Anti-DLL3 chimeric antigen receptor and its use

Abstract

To provide improved therapeutic approaches for treating small cell lung cancer (SCLC) and other DLL3 (delta-like ligand 3)-expressing cancers.SOLUTION: Provided is a chimeric antigen receptor (CAR) comprising a DLL3-binding domain, the DLL3-binding domain comprising or being derived from a single domain antibody (sdAb) or a single chain variable fragment (scFv).SELECTED DRAWING: None

Description

[Technical Field]

[0001] This invention relates to a chimeric antigen receptor (CAR) that targets DLL3 and a DLL3-specific binding protein. The invention also relates to nucleic acid sequences encoding a CAR or binding protein, modified immune cells expressing a CAR, and their use for treating DLL3-related disorders. [Background technology]

[0002] Advances in cellular immunotherapy present promising approaches for treating various tumors. One such therapy involves genetically modifying 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 function of T cells in a conventional form. 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 responsiveness, and proliferation, and can therefore act as "living drugs" to eradicate targeted tumor cells. In principle, any antigen (e.g., cell surface molecules) can be targeted by these CAR-T cells. CAR-T cell therapy can neutralize tolerance to autoantigens and provide treatment independent of the patient's MHC status. Recent trials have demonstrated remarkable clinical responses in leukemia and lymphoma patients using T cells modified to express a CD19-targeting chimeric antigen receptor.

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

[0004] The DLL3 (delta-like ligand 3) protein has been found to be clinically associated with various proliferative disorders, including neuroendocrine-characterized tumors such as small cell lung cancer (SCLC). SCLC, originating from neuroendocrine progenitor cells, accounts for 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 (Non-patent Literature 1)). This is because it is highly invasive, approximately two-thirds of patients have metastatic disease at diagnosis, and is highly refractory to conventional treatment (e.g., platinum-based chemotherapy).

[0005] Improved therapeutic approaches are needed to treat SCLC and other DLL3-expressing cancers. [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 [Overview of the project]

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

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

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

[0010] In some embodiments, sdAb is as follows: (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 87 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 168 or its variants containing up to approximately 3 amino acid substitutions; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 102 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 183 or its variants containing up to approximately 3 amino acid substitutions; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 105 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 186 or its variants containing up to approximately 3 amino acid substitutions; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 108 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 189 or its variants containing up to approximately 3 amino acid substitutions; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 115 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 196 or its variants containing up to approximately 3 amino acid substitutions; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 120 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 201 or its variants containing up to approximately 3 amino acid substitutions; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 88 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 169 or its variants containing up to approximately 3 amino acid substitutions; (8) A CDR1 comprising the amino acid sequence of SEQ ID NO: 8 or a variant thereof comprising at most about 3 amino acid substitutions; a CDR2 comprising the amino acid sequence of SEQ ID NO: 89 or a variant thereof comprising at most about 3 amino acid substitutions; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising at most about 3 amino acid substitutions; or (9) A CDR1 comprising the amino acid sequence of SEQ ID NO: 9 or a variant thereof comprising at most about 3 amino acid substitutions; a CDR2 comprising the amino acid sequence of SEQ ID NO: 90 or a variant thereof comprising at most about 3 amino acid substitutions; and a CDR3 comprising the amino acid sequence of SEQ ID NO: 171 or a variant thereof comprising at most about 3 amino acid substitutions comprises a polypeptide comprising any one of them.

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

[0012] In some embodiments, the sdAb is a camel sdAb produced against human or rhesus monkey DLL3.

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

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

[0015] In some embodiments, the humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity with any one of the amino acid sequences 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 includes CDR1 or a variant thereof containing up to about three amino acid substitutions in CDR1, as shown in SEQ ID NO: 498 or 504, CDR2 or a variant thereof containing up to about three amino acid substitutions in CDR2, as shown in SEQ ID NO: 499 or 505, and CDR3 or a variant thereof containing up to about three amino acid substitutions in CDR3, as shown in SEQ ID NO: 500 or 506, and the VL domain of the scFv includes CDR1 or a variant thereof containing up to about three amino acid substitutions in CDR1, as shown in SEQ ID NO: 495 or 501, CDR2 or a variant thereof containing up to about three amino acid substitutions in CDR2, as shown in SEQ ID NO: 496 or 502, and CDR3 or a variant thereof containing up to about three amino acid substitutions in CDR3, as shown in SEQ ID NO: 497 or 503.

[0017] In some embodiments, the VH domain of scFv includes 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 domain of scFv includes 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 domain of scFv includes 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 domain of scFv includes CDR1 shown in SEQ ID NO: 501, CDR2 shown in SEQ ID NO: 502, and CDR3 shown in SEQ ID NO: 503.

[0018] In some embodiments, the VH domain of scFv contains 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 domain of scFv contains 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 domain of scFv contains 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 domain of scFv contains an amino acid sequence having at least about 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 509.

[0019] In some embodiments, scFv is obtained from a synthetic human Fab phage library.

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

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

[0022] In some embodiments, the intracellular signaling domain is derived from CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, or CD66d.

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

[0024] In some embodiments, the intracellular co-stimulatory sequence is derived from a co-stimulatory 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, the ligand of CD83, and combinations thereof.

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

[0026] In some embodiments, the DLL3 binding domain includes two sdAbs linked together.

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

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

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

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

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

[0032] In some embodiments, the sdAb portion is as follows: (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 87 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 168 or its variants containing up to approximately 3 amino acid substitutions; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 102 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 183 or its variants containing up to approximately 3 amino acid substitutions; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 105 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 186 or its variants containing up to approximately 3 amino acid substitutions; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 108 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 189 or its variants containing up to approximately 3 amino acid substitutions; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 115 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 196 or its variants containing up to approximately 3 amino acid substitutions; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 120 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 201 or its variants containing up to approximately 3 amino acid substitutions; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 88 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 169 or its variants containing up to approximately 3 amino acid substitutions; (8) CDR1 containing the amino acid sequence of SEQ ID NO: 8 or a variant thereof containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 89 or a variant thereof containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 170 or a variant thereof containing up to approximately 3 amino acid substitutions; or (9) CDR1 containing the amino acid sequence of SEQ ID NO: 9 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 90 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 171 or its variants containing up to approximately 3 amino acid substitutions. Includes a polypeptide containing one of the following.

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

[0034] In some embodiments, the sdAb portion is a camel sdAb produced in human or rhesus monkey DLL3.

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

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

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

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

[0039] In another embodiment, the disclosure relates to a DLL3-binding protein comprising a single-stranded variable fragment (scFv) moiety that specifically binds to DLL3, wherein the scFv moiety comprises a heavy-chain variable region (VH) and a light-chain variable region (VL), and the VH domain of the scFv moiety comprises CDR1 or a variant thereof, represented by SEQ ID NOs. 498 or 504, CDR1 or a variant thereof, containing up to about three amino acid substitutions; CDR2 or a variant thereof, represented by SEQ ID NOs. 499 or 505, CDR2 or a variant thereof, containing up to about three amino acid substitutions; and CD, represented by SEQ ID NOs. 500 or 506. The present invention provides a DLL3-binding protein comprising a variant of R3 or CDR3 containing up to approximately three amino acid substitutions, wherein the VL domain of the scFv portion comprises CDR1 or a variant of CDR1 containing up to approximately three amino acid substitutions, as shown in SEQ ID NO: 495 or 501, CDR2 or a variant of CDR2 containing up to approximately three amino acid substitutions, as shown in SEQ ID NO: 496 or 502, and CDR3 or a variant of CDR3 containing up to approximately three amino acid substitutions, as shown in SEQ ID NO: 497 or 503.

[0040] In some embodiments, the scFv portion includes a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH domain of the scFv portion includes 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 domain of the scFv portion includes 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 domain of the scFv portion includes 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 domain of the scFv portion includes CDR1 shown in SEQ ID NO: 501, CDR2 shown in SEQ ID NO: 502, and CDR3 shown in SEQ ID NO: 503.

[0041] In some embodiments, the VH domain of the scFv portion contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 508, and the VL domain of the scFv portion contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 507; or the VH domain of the scFv portion contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 510, and the VL domain of the scFv portion contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown 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, DLL3 is a human or a rhesus monkey DLL3.

[0044] In another aspect, the disclosure provides isolated nucleic acid molecules encoding anti-DLL3 CAR or DLL3-binding proteins as described above.

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

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

[0047] In some embodiments, the isolated nucleic acid molecule contains a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 511-514, which encodes the VL or VH domain of 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, PD-1 DNR includes an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 523.

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

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

[0053] In some embodiments, a polynucleotide sequence encoding PD-1 DNR, PD-1 CSR, or TGF-β DNR is ligated to a polynucleotide sequence encoding 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 PD-1 DNR, PD-1 CSR, or TGF-β DNR.

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

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

[0058] In another embodiment, the present disclosure provides modified immune cells containing the isolated nucleic acid molecules described above.

[0059] In some embodiments, the modified immune cells are selected from a group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, NKT cells, and Nature Killer cells.

[0060] In another aspect, the disclosure provides modified immune cells expressing the anti-DLL3 CAR described above.

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

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

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

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

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

[0066] In some embodiments, CAR and CSR, or CAR and DNR, are expressed simultaneously 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 modified immune cells are stimulated by cells that express CAR and PD-1 DNR, and also express DLL3 and PD-L1.

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

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

[0071] In some embodiments, the modified 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 nature killer cells.

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

[0073] In another aspect, the Disclosure provides a method for treating a DLL3-related disorder in a subject, comprising administering a therapeutically effective amount of such modified immune cells or a therapeutically effective amount of such pharmaceutical composition to the subject.

[0074] In another aspect, the disclosure provides the use of such anti-DLL3 CARs, isolated DLL3-binding proteins, expression vectors, or modified immune cells for the preparation of agents for treating DLL3-related disorders.

[0075] In another aspect, the present disclosure provides a drug for use in the treatment of DLL3-related disorders, comprising an anti-DLL3 CAR, a DLL3-binding protein, an expression vector, or modified immune cells as described above.

[0076] In some embodiments, DLL3-related disorders are cancers 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, cancer expresses DLL3 and PD-L1.

