associated disorder.

70. 63. A medicament for use in the treatment of a DLL3-associated disorder, the medicament comprising a CAR of any one of claims 1 to 23, a DLL3-binding protein of any one of claims 24 to 32 or claims 33 to 37, an expression vector of claim 50, or a modified immune cell of any one of claims 51 to 62.

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