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

[0079] In some embodiments, DLL3-related disorders are associated with small cell lung cancer. [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-stranded variable fragment (scFv). [Invention 1002] The CAR of the present invention 1001 comprises a polypeptide in which the sdAb includes a CDR1 containing any one amino acid sequence of SEQ ID NOs. 1 to 81 or a variant thereof containing up to about three amino acid substitutions in CDR1, a CDR2 containing any one amino acid sequence of SEQ ID NOs. 82 to 162 or a variant thereof containing up to about three amino acid substitutions in CDR2, and a CDR3 containing any one amino acid sequence of SEQ ID NOs. 163 to 243 or a variant thereof containing up to about three amino acid substitutions in CDR3. [Invention 1003] The CAR of the present invention 1001 or 1002, wherein the sdAb comprises a polypeptide comprising CDR1 containing any one amino acid sequence of SEQ ID NOs. 1 to 81, CDR2 containing any one amino acid sequence of SEQ ID NOs. 82 to 162, and CDR3 containing any one amino acid sequence 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. [Invention 1004] The aforementioned sdAb is, (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 87 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 168 or its variants containing up to approximately 3 amino acid substitutions; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 102 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 183 or its variants containing up to approximately 3 amino acid substitutions; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 105 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 186 or its variants containing up to approximately 3 amino acid substitutions; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 108 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 189 or its variants containing up to approximately 3 amino acid substitutions; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 115 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 196 or its variants containing up to approximately 3 amino acid substitutions; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 120 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 201 or its variants containing up to approximately 3 amino acid substitutions; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 88 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 169 or its variants containing up to approximately 3 amino acid substitutions; (8) CDR1 containing the amino acid sequence of SEQ ID NO: 8 or a variant thereof containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 89 or a variant thereof containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 170 or a variant thereof containing up to approximately 3 amino acid substitutions; or (9) CDR1 containing the amino acid sequence of SEQ ID NO: 9 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 90 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 171 or its variants containing up to approximately 3 amino acid substitutions. A CAR according to any one of the present invention 1001 to 1003, comprising a polypeptide containing one of the following. [Invention 1005] The aforementioned sdAb is, (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6; CDR2 containing the amino acid sequence of SEQ ID NO: 87; and CDR3 containing the amino acid sequence of SEQ ID NO: 168; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21; CDR2 containing the amino acid sequence of SEQ ID NO: 102; and CDR3 containing the amino acid sequence of SEQ ID NO: 183; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24; CDR2 containing the amino acid sequence of SEQ ID NO: 105; and CDR3 containing the amino acid sequence of SEQ ID NO: 186; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27; CDR2 containing the amino acid sequence of SEQ ID NO: 108; and CDR3 containing the amino acid sequence of SEQ ID NO: 189; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34; CDR2 containing the amino acid sequence of SEQ ID NO: 115; and CDR3 containing the amino acid sequence of SEQ ID NO: 196; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39; CDR2 containing the amino acid sequence of SEQ ID NO: 120; and CDR3 containing the amino acid sequence of SEQ ID NO: 201; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7; CDR2 containing the amino acid sequence of SEQ ID NO: 88; and CDR3 containing the amino acid sequence of SEQ ID NO: 169; (8) CDR1 containing the amino acid sequence of SEQ ID NO: 8; CDR2 containing the amino acid sequence of SEQ ID NO: 89; and CDR3 containing the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 containing the amino acid sequence of SEQ ID NO: 9; CDR2 containing the amino acid sequence of SEQ ID NO: 90; and CDR3 containing the amino acid sequence of SEQ ID NO: 171. A CAR according to any one of the present invention 1001 to 1004, comprising a polypeptide containing one of the following. [Invention 1006] The CAR according to any one of the present invention 1001 to 1005, wherein the sdAb is a camel sdAb produced against human or rhesus monkey DLL3. [Invention 1007] The CAR according to any of the present invention 1001 to 1006, wherein the sdAb contains an amino acid sequence having at least about 95% sequence identity with any one of the amino acid sequences of SEQ ID NOs. 274 to 354. [Invention 1008] The CAR according to any of the present invention 1001 to 1007, wherein the sdAb has been humanized through CDR implantation. [Invention 1009] The humanized sdAb comprises an amino acid sequence having at least about 95% sequence identity with any one of the amino acid sequences of sequence numbers 355 to 367, according to any CAR of the present invention 1001 to 1008. [Invention 1010] The CAR according to any of the present invention 1001 to 1009, wherein the scFv comprises a heavy chain variable region (VH) and a light chain variable region (VL), and the VH of the scFv includes CDR1 represented by SEQ ID NO: 498 or 504 or a variant thereof containing up to about three amino acid substitutions in CDR1, CDR2 represented by SEQ ID NO: 499 or 505 or a variant thereof containing up to about three amino acid substitutions in CDR2, and CDR3 represented by SEQ ID NO: 500 or 506 or a variant thereof containing up to about three amino acid substitutions in CDR3, and the VL of the scFv includes CDR1 represented by SEQ ID NO: 495 or 501 or a variant thereof containing up to about three amino acid substitutions in CDR1, CDR2 represented by SEQ ID NO: 496 or 502 or a variant thereof containing up to about three amino acid substitutions in CDR2, and CDR3 represented by SEQ ID NO: 497 or 503 or a variant thereof containing up to about three amino acid substitutions in CDR3. [Invention 1011] A CAR according to any of the present inventions 1001 to 1010, wherein the VH of the scFv includes 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 includes CDR1 shown in SEQ ID NO: 495, CDR2 shown in SEQ ID NO: 496, and CDR3 shown in SEQ ID NO: 497; or a CAR according to any of the present inventions 1001 to 1010, wherein the VH of the scFv includes 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 includes CDR1 shown in SEQ ID NO: 501, CDR2 shown in SEQ ID NO: 502, and CDR3 shown in SEQ ID NO: 503. [Invention 1012] A CAR according to any of the present invention 1001 to 1011, wherein the VH of the scFv contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 508, and the VL of the scFv contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 507; or the VH of the scFv contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 510, and the VL of the scFv contains an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 509. [Invention 1013] The scFv is a CAR according to any of the inventions 1001 to 1012 obtained from a synthetic human Fab phage library. [Invention 1014] The CAR according to any of the present invention 1001 to 1013, wherein the DLL3 is a human or rhesus monkey DLL3. [Invention 1015] A CAR according to any one of the present invention 1001 to 1014, comprising a signal peptide, the DLL3 binding domain, a hinge domain, a transmembrane domain, and an intracellular signal transduction domain, from the N-terminus to the C-terminus. [Invention 1016] The CAR according to any of the present invention 1001 to 1015, wherein the intracellular signaling domain is derived from CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, or CD66d. [Invention 1017] The CAR according to any one of the present invention 1001 to 1016, wherein the intracellular signaling domain further comprises an intracellular costimulatory sequence. [Invention 1018] The CAR according to any of the present invention 1001 to 1017, wherein the intracellular co-stimulatory sequence is derived from a co-stimulatory 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, the ligand of CD83, and combinations thereof. [Invention 1019] A CAR according to any of the present invention 1001 to 1018, comprising an amino acid sequence having at least about 95% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 476-484, SEQ ID NOs: 485-494, or SEQ ID NOs: 515-516. [Invention 1020] The CAR according to any of the invention 1001 to 1019, wherein the DLL3 binding domain includes two sdAbs linked together. [Invention 1021] Each of the sdAb independently contains an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 356 or SEQ ID NO: 366, any CAR according to any of the present invention 1001 to 1020. [Invention 1022] A CAR according to any of the present invention 1001 to 1021, comprising an amino acid sequence having at least about 95% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs. 518 to 520. [Invention 1023] A CAR according to any of the present invention 1001 to 1022, comprising the amino acid sequence of SEQ ID NO: 520. [Invention 1024] A DLL3-binding protein comprising a single-domain antibody (sdAb) moiety that specifically binds to DLL3, wherein the sdAb moiety comprises a polypeptide comprising a CDR1 containing any one amino acid sequence of SEQ ID NOs. 1 to 81 or a variant thereof containing up to approximately three amino acid substitutions in the CDR1, a CDR2 containing any one amino acid sequence of SEQ ID NOs. 82 to 162 or a variant thereof containing up to approximately three amino acid substitutions in the CDR2, and a CDR3 containing any one amino acid sequence of SEQ ID NOs. 163 to 243 or a variant thereof containing up to approximately three amino acid substitutions in the CDR3. [Invention 1025] The DLL3-binding protein of the present invention 1024, wherein the sdAb portion comprises a polypeptide containing CDR1 containing any one amino acid sequence of SEQ ID NOs. 1 to 81, CDR2 containing any one amino acid sequence of SEQ ID NOs. 82 to 162, and CDR3 containing any one amino acid sequence of SEQ ID NOs. 163 to 243, or a variant of the polypeptide containing up to about three amino acid substitutions in CDR1, CDR2, and CDR3. [Invention 1026] The aforementioned sdAb portion, (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 87 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 168 or its variants containing up to approximately 3 amino acid substitutions; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 102 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 183 or its variants containing up to approximately 3 amino acid substitutions; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 105 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 186 or its variants containing up to approximately 3 amino acid substitutions; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 108 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 189 or its variants containing up to approximately 3 amino acid substitutions; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 115 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 196 or its variants containing up to approximately 3 amino acid substitutions; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 120 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 201 or its variants containing up to approximately 3 amino acid substitutions; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 88 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 169 or its variants containing up to approximately 3 amino acid substitutions; (8) CDR1 containing the amino acid sequence of SEQ ID NO: 8 or a variant thereof containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 89 or a variant thereof containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 170 or a variant thereof containing up to approximately 3 amino acid substitutions; or (9) CDR1 containing the amino acid sequence of SEQ ID NO: 9 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 90 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 171 or its variants containing up to approximately 3 amino acid substitutions. A DLL3-binding protein of the present invention 1024 or 1025, comprising a polypeptide containing one of the following. [Invention 1027] The aforementioned sdAb portion, (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6; CDR2 containing the amino acid sequence of SEQ ID NO: 87; and CDR3 containing the amino acid sequence of SEQ ID NO: 168; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21; CDR2 containing the amino acid sequence of SEQ ID NO: 102; and CDR3 containing the amino acid sequence of SEQ ID NO: 183; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24; CDR2 containing the amino acid sequence of SEQ ID NO: 105; and CDR3 containing the amino acid sequence of SEQ ID NO: 186; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27; CDR2 containing the amino acid sequence of SEQ ID NO: 108; and CDR3 containing the amino acid sequence of SEQ ID NO: 189; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34; CDR2 containing the amino acid sequence of SEQ ID NO: 115; and CDR3 containing the amino acid sequence of SEQ ID NO: 196; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39; CDR2 containing the amino acid sequence of SEQ ID NO: 120; and CDR3 containing the amino acid sequence of SEQ ID NO: 201; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7; CDR2 containing the amino acid sequence of SEQ ID NO: 88; and CDR3 containing the amino acid sequence of SEQ ID NO: 169; (8) CDR1 containing the amino acid sequence of SEQ ID NO: 8; CDR2 containing the amino acid sequence of SEQ ID NO: 89; and CDR3 containing the amino acid sequence of SEQ ID NO: 170; or (9) CDR1 containing the amino acid sequence of SEQ ID NO: 9; CDR2 containing the amino acid sequence of SEQ ID NO: 90; and CDR3 containing the amino acid sequence of SEQ ID NO: 171. A DLL3-binding protein according to any one of the invention 1024 to 1026, comprising a polypeptide containing one of the following. [Invention 1028] A DLL3-binding protein according to any one of the present invention 1024 to 1027, wherein the sdAb portion is a camel sdAb produced against human or rhesus monkey DLL3. [Invention 1029] A DLL3-binding protein according to any of the invention 1024 to 1028, wherein the sdAb portion contains an amino acid sequence having at least about 95% sequence identity with any one of the amino acid sequences of sequence numbers 274 to 354. [Invention 1030] A DLL3-binding protein according to any of the inventions 1024 to 1029, wherein the sdAb portion is humanized through CDR transplantation. [Invention 1031] A DLL3-binding protein according to any one of the invention 1024 to 1030, wherein the humanized sdAb contains an amino acid sequence having at least about 95% sequence identity with any one of the amino acid sequences of SEQ ID NOs. 355 to 367. [Invention 1032] The DLL3-binding protein according to any of the invention items 1024 to 1031, wherein the DLL3 is human or rhesus monkey DLL3. [Invention 1033] A DLL3-binding protein comprising a single-stranded 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), and the VH portion of the scFv portion is CDR1 represented by SEQ ID NO: 498 or 504 or a variant thereof containing up to approximately 3 amino acid substitutions in CDR1, CDR2 represented by SEQ ID NO: 499 or 505 or a variant thereof containing up to approximately 3 amino acid substitutions in CDR2, and CDR3 represented by SEQ ID NO: 500 or 506 or A DLL3-binding protein comprising a variant of CDR3 containing up to approximately three amino acid substitutions, wherein the VL of the scFv portion comprises CDR1 represented by SEQ ID NO: 495 or 501 or a variant of CDR1 containing up to approximately three amino acid substitutions, CDR2 represented by SEQ ID NO: 496 or 502 or a variant of CDR2 containing up to approximately three amino acid substitutions, and CDR3 represented by SEQ ID NO: 497 or 503 or a variant of CDR3 containing up to approximately three amino acid substitutions. [Invention 1034] The DLL3-binding protein of the present invention 1033, wherein the VH of the scFv portion includes 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 portion includes CDR1 shown in SEQ ID NO: 495, CDR2 shown in SEQ ID NO: 496, and CDR3 shown in SEQ ID NO: 497; or the DLL3-binding protein of the present invention 1033, wherein the VH of the scFv portion includes 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 portion includes CDR1 shown in SEQ ID NO: 501, CDR2 shown in SEQ ID NO: 502, and CDR3 shown in SEQ ID NO: 503. [Invention 1035] The DLL3-binding protein of the present invention 1033 or 1034, wherein the VH of the scFv portion includes an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 508, and the VL of the scFv portion includes an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 507; or the DLL3-binding protein of the present invention 1033 or 1034, wherein the VH of the scFv portion includes an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 510, and the VL of the scFv portion includes an amino acid sequence having at least about 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 509. [Invention 1036] The scFv portion is a DLL3-binding protein of any of the present invention 1033 to 1035 obtained from a synthetic human Fab phage library. [Invention 1037] The DLL3-binding protein according to any one of the present invention 1033 to 1036, wherein the DLL3 is human or rhesus monkey DLL3. [Invention 1038] An isolated nucleic acid molecule encoding any CAR according to Invention 1001-1023 or any DLL3-binding protein according to Invention 1024-1032 or Invention 1033-1037. [Invention 1039] An isolated nucleic acid molecule of the present invention 1038, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 368-448, which encodes a camel single-domain antibody (sdAb). [Invention 1040] An isolated nucleic acid molecule of the present invention 1038 or 1039, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs. 449 to 461, which encodes humanized camel sdAb. [Invention 1041] An isolated nucleic acid molecule according to any of the present invention 1038 to 1040, comprising a polynucleotide sequence selected from the group consisting of SEQ ID NOs. 511 to 514, which encodes the VL or VH domain of human scFv. [Invention 1042] An isolated nucleic acid molecule according to any of the present invention 1038 to 1041, further comprising a polynucleotide sequence encoding a chimeric switch receptor (CSR) or a dominant-negative receptor (DNR). [Invention 1043] An isolated nucleic acid molecule according to any of the present invention 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). [Invention 1044] An isolated nucleic acid molecule according to any of the present invention 1038 to 1043, wherein the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 523; and the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 524. [Invention 1045] An isolated nucleic acid molecule according to any of the present invention 1038 to 1044, wherein the TGF-β DNR contains an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 529. [Invention 1046] An isolated nucleic acid molecule according to any of the present invention 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. [Invention 1047] An isolated nucleic acid molecule according to any of the present invention 1038 to 1046, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide. [Invention 1048] An isolated nucleic acid molecule according to any of invention 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. [Invention 1049] An isolated nucleic acid molecule according to any of the Invention 1038 to 1048, which encodes a peptide having at least about 95% sequence identity with SEQ ID NO: 521 or 522, or a peptide having at least about 95% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NOs: 525 to 528. [Invention 1050] An expression vector comprising any isolated nucleic acid molecule according to invention 1038 to 1049. [Invention 1051] Modified immune cells comprising any isolated nucleic acid molecule according to Invention 1038-1049 or an expression vector according to Invention 1050. [Invention 1052] A modified immune cell according to 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. [Invention 1053] Modified immune cells expressing any of the CARs described in invention 1001 to 1023. [Invention 1054] A modified immune cell according to Invention 1053, which also expresses CSR or DNR. [Invention 1055] A modified immune cell according to invention 1053 or 1054, wherein the CSR is PD-1 CSR and the DNR is PD-1 DNR or TGF-β DNR. [Invention 1056] A modified immune cell according to any of invention 1053 to 1055, wherein the PD-1 DNR comprises an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 523; and the PD-1 CSR comprises an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 524. [Invention 1057] A modified immune cell according to any of the present invention 1053 to 1056, wherein the TGF-β DNR contains an amino acid sequence having at least about 95% sequence identity with SEQ ID NO: 529. [Invention 1058] A modified immune cell according to any of the present invention 1053 to 1057, wherein the CAR and the CSR, or the CAR and the DNR, are simultaneously expressed via a 2A self-cleaving peptide. [Invention 1059] A modified immune cell according to any one of the invention items 1053 to 1058, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide. [Invention 1060] A modified immune cell according to any of the inventions 1053 to 1059, expressing the aforementioned CAR and the aforementioned PD-1 CSR. [Invention 1061] A modified immune cell according to any of items 1053 to 1060 of the present invention, which expresses the aforementioned CAR and the aforementioned TGF-β DNR, and is stimulated by a cell expressing DLL3 in the presence of TGF-β. [Invention 1062] A modified immune cell according to any of the invention 1053 to 1061, 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. [Invention 1063] A pharmaceutical composition comprising any CAR according to Invention 1001 to 1023, any DLL3-binding protein according to Invention 1024 to 1032 or Invention 1033 to 1037, an expression vector according to Invention 1050, or a modified immune cell according to any 1051 to 1062, and a physiologically acceptable excipient. [Invention 1064] A method for treating DLL3-related disorders in a subject, comprising administering to the subject a therapeutically effective amount of any modified immune cells according to Invention 1051 to 1062, or a therapeutically effective amount of a pharmaceutical composition according to Invention 1063. [Invention 1065] The method of the present invention 1064, wherein the 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. [Invention 1066] The method of the present invention 1065, wherein the DLL3-related disorder is small cell lung cancer. [Invention 1067] The method according to any one of the present invention 1064 to 1066, wherein the cancer expresses DLL3 and PD-L1. [Invention 1068] The method according to any one of the present invention 1064 to 1067, wherein the cancer has a higher TGF-β expression level compared to the corresponding normal tissue. [Invention 1069] Use of any CAR of Invention 1001-1023, any DLL3-binding protein of Invention 1024-1032 or Invention 1033-1037, expression vector of Invention 1050, or modified immune cells of Invention 1051-1062 for the preparation of agents for treating DLL3-related disorders. [Invention 1070] A drug for use in the treatment of DLL3-related disorders, comprising any CAR of Invention 1001 to 1023, any DLL3-binding protein of Invention 1024 to 1032 or Invention 1033 to 1037, an expression vector of Invention 1050, or modified immune cells of any of Invention 1051 to 1062. [Brief explanation of the drawing]

[0080] [Figure 1] Figure 1 shows a schematic diagram of a CAR structure based on VHH. The sequences available in the structure are listed in sequence numbers 462-472, 474, and 475. [Figure 2] Figure 2 shows the results of in vitro cytotoxicity assays against the small cell lung cancer cell line SHP-77 using T cells expressing exemplary monospecific CARs, including various camel anti-DLL3 sdAbs, in 2:1 or 5:1 E:T ratios. The results in CARs are presented in the order shown in the legend on the right. [Figure 3]Figure 3 shows the cytokine release levels of T cells expressing exemplary monospecific CARs, including various camel anti-DLL3 sdAbs, after co-incubation with the DLL3-expressing tumor cell line SHP-77. IFN-γ release levels and TNF-α release levels (E:T ratio 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 the order shown in the legend on the right. [Figure 4] Figure 4 shows the expanded proliferation rate of T cells expressing exemplary monospecific CARs, including various camel 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 CARs containing camel anti-DLL3 sdAb in the SHP-77 tumor model. In this model, 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, including various humanized camel anti-DLL3 sdAbs, against small cell lung cancer cell lines SHP-77 (Figure 6A, Figure 6B) and NCI-H82 (Figure 6C, Figure 6D). [Figure 6B] See the explanation in Figure 6A. [Figure 6C] See the explanation in Figure 6A. [Figure 6D] See the explanation in Figure 6A. [Figure 7] Figure 7 shows the cytokine release levels of T cells expressing exemplary monospecific CARs, including various humanized camel anti-DLL3 sdAbs, after stimulation with SHP-77. IFN-γ release levels and TNF-α release levels (E:T ratio 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 the order shown in the legend on the right. [Figure 8] Figure 8 shows the expanded proliferation rate of T cells expressing exemplary monospecific CARs, including various humanized camel 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 humanized camel anti-DLL3 sdAb in a 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 within each group are shown in Figures 9B-9J. [Figure 9B] See the explanation in Figure 9A. [Figure 9C] See the explanation in Figure 9A. [Figure 9D] See the explanation in Figure 9A. [Figure 9E] See the explanation in Figure 9A. [Figure 9F] See the explanation in Figure 9A. [Figure 9G] See the explanation in Figure 9A. [Figure 9H] See the explanation in Figure 9A. [Figure 9I] See the explanation in Figure 9A. [Figure 9J] See the explanation in Figure 9A. [Figure 10A] Figure 10 shows a schematic diagram of a tandem CAR (Figure 10A) and schematic diagrams of an armed CAR structure (Figures 10B and 10C). [Figure 10B] Refer to the explanation in Figure 10A. [Figure 10C] Refer to the explanation in Figure 10A. [Figure 11A] Figure 11 shows the results of a comparison of the in vitro functional activity of tandem CAR-T cells and monospecific CAR-T cells based on measurements of short-term cytotoxicity (Figures 11A-E, 11V), cytokine release (Figures 11F-K, 11W-X), and long-term stimulation assays (Figures 11L-U, 11Y-Z). [Figure 11B] Refer to the explanation in Figure 11A. [Figure 11C] Refer to the explanation in Figure 11A. [Figure 11D] Refer to the explanation in Figure 11A. [Figure 11E] Refer to the explanation in Figure 11A. [Figure 11F] Refer to the explanation in Figure 11A. [Figure 11G] Refer to the explanation in Figure 11A. [Figure 11H] Refer to the explanation in Figure 11A. [Figure 11I] Refer to the explanation in Figure 11A. [Figure 11J] Refer to the explanation in Figure 11A. [Figure 11K] Refer to the explanation in Figure 11A. [Figure 11L] Refer to the explanation in Figure 11A. [Figure 11M] Refer to the explanation in Figure 11A. [Figure 11N] Refer to the explanation in Figure 11A. [Figure 11O] Refer to the explanation in Figure 11A. [Figure 11P] Refer to the explanation in Figure 11A. [Figure 11Q] Refer to the explanation in Figure 11A. [Figure 11R] Refer to the explanation in Figure 11A. [Figure 11S] Refer to the explanation in Figure 11A. [Figure 11T] Refer to the explanation in Figure 11A. [Figure 11U] Refer to the explanation in Figure 11A. [Figure 11V] Refer to the explanation in Figure 11A. [Figure 11W] Refer to the explanation in Figure 11A. [Figure 11X] Refer to the explanation in Figure 11A. [Figure 11Y] Refer to the explanation in Figure 11A. [Figure 11Z] Refer to the explanation in Figure 11A. [Figure 12A] Figure 12 shows the in vitro functional comparison results of CAR-T cells and T3 cells armed with PD-1 DNR or PD-1 CSR, targeting SHP-77 cells (Figure 12A, Figure 12B) and SHP-77 / PD-L1 (Figure 12C, Figure 12D), respectively. [Figure 12B] Refer to the explanation in Figure 12A. [Figure 12C] Refer to the explanation in Figure 12A. [Figure 12D] Refer to the explanation in 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 positivity rates for each CAR-T cell type, CAR and TGF-β-DNR. In vitro antitumor efficacy was evaluated in short-term stimulation assays (Figures 13C, 13D) and long-term stimulation assays (Figures 13E-G). Figure 13H shows the in vivo antitumor efficacy. Figure 13I shows the pharmacokinetics of CAR-T cells in peripheral blood of a SHP77 xenograft model after treatment. [Figure 13B] Refer to the explanation in Figure 13A. [Figure 13C] Refer to the explanation in Figure 13A. [Figure 13D] Refer to the explanation in Figure 13A. [Figure 13E] Refer to the explanation in Figure 13A. [Figure 13F] Refer to the explanation in Figure 13A. [Figure 13G] Refer to the explanation in Figure 13A. [Figure 13H] Refer to the explanation in Figure 13A. [Figure 13I] Refer to the explanation in Figure 13A. [Modes for carrying out the invention]

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

[0082] The articles “a” and “an” are used herein to refer to one or more (i.e., at least one) grammatical objects of the articles. For example, “element” means one or more elements.

[0083] The term “binding protein,” as used herein, refers to a molecule or part of a molecule that binds to a target molecule (e.g., DLL3). In some embodiments, the binding protein includes an antibody. In some embodiments, the binding protein includes an antigen-binding fragment of an antibody. In some embodiments, the binding protein may further include a low molecular weight component, such as a small molecule drug or toxin. The binding protein may also be an antibody or its antigen-binding fragment. In some embodiments, the binding protein includes the ligand-binding domain of a receptor. In some embodiments, the binding protein includes 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 includes a single-domain antibody (sdAb) or a single-strand variable fragment (scFv). In some embodiments, the binding protein may be an sdAb or scFv. The DLL3-binding protein may be a DLL3-binding domain. In some embodiments, the DLL3-binding protein includes 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-stranded variable fragment (scFv) that binds to DLL3. In some embodiments, the DLL3-binding protein may be either an sdAb or a 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 retain the essential epitope-binding characteristics of the Ig molecule. In a full-length antibody, 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 hypervariable regions called complementarity-determining regions (CDRs), which are scattered within more conserved regions called framework regions (FRs). Each VH and VL consists of three CDRs and four FRs arranged in the order FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4 from the amino terminus to the carboxyl terminus. Immunoglobulin molecules can be 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 broader sense, the term “antibody” further refers to scFv or sdAb that do not originate from immunoglobulin molecules having four polypeptide chains.

[0085] Antibody fragments are parts of antibodies such as F(ab')2, Fab, Fv, scFv, and sdAb. Functional fragments of full-length antibodies retain the target specificity of the full-length antibody. Therefore, recombinant functional antibody fragments, such as scFv (single-stranded variable-chain fragments), are used to develop therapeutics as alternatives to mAb-based therapies. The scFv fragment (approximately 25 kDa) consists of two variable domains, VH and VL. In nature, the VH and VL domains tend to associate non-covalently via hydrophobic interactions and dissociate. However, scFv can be created by modifying stable fragments by linking the domains with hydrophilic flexible linkers.

[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 of a type found in camelid mammals that naturally lacks a light chain. Such single-domain antibodies are V H Also referred to as H or "nanobody". The amino acid sequence and structure of a single-domain antibody can be considered to consist of four framework regions (FR1, FR2, FR3, and FR4) and three complementarity-determining regions (CDR1, CDR2, and CDR3). Therefore, a single-domain antibody can be defined as an amino acid sequence having a general structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4, similar to the variable domain VH or VL. The use of sdAbs as a single antigen-binding protein or as an antigen-binding domain in a larger protein or polypeptide offers several significant advantages over the use of conventional antibodies or antibody fragments (e.g., scFv). The advantages of sdAbs include the requirement that only a single domain binds to the antigen with high affinity and selectivity; that sdAbs are highly stable to denaturing factors or conditions including heat, pH, and proteases; and that sdAbs can access targets and epitopes that conventional antibodies cannot access. Typically, sdAb is produced in camels such as llamas, but it can also be synthesized using techniques well known in the art.

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

[0088] As used herein, the term “heavy chain monoantibody” 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. First identified in the nematode C. elegans and Drosophila, and subsequently shown to be evolutionarily conserved from invertebrates to 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 serate and delta (Wharton et al., 1985; Rebay et al., 1991). In humans, four known Notch receptors and five DSL (delta-serate LAG2) ligands are recognized—two homologs of serate known as jagged 1 and jagged 2, and three homologs of delta, referred to as delta-like ligands or DLL1, DLL3, and DLL4. In humans, the DLL3 gene is located on chromosome 19q13 and consists of eight exons totaling 9.5kb. Alternative splicing within the final exon results in two protein isoforms. Both share 100% overall identity through their extracellular and transmembrane domains, differing only in that the longer isoform has an extended cytoplasmic end.

[0090] As used herein, the terms “specifically bind” or “specifically bound” or any synonyms 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 an assay by a standard in vitro assay. For example, binding, as used herein, is measured by the ability of the anti-DLL3 polypeptide of the present invention to recognize a DLL3 molecule on a cell surface using a well-described antigen-antibody binding assay, flow cytometry, and other assays known to those skilled in the art.

[0091] As used herein, the term “expression vector” is a nucleic acid construct or sequence constructed recombinantly or synthetically using specific nucleic acid factors that enable the transcription and / or expression of another nucleic acid in a host cell. An expression vector may be part of a plasmid, virus, or nucleic acid fragment. In one example, an expression vector is a DNA vector, e.g., a plasmid, comprising at least one promoter sequence and at least one terminator sequence (e.g., a polyadenylation sequence), and optionally an origin of replication (ori) sequence, and optionally a selectable or selectable marker sequence. Optionally, an expression vector may further comprise at least one nucleotide coding sequence of interest encoding at least one polypeptide, wherein at least one promoter sequence is operably ligated with at least one coding sequence. The term “expression” includes, but is not limited to, any steps involved in the production of a polypeptide, including transcription, post-transcriptional modification, translation, post-translational modification, and / or secretion.

[0092] The term “isolated” typically refers to material that has been substantially or essentially freed from components present in its natural state. The material may 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 purified from its adjacent sequences in its naturally occurring state, such as a DNA fragment typically removed from adjacent sequences. Alternatively, “isolated antibody” or “isolated polypeptide,” as used herein, refers to the in vitro isolation and / or purification of an antibody or polypeptide molecule from its natural cellular environment and its association with other cellular components.

[0093] The "humanized" form of a non-human (e.g., camel) antibody is a chimeric antibody having a minimal sequence derived from a non-human immunoglobulin. In some embodiments, the humanized antibody is a human immunoglobulin (recipient antibody) in which residues from the recipient's hypervariable region (HVR) are replaced with residues from the HVR of a non-human species (donor antibody), such as mouse, rat, rabbit, or non-human primate, having the desired specificity, affinity, and / or capabilities. In some cases, framework ("FR") residues of the human immunoglobulin are replaced with corresponding non-human residues. Furthermore, the humanized antibody may contain residues not found in the recipient antibody or the donor antibody. These modifications may be made to further refine antibody performance, such as binding affinity. Generally, humanized antibodies will contain at least one, typically two, variable domains substantially, all or substantially all of the hypervariable loops corresponding to those of a non-human immunoglobulin sequence, and all or substantially all of the FR region being equivalent to that of a human immunoglobulin sequence, although the FR region may contain one or more FR residue substitutions that improve antibody performance, such as binding affinity, isomerization, and immunogenicity. The number of these amino acid substitutions in the FR is typically six or less in the H chain and three or less in the L chain. Humanized antibodies will also optionally contain at least a portion of the immunoglobulin constant region (Fc), typically that of human immunoglobulin. For further details, see, for example, 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). Furthermore, see, for example, 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 No. 6,982,321 and U.S. Patent No. 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 amino acid residues in a particular peptide or polypeptide sequence after aligning the sequences, introducing gaps as necessary, and achieving maximum percentage sequence identity without considering any conservative substitutions as part of sequence identity. Alignment intended to determine the percentage of amino acid sequence identity can be achieved in various ways within the scope 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 algorithm required to achieve maximum alignment over the full length of the sequences being compared.

[0095] Dissociation constant (K D or K d K) is used as an indicator of the affinity of an antibody to an antigen. For example, it can be easily analyzed by using the scatchard method, which uses antibodies labeled with various marker agents, and by using the OTC measurement kit BiacoreX (manufactured by Amersham Biosciences) or a similar kit, following the user manual and experimental procedure instructions provided with the kit. K that can be derived using these methods D The value is expressed in moles (M).

[0096] When used herein, “chimeric switch receptor (CSR)” refers to a receptor constructed to reverse the outcome of its original signaling pathway in order to confer desired activity to immune cells (e.g., CAR T cells), such as the ability to overcome an immunosuppressive tumor microenvironment and to have greater in vivo persistence. In some embodiments, the CSR can further stimulate CAR T cells by utilizing inhibitory molecules expressed by cancer cells. In non-limiting examples, CAR T cells may be modified 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 CAR T cells are administered to a subject with cancer expressing DLL3 and programmed cell death ligand 1 (PD-L1), the expressed CAR may bind to DLL3, and the expressed switch receptor may bind to PD-L1. The properties of the PD-1 / CD28 chimeric switch receptor fusion protein are such that it blocks normal PD1 / PD-L1-mediated T cell repression and instead promotes signaling through the CD28 domain, leading to stimulation of CAR T cells. Therefore, by exchanging the transmembrane and intracellular domains of PD-1 with those of CD28, PD-L1 is converted into a costimulatory ligand for CAR T cells. This induces enhanced toxicity against cancer cells expressing PD-L1. In other embodiments, CSRs can also be used to inhibit the effects of CAR T cells on unintended target cells.

[0097] When used herein, “dominant-negative receptor (DNR)” refers to a receptor that can bind to its ligand but does not induce a signaling cascade within the cell. DNRs typically have an intact ligand-binding domain but lack an intracellular enzymatic domain. They may be mutant forms of the full-length receptor or cleaved forms of the receptor. After CAR T cell immunotherapy, some cancers, particularly solid tumors, 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 tumors, 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 to potently suppress the immune system. Therefore, in some embodiments, the inventors improve the antitumor performance of the CAR T cells disclosed herein by using a cleaved version of the TGF-β receptor TGF-BRII as a TGF-β DNR. In some embodiments, CAR and TGF-β DNR are expressed simultaneously on the surface of T cells by the use of a 2A self-cleaving peptide. In some embodiments, CAR and TGF-β DNR are expressed separately on the surface of T cells by the use of two expression vectors. We have found that when TGF-β DNR is introduced into anti-DLL3 CAR T cells disclosed herein, it can enhance the cytotoxicity of CAR T cells against certain DLL3-positive cancer cells, such as SCLC cells. Similarly, in some embodiments, we improve the antitumor performance of CAR T cells disclosed herein by using a cleaved version of the PD-1 receptor as PD-1 DNR.

[0098] As used herein, “treatment” or “to treat” includes any beneficial or desirable 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, such as cancer, autoimmune disease, or immune disorder. Treatment may optionally include delaying the progression of the disease or condition. “Treatment” does not necessarily imply the complete eradication or cure of the disease or condition or its associated symptoms.

[0099] Some aspects of the present invention relate to DLL3-binding proteins having binding specificity to human or rhesus monkey DLL3 proteins.

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

[0101] In some embodiments, the sdAb comprises a polypeptide comprising CDR1 containing any one amino acid sequence of SEQ ID NOs: 1-81; CDR2 containing any one amino acid sequence of SEQ ID NOs: 82-162; and CDR3 containing any one amino acid sequence of SEQ ID NOs: 163-243; or a variant of a polypeptide containing up to about three amino acid substitutions in the CDR region. 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 monkey DLL3 protein. In some embodiments, the sdAb comprises a CDR set (i.e., a combination of CDR1, CDR2, and CDR3) as listed in each column of Table 1.

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

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

[0104] In some embodiments, 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 any one of the amino acid sequences of SEQ ID NOs. 274–354. In other embodiments, sdAb comprises a humanized amino acid sequence having at least about 95% (e.g., about 96%, about 97%, about 98%, about 99%, or about 100%) sequence identity to any one of the amino acid sequences of SEQ ID NOs. 355–367. In some embodiments, sdAb comprises any one of the amino acid sequences 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 transplantation (see, e.g., U.S. Patents No. 5,225,539, 5,530,101 and 5,585,089), veneering or resurfacing (see, e.g., European Patent No. 592,106 and 519,596) and chain shuffling (see, e.g., U.S. Patent No. 5,565,332). Generally, during humanization, CDR residues of receptor antibodies (e.g., human antibodies) are substituted with CDR residues of donor antibodies (e.g., rodent antibodies) to maintain antigen-binding specificity while minimizing in vivo immunogenicity. In many cases, the substitution of framework residues within the framework region with corresponding residues from the donor antibody can also modify, for example, improve, antigen-binding properties. These framework substitutions, such as conservative substitutions, are identified by methods well known in the art, such as CDRs for identifying framework residues important for antigen binding, modeling of framework residue interactions, and sequence comparisons for identifying abnormal framework residues at specific locations (see, for example, Queen et al., U.S. Patent No. 5,585,089; and Riechmann et al., 1988, Nature, 332:323).

[0105] In some cases, sdAbs can be fused with human IgG hinge and Fc fragments to form heavy chain antibodies (HCAbs). In some cases, sdAbs can be fused with another sdAb or scFv that is specific to an antigen other than DLL3 to form bispecific antibodies. In some cases, sdAbs can be fused with two or more sdAbs or scFv that are specific to an antigen other than DLL3 to form polyspecific antibodies. In other cases, sdAbs can be chemically modified to carry drug molecules. Therefore, anti-DLL3 sdAbs can be used in vivo to deliver drug molecules to DLL3-expressing cells.

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

[0107] In some cases, scFv can fuse with another sdAb or scFv that is specific to an antigen other than DLL3, thus forming a bispecific antibody. In some cases, scFv can fuse with two or more sdAbs or scFv that are specific to an antigen other than DLL3, thus forming a polyspecific antibody. In other cases, scFv can be chemically modified to carry a drug molecule. Therefore, anti-DLL3 scFv can be used in vivo to deliver drug molecules to DLL3-expressing cells.

[0108] Some aspects of the present invention relate to CAR or CAR-T cells (anti-DLL3 CAR or anti-DLL3 CAR-T cells) containing a DLL3 binding domain.

[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 contains a target-specific binding factor (also referred to as an antigen-recognition domain or antigen-binding domain). The intracellular domain, or otherwise the cytoplasmic domain, often contains one or more costimulatory signaling domains and a CD3ζ chain moiety. The costimulatory signaling domain refers to a portion of the CAR that includes the intracellular domain of the costimulatory molecule.

[0110] Antigen recognition or targeting by CAR molecules most commonly involves the use of antibodies or antibody fragments. 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 originate from either a natural or synthetic source. If the source is natural, the domain may originate from any membrane-bound or transmembrane protein. The transmembrane region of a particular use in the present invention may originate, for example, from the alpha, beta, or zeta chain of a T cell receptor, or the CD8α chain (i.e., it may include at least its transmembrane region).

[0112] The intracellular signaling domain of the CAR of the present invention is involved in the activation of 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 may be cytolytic activity or helper activity, including cytokine secretion. Therefore, the term “intracellular domain” refers to the portion of a protein that transmits effector function signals and guides the cell to perform its specialized function. Usually, the entire cytoplasmic domain can be used, but in many cases, it is not necessary to use the entire chain. Only cleavage portions of the cytoplasmic domain are used, and such cleavage portions can be used in place of the intact chain as long as they transmit effector function signals. Therefore, the term intracellular domain means that it includes any cleavage portion of the cytoplasmic domain sufficient to transmit effector function signals. Preferred examples of cytoplasmic domains intended for use in the CAR of the present invention include cytoplasmic sequences of T cell receptors (TCRs) and coreceptors that act in conjunction with the initiation of signaling after antigen receptor association, 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 CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, or CD66d.

[0113] In many cases, the signal generated by the TCR alone is insufficient for the complete activation of T cells. Therefore, secondary or costimulatory signals are used. Thus, it can be said that T cell activation is mediated by two distinct 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). Costimulatory signaling sequences refer to a portion of the CAR containing the intracellular domain of a costimulatory molecule. Costimulatory molecules are cell surface molecules other than antigen receptors or their ligands that are required for the efficient response of lymphocytes to an antigen. 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 domain and the transmembrane domain of a CAR generally refers to any oligonucleotide 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 to 100 amino acids, and most preferably 2 to 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 include a signal peptide sequence linked to the N-terminus of the CAR. Signal peptide sequences are present at the N-terminus of many secreted and membrane proteins and are typically 15 to 30 amino acids long. 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 includes a DLL3-binding domain, which includes or is derived from a single-domain antibody (sdAb) or a single-stranded variable fragment (scFv).

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

[0118] In some embodiments, the CAR includes a DLL3 binding domain, which is as follows: (1) CDR1 containing the amino acid sequence of SEQ ID NO: 6 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 87 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 168 or its variants containing up to approximately 3 amino acid substitutions; (2) CDR1 containing the amino acid sequence of SEQ ID NO: 21 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 102 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 183 or its variants containing up to approximately 3 amino acid substitutions; (3) CDR1 containing the amino acid sequence of SEQ ID NO: 24 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 105 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 186 or its variants containing up to approximately 3 amino acid substitutions; (4) CDR1 containing the amino acid sequence of SEQ ID NO: 27 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 108 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 189 or its variants containing up to approximately 3 amino acid substitutions; (5) CDR1 containing the amino acid sequence of SEQ ID NO: 34 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 115 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 196 or its variants containing up to approximately 3 amino acid substitutions; (6) CDR1 containing the amino acid sequence of SEQ ID NO: 39 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 120 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 201 or its variants containing up to approximately 3 amino acid substitutions; (7) CDR1 containing the amino acid sequence of SEQ ID NO: 7 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 88 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 169 or its variants containing up to approximately 3 amino acid substitutions; (8) CDR1 containing the amino acid sequence of SEQ ID NO: 8 or a variant thereof containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 89 or a variant thereof containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 170 or a variant thereof containing up to approximately 3 amino acid substitutions; or (9) CDR1 containing the amino acid sequence of SEQ ID NO: 9 or its variants containing up to approximately 3 amino acid substitutions; CDR2 containing the amino acid sequence of SEQ ID NO: 90 or its variants containing up to approximately 3 amino acid substitutions; and CDR3 containing the amino acid sequence of SEQ ID NO: 171 or its variants containing up to approximately 3 amino acid substitutions. Includes sdAb which contains one of the following.

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

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

[0121] In some embodiments, the CAR includes a DLL3 binding domain, the DLL3 binding domain includes or is derived from a single-stranded variable fragment (scFv), the VH domain of the scFv includes 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 domain of the scFv includes 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 domain of the scFv includes 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 domain of the scFv includes CDR1 shown in SEQ ID NO: 501, CDR2 shown in SEQ ID NO: 502, and CDR3 shown in SEQ ID NO: 503. In some embodiments, the VH domain of scFv comprises the amino acid sequence shown in SEQ ID NO: 508, and the VL domain of scFv comprises the amino acid sequence shown in SEQ ID NO: 507; or the VH domain of scFv comprises the amino acid sequence shown in SEQ ID NO: 510, and the VL domain of scFv comprises the amino acid sequence shown in SEQ ID NO: 509.

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

[0123] In some embodiments, the CAR of the present 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 any one of the amino acid sequences of SEQ ID NOs. 515-516. In some embodiments, the CAR of the present invention comprises a human scFv provided herein as a DLL3 binding domain and comprises an amino acid sequence selected from the group consisting of SEQ ID NOs. 515-516.

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

[0125] Some aspects of the present invention relate to isolated nucleic acid molecules encoding sdAb, scFv, or CAR. In some embodiments, the nucleic acid molecule encodes camel sdAb and includes a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 368 to 448. In some embodiments, the nucleic acid molecule encodes humanized sdAb and includes a polynucleotide sequence selected from the group consisting of SEQ ID NOs: 449 to 461. In some embodiments, the nucleic acid molecule encodes the VH and VL domains of scFv, the sequence encoding the VH domain includes the polynucleotide sequence of SEQ ID NOs: 512 or 514, and the sequence encoding the VL domain includes the polynucleotide sequence of SEQ ID NOs: 511 or 513.

[0126] Several aspects of the present application relate to modified immune cells comprising any one of the CARs provided above, or 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 the isolated nucleic acid molecule of the present invention. Genetically modifying cells with an expression vector for expressing a polypeptide encoded by a portion of a nucleic acid molecule is a well-known genetic technique in the art.

[0127] Some aspects of the present invention relate to the use of the DLL3-binding protein, anti-DLL3 CAR, 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 having physiologically acceptable excipients. As used herein, “physiologically acceptable excipients” include, but are not limited to, any adjuvants, carriers, diluents, preservatives, dispersants, suspending agents, stabilizers, isotonic agents, solvents, surfactants, or emulsifiers that are acceptable for use in humans or livestock. In some embodiments, the CAR-T cells of the present invention or a pharmaceutical composition containing them are used to treat DLL3-related disorders in a subject. Accordingly, a method for treating a DLL3-related disorder is provided, 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-related disorder. The “therapeutally effective amount” of the antibody, CAR-T cells, or pharmaceutical composition may vary depending on factors such as the subject's (e.g., patient's) condition, age, sex, and weight. The term “therapeutic dose” may include the amount that is effective in “treating” the subject. When a therapeutic dose is specified, the exact amount administered as intended in a particular embodiment may be determined by a physician in consideration of the subject’s condition. In some embodiments, DLL3-related disorders are cancers that express 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-related disorder is lung cancer, particularly small cell lung cancer (SCLC). [Examples]

[0128] The examples described herein are not intended to represent all or only experiments that have been conducted. While efforts have been made to ensure accuracy of the numerical values ​​used (e.g., quantities, temperatures, etc.), some experimental error and deviation should be taken into consideration. Unless otherwise specified, parts are parts by weight, molecular weight is weight-average molecular weight, temperature is degrees Celsius, and pressure is atmospheric pressure or near it.

[0129] Example 1. Animal immunization and antibody library construction In this example, we demonstrated that immune camels showed a good immune response to human or rhesus monkey DLL3 protein, and that acquired immune libraries exhibited excellent quality.

[0130] animal immunity An immunogen containing the extracellular domain of human DLL3 protein (aa27~466) (AdipoGen,AG-40B-0151) with an N-terminal FLAG tag or / and DLL3 expression plasmid or DLL3-expressing cells (CHO-K1 / DLL3 or / and CHO-K1 / EGF4) was mixed with an adjuvant or PBS and injected into camels. Typically, camels were immunized 2 to 4 times at 1 to 2-week intervals. After multiple immunizations, the immune response to the target antigen DLL3 was evaluated by serum titration through both enzyme-linked immunosorbent assay (ELISA) and flow cytometry assays.

[0131] Phage display library construction Total RNA was extracted from immunocamelli lymphocytes using TRIZOL® reagents, following the manufacturer's protocol. cDNA was synthesized based on the RNA template using the PRIMESCRIPT® First Strand cDNA Synthesis Kit and oligo(dT)20 primers, following the manufacturer's protocol. H To create an H phage library, V phage is obtained from camel cDNA. H H was amplified.

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

[0133] Phage display A phage display library was constructed using sdAb obtained by immunoassay (human DLL3 protein (aa27~466) with an N-terminal FLAG tag (AdipoGen, AG-40B-0151) or / and an immunogen containing the extracellular domain of a DLL3 expression plasmid or DLL3-expressing cells (CHO-K1 / DLL3 or / and CHO-K1 / EGF4)). Another human Fab phage display library was synthesized. For further use, both phage libraries were rescued and stored after filter sterilization at 4°C. Binding phages were isolated from the two phage libraries using protein-based and cell-based panning. Using both libraries, at least one panning run was performed using both protein-based and cell-based panning methods until the percentage of DLL3-specific phage clones reached 30%. The output phages were evaluated for each run in terms of the total number of output clones, the percentage of DLL3-positive clones by ELISA, and the sequence diversity of DLL3-specific clones. Based on these results, we selected the best panning output for high-throughput screening.

[0134] High-throughput screening Selected output phages were used to infect exponentially growing E. coli (E. coli) cells. Double-stranded DNA from the output phages was extracted. For high-throughput screening, sdAb / Fab inserts were cleaved from phagemid vectors and inserted into antibody fragment expression vectors. The resulting plasmids were used to transform exponentially growing E. coli (E. coli) cells, which were then seeded and grown overnight at 37°C. Thousands of colonies were individually selected and grown in 96-deep-well plates containing 1 mL of 2×YT medium. Antibody fragment expression was induced by adding 1 mM IPTG.

[0135] The sdAb / Fab proteins in the supernatant were analyzed by ELISA for their ability to bind to the DLL3 ECD protein, and by FACS for their ability to bind to the DLL3-expressing SHP-77 cell line (American Type Culture Collection (ATCC) (registered trademark) CRL-2195 (trademark)) and CHO-K1 / human DLL3 (home-developed). All binders were sequenced. Redundant sequences were removed. A total of 81 camel sdAbs and 2 human Fab binders were obtained that bound to both human and rhesus monkey DLL3 proteins and cell lines. All of 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 conducted as follows: Crude sdAb / Fab protein was captured on a sensor chip using an affinity tag. A high concentration (100 nM) of human DLL3 flowed over the sensor chip surface and bound to the antibody fragment for 300 seconds, followed by injection of running buffer to dissociate the formed complex. The on-rate (ka) and off-rate (kd) were estimated based on a single association and dissociation curve, and these were used to determine the equilibrium dissociation constant (K). D The binding affinity of some of these unique binders was estimated. The binding affinity of some of these unique binders is shown in Table 7.

[0137] Table 1 lists the CDR sequences of anti-DLL3 camel sdAb, and Table 2 lists the CDR sequences of anti-DLL3 human scFv.

[0138] (Table 1) CDR sequences of anti-DLL3 camel sdAb TIFF0007894963000001.tif118151TIFF0007894963000002.tif242151TIFF0007894963000003.tif243151TIFF0007894963000004.tif208151

[0139] (Table 2) CDR sequences of anti-DLL3 human scFv TIFF0007894963000005.tif51151

[0140] The amino acid sequences of anti-DLL3 sdAb are listed in Table 3. The CDR of sdAb is underlined. Nucleic acid sequences encoding anti-DLL3 sdAb are shown in SEQ ID NOs. 368-448.

[0141] (Table 3) Amino acid sequence of anti-DLL3 camel sdAb TIFF0007894963000006.tif190150TIFF0007894963000007.tif245150TIFF0007894963000008.tif245150TIFF00078949630 00009.tif241150TIFF0007894963000010.tif245150TIFF0007894963000011.tif246150TIFF0007894963000012.tif102150

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

[0143] (Table 4) Amino acid sequence of anti-DLL3 human scFv TIFF0007894963000013.tif95147

[0144] Example 3. Preparation of a single-specific camel CAR The amino acid sequences of the anti-DLL3 camel sdAb fragments are shown in Table 3 above, and the nucleic acid sequences of the anti-DLL3 camel sdAb fragments are shown as SEQ ID NOs. 368-448. CAR constructs (SEQ ID NOs. 476-484) were constructed using the sdAb fragments and additional sequences from Table 3. A reference CAR construct (CAR3) was also constructed using CAR3 scFv (SEQ ID NOs. 473), a human anti-DLL3 scFv. The full-length CAR contains, from the N-terminus to the C-terminus, a CD8α signal peptide (SEQ ID NOs. 465), the DLL3-binding domain sdAb shown 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. Next, the nucleic acid encoding the CAR fragment was cloned into a lentiviral vector, and a full-length CAR construct was created in a single coding frame using a human EF1α promoter for expression. The resulting CAR skeleton vector was named "PLLV-hEF1α-DLL3".

[0145] Example 4. Production of camel anti-DLL3 CAR-T cells Preparation of lentiviruses A lentiviral packaging plasmid mixture containing pCMV-ΔR-8.47 and pMD2.G (Addgene, catalog number 12259) was pre-mixed with PLLV-hEF1α-DLL3 vector in the presence of polyethyleneimine in a pre-optimized ratio (1:1:1:2) before being added 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 virus pellet was rinsed with pre-cooled DPBS. The virus was appropriately aliquoted and immediately stored at -80°C. Viral titer was determined by measuring the transduction efficiency to supT1 cell lines 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 (trademark) PLUS medium according to the manufacturer's protocol. Human T cells were purified from PMBCs using a pan-T cell isolation kit (Miltenyi, catalog number 130-096-535) according to the manufacturer's protocol. Subsequently, the purified T cells were pre-activated 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. Lentiviral stock was transduced into the pre-activated T cells in the presence of 7 μg / mL of polybrene. Next, the transduced cells were transferred to a cell culture incubator and the transgene was expressed under suitable conditions.

[0147] Example 5. Evaluation of the in vitro activity of camel anti-DLL3 CAR-T cells In vitro cytotoxicity assay On the 6th day after transduction, the transduced T cells were collected and co-incubated with the DLL3-expressing tumor cell line SHP-77 for 20 hours at effector (CAR-T) to target cell ratios of 2:1 and 5:1. In all assays, CAR3 CAR-T cells were used as a reference to compare assay variability and / or to 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 a lactate dehydrogenase (LDH) assay. The results show that CAR3 CAR-T and some anti-DLL3 CAR-T exhibit strong anti-tumor activity against SHP-77 cells in vitro, while UnT has no target cell killing effect (Figure 2).

[0149] IFN-γ and TNF-α release detection In addition, supernatants from in vitro cytotoxicity assays were collected and CAR-induced cytokine release, such as interferon gamma (IFN-γ) and TNF-α release, was evaluated. As shown in FIGS. 3A and 3B, CAR3 CAR-T and some anti-DLL3 CAR-T were stimulated by SHP-77 and produced IFN-γ and TNF-α, while UnT produced little IFN-γ and TNF-α. For the protocol of IFN-γ and TNF-α release detection, refer to the human TNF-α kit and IFN-γ kit of CISBIO.

[0150] CAR-T Expansion and Proliferation by Long-Term Stimulation Assay On day 0, 1×10 5 individual SHP-77 cells were seeded into 24-well plates to establish a monolayer. On day 1, the transduced T cells were counted, and 2×10 5 0 + viable CAR 5 T cells were seeded on top of the SHP-77 cells in fresh medium without cytokines. On day 3, a new 1×10 5 individual SHP-77 cell monolayer was seeded on top of the CAR-T cells. On day 4, for each well, the viable CAR-T cells were counted. On the same day, 2×10 + individual, expanded wells (having at least this amount of cells) of CAR + T cells were reseeded to establish a new monolayer as on day 1. The process was repeated and stimulated 3 - 4 times. The expansion ratio after each stimulation was calculated as [viable CAR 5 T cells on day 4] / 2×10

[0151] individual (the amount of CAR T cells seeded on day 1 of each stimulation). To normalize for the cells discarded in each new stimulation, the cumulative expansion ratio was determined by [(expansion ratio)×(expansion ratio + 1)...].

[0152] Example 6. Evaluation of the in vivo efficacy of camel CAR by inhibiting CAR-T cell-mediated tumor growth. The antitumor activity of camel CAR was evaluated in an SHP-77 tumor model. SHP-77 cells were subcutaneously transplanted into NOD / SCID mice and randomized into 7 groups (4 mice per group, day 0). Group 1: Vegetative state (PBS only); Group 2: UnT (negative control); Group 3: CAR3; Group 4: CAS64380; Group 5: CAS64511; Group 6: CAS63931; Group 7: CAS63997. When the tumor was palpable (100 mm) 3 ), initiate treatment with CAR-T cells, UnT cells, or a medium (PBS only), when the tumor volume is approximately 3000 mm³. 3 When the tumor volume reached a certain level, the mice were euthanized. Tumor volume was measured twice a week. CAR-T cells were injected at a rate of 1 × 10⁶. 6 Individual CAR-positive T cells / mouse were administered intravenously. After tumor cell transplantation, the mice and tumors were monitored for approximately 21 days.

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

[0154] Example 7. Humanization of camel sdAb Selected camel sdAbs (SEQ ID NOs. 279, 294, 297, 312) were humanized using CDR transplantation technology (see, for example, U.S. Patent No. 5,225,539). Specifically, the camel sdAb sequences were compared to those available in the Research Collaboratory for Structural Bioinformatics (RCSB) Protein Databank. Homology models for each camel sdAb were constructed based on the closest VH structure. From the model structures, residues located near the CDR or embedded internally within the molecule (i.e., those with less than 15% of the surface area accessible to the side-chain solvent) were identified.

[0155] Subsequently, each camel sdAb sequence was subjected to a BLAST search 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, for example, 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 transplantation method, the CDR of the human acceptor was replaced with the CDR of the camel sdAb to produce a straight-graft sequence. Straight-graft antibodies typically lose their binding activity, which needs to be restored by substituting framework residues, which are critical to antibody activity, with non-human residues. Amino acid residues located near the CDR or embedded within the molecule are typically important for antibody activity and structure, and therefore needed to be potential reverse mutation sites. A series of humanized mutants were designed using this method. The CDR amino acid sequences of the humanized mutants are shown in Table 5. The full-length amino acid sequences of the humanized mutants are shown in Table 6. The CDR is underlined.

[0156] (Table 5) CDR sequences of humanized anti-DLL3 sdAb TIFF0007894963000014.tif124151

[0157] (Table 6) Amino acid sequence of anti-DLL3 humanized sdAb TIFF0007894963000015.tif234147

[0158] Camel and humanized sdAb sequences were fused with human IgG1 hinge and Fc to obtain chimeric and humanized HCAb sequences. DNA encoding these HCAb was synthesized and inserted into a pTT5 vector. HEK293 cells were transfected using an HCAb expression plasmid. Crude HCAb proteins secreted into culture medium were subjected to SPR affinity measurement as follows: The anti-human Fc pAb (GE Healthcare) antibody to be captured was immobilized on a Biacore® CM5 chip using EDC-activated amine coupling chemistry to approximately 6,000 RU. The target HCAb was captured on the sensor chip surface for 300 seconds. Human DLL3 (AdipoGen, AG-40B-0151) was flowed onto the sensor chip surface in a series of increasing concentrations. The association and dissociation phases were monitored. Captured antibodies and antigens were removed between cycles using a 10 mM glycine-HCl buffer, pH 2.0, and a new binding surface for the antigen was identified. Using a 1:1 coupling model, the resulting sensorgram is fitted overall to determine the 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 camel sdAbs (Table 7). The majority of humanized antibodies retained their binding affinity to the camel sdAbs. This example demonstrates the success of humanization of sdAbs using our standard protocol. The majority of sdAbs retained their binding affinity after humanization.

[0160] scFv, when assayed using the same procedure as above, yields equivalent K D It has a value (AS56788 and AS56704 in Table 7).

[0161] (Table 7) Monovalent binding affinity of camel, humanized antibodies, and scFv TIFF0007894963000016.tif126133

[0162] Example 8. Preparation of a single-specific humanized CAR The amino acid sequence of anti-DLL3 humanized sdAb is shown in Table 6 above, and the nucleic acid sequence of anti-DLL3 humanized sdAb is listed as SEQ ID NOs: 449-461. Complete CAR constructs (SEQ ID NOs: 485-494) were constructed using the humanized sdAb and additional sequences in Table 6. The full-length CAR has, from the N-terminus to the C-terminus, a CD8α signal peptide (SEQ ID NO: 465), the DLL3 binding domain (humanized sdAb) shown 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. Next, the nucleic acids encoding the CAR fragments were cloned into lentiviral vectors, and full-length CAR constructs were created in a single coding frame using a human EF1α promoter for expression. The obtained CAR skeleton vector was named "PLLV-hEF1α-DLL3".

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

[0164] In vitro cytotoxicity assay The results are shown in Figures 6A to 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 and CAR-induced cytokine release, such as interferon-gamma (IFN-γ) and tumor necrosis factor-α (TNF-α) release, was evaluated. As shown in Figures 7A and 7B, CAR3 CAR-T and some anti-DLL3 CAR-T cells produced IFN-γ and TNF-α when stimulated by SHP-77, while UnT cells produced little to no IFN-γ or TNF-α.

[0166] CART Expansion and Proliferation by Long-Term Stimulation Assay On day 0, 1×10 5 SHP-77 cells were seeded in 24-well plates to establish a monolayer. On day 1, CAR-T cells were counted, and 2×10 5 viable CAR + T cells were seeded on top of the SHP-77 cells in fresh medium without cytokines. On day 3, a new 1×10 5 NCI-H82 cell monolayer was seeded 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 CAR + T cells from the expanded wells (having at least this amount of cells) were reseeded to establish a new monolayer as on day 1. The process was repeated and stimulated 3 - 4 times. The expansion ratio after each stimulation was calculated as [viable CAR + T cells on day 4] / 2×10 5 cells (the amount of CAR-T cells seeded on day 1 of each stimulation). To normalize for the cells discarded in each new stimulation, the cumulative expansion ratio was determined by [(expansion ratio)×(expansion ratio + 1)...].

[0167] After 3 stimulations, the expansion ratios of different CAR-T constructs were calculated. As shown in Figure 8, most CAR-T constructs expanded more than CAR3 CAR-T after 3 stimulations by SHP-77 tumor cells.

[0168] Example 10. In Vivo Evaluation of the Antitumor Activity of a Humanized CAR by Inhibiting Tumor Growth Mediated by CAR-T Cells The antitumor activity of the humanized CAR was evaluated in a SHP-77 tumor model. SHP-77 cells were subcutaneously transplanted 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, on day ). When the tumor was palpable (100 mm 3), initiate treatment with CAR-T cells, UnT cells, or a medium (PBS only), when the tumor volume is approximately 3000 mm³. 3 When it reached this point, the mice were euthanized. Tumor volume was measured twice a week. CAR-T cells were 0.2 × 10⁶ 6 Individual CAR-positive T cells / mouse were administered intravenously. After tumor cell transplantation, the mice and tumors were monitored for approximately 21 days.

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

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

[0171] Some amino acid sequences and nucleic acid sequences described in this specification are listed below. Camel sdAb nucleic acid sequence Sequence ID 368 (Camel sdAb AS63930 nucleic acid sequence) TIFF0007894963000017.tif36147 Sequence ID 369 (Camel sdAb AS63932 nucleic acid sequence) TIFF0007894963000018.tif36146 Sequence ID 370 (Camel sdAb AS63951 nucleic acid sequence) TIFF0007894963000019.tif36146 Sequence ID 371 (Camel sdAb AS63984 nucleic acid sequence) TIFF0007894963000020.tif36146 Sequence ID 372 (Camel sdAb AS63987 nucleic acid sequence) TIFF0007894963000021.tif36146 Sequence ID 373 (Camel sdAb AS63997 nucleic acid sequence) TIFF0007894963000022.tif36146 Sequence ID 374 (Camel sdAb AS64047 nucleic acid sequence) TIFF0007894963000023.tif36146 Sequence ID 375 (Camel sdAb AS64052 nucleic acid sequence) TIFF0007894963000024.tif36146 Sequence ID 376 (Camel sdAb AS64062 nucleic acid sequence) TIFF0007894963000025.tif36146 Sequence ID 377 (Camel sdAb AS64072 nucleic acid sequence) TIFF0007894963000026.tif36147 Sequence ID 378 (Camel sdAb AS64097 nucleic acid sequence) TIFF0007894963000027.tif36146 Sequence ID 379 (Camel sdAb AS64114 nucleic acid sequence) TIFF0007894963000028.tif36146 Sequence ID 380 (Camel sdAb AS64123 nucleic acid sequence) TIFF0007894963000029.tif36146 Sequence ID 381 (Camel sdAb AS64130 nucleic acid sequence) TIFF0007894963000030.tif36147 Sequence ID 382 (Camel sdAb AS64137 nucleic acid sequence) TIFF0007894963000031.tif36147 Sequence ID 383 (Camel sdAb AS64142 nucleic acid sequence) TIFF0007894963000032.tif36146 Sequence ID 384 (Camel sdAb AS64154 nucleic acid sequence) TIFF0007894963000033.tif36146 Sequence ID 385 (Camel sdAb AS64160 nucleic acid sequence) TIFF0007894963000034.tif36146 Sequence ID 386 (Camel sdAb AS64228 nucleic acid sequence) TIFF0007894963000035.tif37146 Sequence ID 387 (Camel sdAb AS64300 nucleic acid sequence) TIFF0007894963000036.tif36147 Sequence ID 388 (Camel sdAb AS64380 nucleic acid sequence) TIFF0007894963000037.tif36147 Sequence ID 389 (Camel sdAb AS64395 nucleic acid sequence) TIFF0007894963000038.tif36146 Sequence ID 390 (Camel sdAb AS64443 nucleic acid sequence) TIFF0007894963000039.tif36146 Sequence ID 391 (Camel sdAb AS64511 nucleic acid sequence) TIFF0007894963000040.tif36146 Sequence ID 392 (Camel sdAb AS64536 nucleic acid sequence) TIFF0007894963000041.tif36146 Sequence ID 393 (Camel sdAb AS64597 nucleic acid sequence) TIFF0007894963000042.tif36146 Sequence ID 394 (Camel sdAb AS64617 nucleic acid sequence) TIFF0007894963000043.tif36146 Sequence ID 395 (Camel sdAb AS64634 nucleic acid sequence) TIFF0007894963000044.tif36146 Sequence ID 396 (Camel sdAb AS69498 nucleic acid sequence) TIFF0007894963000045.tif36146 Sequence ID 397 (Camel sdAb AS69500 ​​nucleic acid sequence) TIFF0007894963000046.tif36147 Sequence ID 398 (Camel sdAb AS69527 nucleic acid sequence) TIFF0007894963000047.tif36146 Sequence ID 399 (Camel sdAb AS68280 nucleic acid sequence) TIFF0007894963000048.tif36146 Sequence ID 400 (Camel sdAb AS68355 nucleic acid sequence) TIFF0007894963000049.tif36147 Sequence ID 401 (Camel sdAb AS69443 nucleic acid sequence) TIFF0007894963000050.tif36147 Sequence ID 402 (Camel sdAb AS75376 nucleic acid sequence) TIFF0007894963000051.tif36146 Sequence ID 403 (Camel sdAb AS75387 nucleic acid sequence) TIFF0007894963000052.tif37147 Sequence ID 404 (Camel sdAb AS75695 nucleic acid sequence) TIFF0007894963000053.tif36146 Sequence ID 405 (Camel sdAb AS76169 nucleic acid sequence) TIFF0007894963000054.tif36146 Sequence ID 406 (Camel sdAb AS63931 nucleic acid sequence) TIFF0007894963000055.tif36147 Sequence ID 407 (Camel sdAb AS63937 nucleic acid sequence) TIFF0007894963000056.tif36146 Sequence ID 408 (Camel sdAb AS63948 nucleic acid sequence) TIFF0007894963000057.tif36146 Sequence ID 409 (Camel sdAb AS63956 nucleic acid sequence) TIFF0007894963000058.tif36147 Sequence ID 410 (Camel sdAb AS63965 nucleic acid sequence) TIFF0007894963000059.tif36146 Sequence ID 411 (Camel sdAb AS63993 nucleic acid sequence) TIFF0007894963000060.tif36146 Sequence ID 412 (Camel sdAb AS63999 nucleic acid sequence) TIFF0007894963000061.tif36146 Sequence ID 413 (Camel sdAb AS64006 nucleic acid sequence) TIFF0007894963000062.tif36146 Sequence ID 414 (Camel sdAb AS64057 nucleic acid sequence) TIFF0007894963000063.tif36147 Sequence ID 415 (Camel sdAb AS64060 nucleic acid sequence) TIFF0007894963000064.tif36146 Sequence ID 416 (Camel sdAb AS64071 nucleic acid sequence) TIFF0007894963000065.tif36146 Sequence ID 417 (Camel sdAb AS64093 nucleic acid sequence) TIFF0007894963000066.tif36146 Sequence ID 418 (Camel sdAb AS64118 nucleic acid sequence) TIFF0007894963000067.tif36146 Sequence ID 419 (Camel sdAb AS64120 nucleic acid sequence) TIFF0007894963000068.tif42146 Sequence ID 420 (Camel sdAb AS64124 nucleic acid sequence) TIFF0007894963000069.tif36146 Sequence ID 421 (Camel sdAb AS64135 nucleic acid sequence) TIFF0007894963000070.tif36147 Sequence ID 422 (Camel sdAb AS64163 nucleic acid sequence) TIFF0007894963000071.tif36146 Sequence ID 423 (Camel sdAb AS64182 nucleic acid sequence) TIFF0007894963000072.tif36146 Sequence ID 424 (Camel sdAb AS64183 nucleic acid sequence) TIFF0007894963000073.tif36146 Sequence ID 425 (Camel sdAb AS64207 nucleic acid sequence) TIFF0007894963000074.tif36146 Sequence ID 426 (Camel sdAb AS64276 nucleic acid sequence) TIFF0007894963000075.tif36146 Sequence ID 427 (Camel sdAb AS64336 nucleic acid sequence) TIFF0007894963000076.tif36146 Sequence ID 428 (Camel sdAb AS64346 nucleic acid sequence) TIFF0007894963000077.tif36146 Sequence ID 429 (Camel sdAb AS64420 nucleic acid sequence) TIFF0007894963000078.tif36146 Sequence ID 430 (Camel sdAb AS64473 nucleic acid sequence) TIFF0007894963000079.tif36146 Sequence ID 431 (Camel sdAb AS64475 nucleic acid sequence) TIFF0007894963000080.tif36146 Sequence ID 432 (Camel sdAb AS64513 nucleic acid sequence) TIFF0007894963000081.tif36146 Sequence ID 433 (Camel sdAb AS64562 nucleic acid sequence) TIFF0007894963000082.tif36146 Sequence ID 434 (Camel sdAb AS64583 nucleic acid sequence) TIFF0007894963000083.tif36146 Sequence ID 435 (Camel sdAb AS64594 nucleic acid sequence) TIFF0007894963000084.tif36146 Sequence ID 436 (Camel sdAb AS64605 nucleic acid sequence) TIFF0007894963000085.tif36146 Sequence ID 437 (Camel sdAb AS64606 nucleic acid sequence) TIFF0007894963000086.tif36146 Sequence ID 438 (Camel sdAb AS68121 nucleic acid sequence) TIFF0007894963000087.tif36147 Sequence ID 439 (Camel sdAb AS68170 nucleic acid sequence) TIFF0007894963000088.tif36147 Sequence ID 440 (Camel sdAb AS63964 nucleic acid sequence) TIFF0007894963000089.tif36146 Sequence ID 441 (Camel sdAb AS64116 nucleic acid sequence) TIFF0007894963000090.tif36146 Sequence ID 442 (Camel sdAb AS68270 nucleic acid sequence) TIFF0007894963000091.tif36147 Sequence ID 443 (Camel sdAb AS68320 nucleic acid sequence) TIFF0007894963000092.tif36147 Sequence ID 444 (Camel sdAb AS68351 nucleic acid sequence) TIFF0007894963000093.tif36147 Sequence ID 445 (Camel sdAb AS75378 nucleic acid sequence) TIFF0007894963000094.tif36146 Sequence ID 446 (Camel sdAb AS75383 nucleic acid sequence) TIFF0007894963000095.tif36147 Sequence ID 447 (Camel sdAb AS75751 nucleic acid sequence) TIFF0007894963000096.tif36146 Sequence ID 448 (Camel sdAb AS76422 nucleic acid sequence) TIFF0007894963000097.tif36146

[0172] Humanized camel sdAb nucleic acid sequence Sequence ID 449 (Humanized sdAb AS64380VH4 nucleic acid sequence) TIFF0007894963000098.tif36146 Sequence ID 450 (Humanized sdAb AS64380VH5 nucleic acid sequence) TIFF0007894963000099.tif36146 Sequence ID 451 (Humanized sdAb AS64380VH6 nucleic acid sequence) TIFF0007894963000100.tif36146 Sequence ID 452 (Humanized sdAb AS64380VH7 nucleic acid sequence) TIFF0007894963000101.tif36146 Sequence ID 453 (Humanized sdAb AS64511VH4 nucleic acid sequence) TIFF0007894963000102.tif36147 Sequence ID 454 (Humanized sdAb AS64511VH5 nucleic acid sequence) TIFF0007894963000103.tif36147 Sequence ID 455 (Humanized sdAb AS64511VH6 nucleic acid sequence) TIFF0007894963000104.tif36147 Sequence ID 456 (Humanized sdAb AS63931VH4 nucleic acid sequence) TIFF0007894963000105.tif36146 Sequence ID 457 (Humanized sdAb AS63931VH5 nucleic acid sequence) TIFF0007894963000106.tif36146 Sequence ID 458 (Humanized sdAb AS63931VH6 nucleic acid sequence) TIFF0007894963000107.tif36146 Sequence ID 459 (Humanized sdAb AS63997VH4 nucleic acid sequence) TIFF0007894963000108.tif36147 Sequence ID 460 (Humanized sdAb AS63997VH5 nucleic acid sequence) TIFF0007894963000109.tif36147 Sequence ID 461 (Humanized sdAb AS63997VH6 nucleic acid sequence) TIFF0007894963000110.tif36147 Sequence ID 462 (Linker amino acid sequence) TIFF0007894963000111.tif3128 Sequence ID 463 (Linker amino acid sequence) TIFF0007894963000112.tif3128 Sequence ID 464 (Linker amino acid sequence) TIFF0007894963000113.tif3128 Sequence ID No. 465 (CD8α signal peptide amino acid sequence) TIFF0007894963000114.tif3128 Sequence ID 466 (CD8α hinge amino acid sequence) TIFF0007894963000115.tif9146SEQ ID NO: 467 (CD8α transmembrane domain amino acid sequence) TIFF0007894963000116.tif3128 Sequence ID 468 (4-1BB intracellular domain amino acid sequence) TIFF0007894963000117.tif4128 Sequence ID No. 469 (CD28 intracellular domain amino acid sequence) TIFF0007894963000118.tif4128 Sequence ID 470 (CD3ζ intracellular domain amino acid sequence) TIFF0007894963000119.tif9147 Sequence ID 471 (F2A element amino acid sequence) TIFF0007894963000120.tif4128 Sequence ID 472 (P2A element amino acid sequence) TIFF0007894963000121.tif4128 Sequence ID 473 (CAR3 anti-DLL3 scFv amino acid sequence) TIFF0007894963000122.tif26147 Sequence ID No. 474 (CD28 transmembrane domain amino acid sequence) TIFF0007894963000123.tif3128 Sequence ID 475 (CD28 Hinge) TIFF0007894963000124.tif3128

[0173] Camel anti-DLL3 CAR sequence Sequence ID 476 (CAS63997) TIFF0007894963000125.tif37145 Sequence ID 477 (CAS64380) TIFF0007894963000126.tif37146 Sequence ID 478 (CAS64511) TIFF0007894963000127.tif37147 Sequence ID 479 (CAS64617) TIFF0007894963000128.tif35147 Sequence ID 480 (CAS69443) TIFF0007894963000129.tif37147 Sequence ID 481 (CAS63931) TIFF0007894963000130.tif37145 Sequence ID 482 (CAS64047) TIFF0007894963000131.tif37147 Sequence ID 483 (CAS64052) TIFF0007894963000132.tif37146 Sequence ID 484 (CAS64062) TIFF0007894963000133.tif37147

[0174] Humanized anti-DLL3 CAR sequence Sequence ID 485 (CAS64380VH4) TIFF0007894963000134.tif37145 Sequence ID 486 (CAS64380VH5) TIFF0007894963000135.tif37145 Sequence ID 487 (CAS64380VH6) TIFF0007894963000136.tif37146 Sequence ID 488 (CAS64380VH7) TIFF0007894963000137.tif37146 Sequence ID 489 (CAS64511VH4) TIFF0007894963000138.tif37146 Sequence ID 490 (CAS64511VH5) TIFF0007894963000139.tif37147 Sequence ID 491 (CAS64511VH6) TIFF0007894963000140.tif37146 Sequence ID 492 (CAS63997VH4) TIFF0007894963000141.tif37146 Sequence ID 493 (CAS63997VH5) TIFF0007894963000142.tif37146 Sequence ID 494 (CAS63997VH6) TIFF0007894963000143.tif37146

[0175] Anti-DLL3 human scFv VL and VH domain nucleic acid sequences Sequence ID 511 (Nucleic acid sequence in the VL domain of anti-DLL3 human scFv AS56704) TIFF0007894963000144.tif31146 Sequence ID 512 (Nucleic acid sequence in the VH domain of anti-DLL3 human scFv AS56704) TIFF0007894963000145.tif36146 Sequence ID 513 (Nucleic acid sequence in the VL domain of anti-DLL3 human scFv AS56788) TIFF0007894963000146.tif31146 Sequence ID 514 (Nucleic acid sequence in the VH domain of anti-DLL3 human scFv AS56788) TIFF0007894963000147.tif36146

[0176] Human anti-DLL3 scFv CAR sequence Sequence ID 515 (CAS 56704) TIFF0007894963000148.tif48147 Sequence ID 516 (CAS 56788) TIFF0007894963000149.tif48147

[0177] Anti-DLL3 benchmark CAR Sequence ID 517 (1H2.1 amino acid sequence) TIFF0007894963000150.tif42146

[0178] Example 11. Evaluation of in vitro activity of humanized anti-DLL3 tandem CAR-T cells To improve the antitumor efficacy of CAR-T receptors, the inventors constructed three tandem CARs (T1, T2, and T3). The amino acid sequences of the tandem CARs are presented in SEQ ID NOs. 518-520. The amino acid sequences of anti-DLL3 humanized sdAb fragments are presented in SEQ ID NOs. 356 (AS64380VH5) and 366 (AS63997VH5). A reference CAR construct was also prepared using the anti-DLL3 CAR 1H2.1 (SEQ ID NO. 517, see, for example, International Publication No. 2019200007). The full-length CAR has, from the N-terminus to the C-terminus, a DLL3-binding domain sdAb presented by the CD8α signal peptide (SEQ ID NO: 465), 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. In the case of T1, both sdAb1 and sdAb2 were AS64380VH5. In the case of T2, both sdAb1 and sdAb2 were AS63997VH5. In the case of T3, sdAb1 and sdAb2 were AS63997VH5 and AS64380VH5, respectively. Next, the nucleic acid encoding the CAR fragment was cloned into a lentiviral vector, and a full-length CAR construct was created in a single coding frame using a human EF1α promoter for expression.

[0179] Sequence ID 518 (T1 amino acid sequence) TIFF0007894963000151.tif48147 Sequence ID 519 (T2 amino acid sequence) TIFF0007894963000152.tif48146 Sequence ID 520 (T3 amino acid sequence) TIFF0007894963000153.tif48146

[0180] In vitro cytotoxicity assay Nine days after transduction, transduced T cells were collected and co-incubated for 22 hours 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 cell lines (NCI-H460 and HEK293) at effector (CAR-T) to target cell ratios of 0.5:1 and 2:1. In all assays, CAR3 CAR-T cells were used as a reference to compare assay variability and / or as a control. Untransduced T cells (UnT) were used as a negative control.

[0181] The cytotoxicity of transduced T cells was determined by lactate dehydrogenase (LDH) assay. The results showed that CAR3 CAR-T and some anti-DLL3 tandem CAR-T cells exhibited potent antitumor activity against SHP-77 cells in vitro, while UnT cells did not exhibit the target cell-killing effect (Figure 11A-C), and DLL3-negative expressing cells (NCI-H460 and HEK293) did not induce cytotoxicity (Figure 11D-E). In addition to CAR3, the inventors also compared the in vitro cytotoxicity of T3 and 1H2.1 against SHP-77 cells. The results showed that T3 cells exhibited comparable or less potent cell-killing activity in short-term stimulation (Figure 11V).

[0182] IFN-γ and TNF-α emission detection In addition, supernatants from in vitro cytotoxicity assays were collected and CAR-induced cytokine release, such as interferon-gamma (IFN-γ) and TNF-α release, was evaluated. As shown in Figures 11F-K, CAR3 CAR-T and some anti-DLL3 tandem CAR-T cells produced IFN-γ and TNF-α when stimulated by DLL3-expressing cell lines, while UnT cells produced almost no IFN-γ and TNF-α. DLL3-negative cell lines did not reduce the specific release of IFN-γ and TNF-α (data not shown). For protocols to detect IFN-γ and TNF-α release, refer to the CISBIO human TNF-α kit and IFN-γ kit.

[0183] Compared to 1H2.1, T3 released more IFN-γ and TNF-α (after 22 hours of co-incubation) (Figure 11W~X).

[0184] Tandem CAR-T cell toxicity and proliferation by long-term stimulation assay DLL3 CAR-T cells were evaluated using repeated antigen stimulation assays. Upon repeated stimulation with SCLC cell lines and control cell lines, tandem CAR-T cells T3 showed more potent cytotoxicity against SCLC cells, particularly SHP-77 and NCI-H82 cells (Figure 11L-P). In addition to cytotoxic activity, tandem CAR-T cells T3 also showed higher proliferative capacity than other CARTs, especially when stimulated with SHP-77 and NCI-H82 cells (Figure 11Q-U).

[0185] In addition to CAR3, the inventors also compared the in vitro cytotoxicity of T3 and 1H2.1 against SHP-77 cells. The results showed that T3 exhibited superior cytotoxicity and proliferative effects under prolonged stimulation (Figure 11Y-Z).

[0186] The repeated stimulation was performed as follows: Round 1: CAR-T cells and 3 × 10 5 Individual target cells (e.g., SHP-77) were added to 24-well plates in a 1:5 effector-to-target cell ratio and co-incubated for 3 days in a carbon dioxide incubator at 37°C and 5% CO2. For cytokine detection, 200 μL of cell culture supernatant was pipetteed, the co-incubated cells were collected, and %CD3 and CAR positivity rates were evaluated by flow cytometry. Round 2: Based on the CAR-T positivity rate of cells collected in Round 1, the collected cells were further co-incubated for 3 days with the same volume of novel target cells (SHP-77) in a 1:2 effector-to-target cell ratio. For cytokine detection, 200 μL of cell culture supernatant was pipettered, the co-incubated cells were collected, and %CD3 and CAR positivity rates were evaluated by flow cytometry. Round 3 and subsequent rounds were conducted in the same manner as Round 2, based on the CAR-T positive rate of each preceding round.

[0187] Example 12. Evaluation of in vitro activity of humanized anti-DLL3 CAR-T cells armed with PD-1 DNR or CSR. To improve the persistence of CAR-T receptors, the inventors constructed DLL3 CARs armed with either 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 shown in SEQ ID NOs. 521-522. The PD-1DNR and PD-1CSR sequences were ligated to the C-terminus of T3 via P2A. The amino acid sequences of PD-1DNR and PD-1CSR are shown in SEQ ID NOs. 523-524. A schematic diagram of the CAR construct is shown in Figure 10B.

[0188] Sequence ID 521 (T3-PD-1DNR amino acid sequence) TIFF0007894963000154.tif75146 Sequence ID 522 (T3-PD-1CSR amino acid sequence) TIFF0007894963000155.tif70146 Sequence ID 523 (PD-1DNR amino acid sequence) TIFF0007894963000156.tif26146 Sequence ID 524 (PD-1CSR amino acid sequence) TIFF0007894963000157.tif20146

[0189] DLL3 CAR-T cells armed with PD-1DNR or PD-1CSR were evaluated using repeated antigen stimulation assays. When repeatedly stimulated with SHP-77 cells, the armed CAR-T cells did not show enhanced cytotoxicity or improved proliferative capacity compared to conventional CAR-T cells (Figure 12A-B). When repeatedly stimulated with SHP-77 / PD-L1 cells (SHP-77 cells overexpressing human PD-L1), CAR-T cells armed with PD-1 CSR showed superior cytotoxicity and proliferative capacity (Figure 12C-D).

[0190] Example 13. TGF-β-DNR enhances the antitumor efficacy of DLL3 CAR-T cells. Construction of DLL3 CAR-T cells armed with TGF-β-DNR To improve the antitumor performance of DLL3 CAR-T cells within the tumor microenvironment, the TGF-β-DNR sequence was incorporated into DLL3 CARs, as shown in Figure 13A. TGF-β-DNR is a cleaved version of TGFBRII, consisting of its extracellular and transmembrane domains. 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 TGF-β-DNR 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. Detailed sequences of TGF-β-DNR and the armed DLL3 CARs are presented in Sequence IDs 525-529.

[0191] Sequence ID 525 (T3-P2A-TGF-β-DNR amino acid sequence) TIFF0007894963000158.tif70146 Sequence ID 526 (TGF-β-DNR-P2A-T3 amino acid sequence) TIFF0007894963000159.tif70147 Sequence ID 527 (T3-T2A-TGF-β-DNR amino acid sequence) TIFF0007894963000160.tif70147 Sequence ID 528 (TGF-β-DNR-T2A-T3 amino acid sequence) TIFF0007894963000161.tif70147 Sequence ID 529 (TGF-β-DNR amino acid sequence) TIFF0007894963000162.tif20146

[0192] All constructs were packaged in lentiviruses based on second-generation lentiviral systems. Next, the lentiviruses were transduced into primary T cells isolated from healthy donor PBMCs. Four days after transduction, the positivity rates for sdAb and TGF-β-DNR were detected by FACS (Figure 13B). The results showed that the sdAb positivity rates were equivalent for T3-P2A-TGF-β-DNR, T3-T2A-TGF-β-DNR, TGF-β-DNR-P2A-T3, and TGF-β-DNR-T2A-T3 CAR-T cells. However, the positivity rate for TGF-β-DNR was higher in the T3-P2A-TGF-β-DNR and T3-T2A-TGF-β-DNR constructs than in the TGF-β-DNR-P2A-T3 and TGF-β-DNR-T2A-T3 constructs. These results indicate that TGF-β-DNR expression is higher when it is conjugated to the C-terminus of CAR.

[0193] In vitro cytotoxicity assay Next, the cytotoxicity of these CAR-T cells was evaluated by LDH or IFN-γ release assays. Five days after transduction, CAR-T cells were adjusted to the same sdAb positivity using untransduced T cells (UnT). Then, CAR-T cells or UnT cells were co-incubated with SHP77 for 48 hours in the presence of 5 ng / mL of TGF-β, 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 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 cells showed higher levels of IFN-γ secretion than TGF-β-DNR-P2A-T3 and TGF-β-DNR-T2A-T3 CAR-T cells. In summary, these results indicate 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 effect of TGF-β on CAR-T cells, a long-term stimulation assay was performed. Specifically, T3-P2A-TGF-β-DNR and T3 CAR-T cells were repeatedly stimulated with SHP77 cells every 3 days, either in the presence or absence of 5 ng / mL of TGF-β. At the end of each stimulation cycle, the percentage of T cells in the total viable cells was analyzed by FACS, and the expansion and proliferation of CAR-T cells were calculated. As shown in Figures 13E and 13F, the persistence and expansion of T3 CAR-T cells were inhibited by TGF-β. In contrast, the persistence and expansion of T3-P2A-TGF-β-DNR cells were well maintained even in the presence of TGF-β. After two stimulation cycles with SHP77 cells, T cell depletion markers were analyzed by FACS. As shown in Figure 13G, treatment with TGF-β upregulated the expression of depletion markers in T3 cells, but not in T3-P2A-TGF-β-DNR CAR-T cells. In summary, these results indicate that TGF-β-DNR protects DLL3 CAR-T cells from TGF-β inhibition. Given that TGF-β expression levels are normally elevated in the solid tumor microenvironment, 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 trial To further investigate 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. Specifically, 1 × 10⁻⁶ 7 Individual SHP77 cells were subcutaneously transplanted into NCG mice. After 7-10 days, the tumor volume was 100-200 mm². 3 When it reaches 2.5 × 10 5100 CAR-T cells were intravenously injected into mice. Tumor volume was then measured twice weekly, and the percentage of CAR-T cells in peripheral blood was measured once weekly. As shown in Figure 13H, at suboptimal doses, T3-T2A-TGF-β-DNR CAR-T cells strongly suppressed tumor growth, while parental T3 CAR-T cells did not. As shown in Figure 13I, the percentage of T3-T2A-TGF-β-DNR CAR-T cells in peripheral blood leukocytes was higher than that of T3 cells. In summary, these results indicate that TGF-β-DNR enhanced the antitumor efficacy of DLL3 CAR-T cells in vivo.

[0196] Sequence information SEQUENCE LISTING <110> LEGEND BIOTECH IRELAND LIMITED <120> ANTI-DLL3 CHIMERIC ANTIGEN RECEPTORS AND USES THEREOF <150> PCT / CN2020 / 090587 <151> 2020-05-15 <150> PCT / CN2019 / 096360 <151> 2019-07-17 <160> 529 <170> EntIn version 3.3 <210> 1 <211> 10 <212> PRT <213> Artificial <220> <223> AS63930 CDR1 <400> 1 Gly Tyr Thr Tyr Ser Gly Asn Tyr Met Ala 1 5 10 <210> 2 <211> 10 <212> PRT <213> Artificial <220> <223> AS63932 CDR1 <400> 2 Gly Tyr Thr Tyr Gly Ser Thr Phe Met Gly 1 5 10 <210> 3 <211> 10 <212> PRT <213> Artificial <220> <223> AS63951 CDR1 <400> 3 Arg Asp Ile Tyr Gly Asn Asn Cys Met Ala 1 5 10 <210> 4 <211> 10 <212> PRT <213> Artificial <220> <223> AS63984 CDR1 <400> 4 Gly Tyr Thr Tyr Ser Ser Asn Phe Met Gly 1 5 10 <210> 5 <211> 7 <212> PRT <213> Artificial <220> <223> AS63987 CDR1 <400> 5 Gly Tyr Arg Asn Cys Met Ala 1 5 <210> 6 <211> 10 <212> PRT <213> Artificial <220> <223> AS63997 CDR1 <400> 6 Phe Ser Gly Tyr Gly Val Ser Thr Met Ala 1 5 10 <210> 7 <211> 10 <212> PRT <213> Artificial <220> <223> AS64047 CDR1 <400> 7 Gln Tyr Val Tyr Arg Trp Asp Leu Met Gly 1 5 10 <210> 8 <211> 10 <212> PRT <213> Artificial <220> <223> AS64052 CDR1 <400> 8 Gly Tyr Thr Tyr Arg Ser Asn Phe Met Gly 1 5 10 <210> 9 <211> 11 <212> PRT <213> Artificial <220> <223> AS64062 CDR1 <400> 9 Arg Ser Pro Tyr Ser Ser Ser Arg Cys Met Gly 1 5 10 <210> 10 <211> 10 <212> PRT <213> Artificial <220> <223> AS64072 CDR1 <400> 10 Gly Tyr Ser Tyr Tyr Ile Asn Leu Met Ala 1 5 10 <210> 11 <211> 10 <212> PRT <213> Artificial <220> <223> AS64097 CDR1 <400> 11 Gly Tyr Thr Tyr Ser Arg Asn Cys Met Gly 1 5 10 <210> 12 <211> 10 <212> PRT <213> Artificial <220> <223> AS64114 CDR1 <400> 12 Gly Asn Thr Tyr Ser Thr Asn Tyr Met Gly 1 5 10 <210> 13 <211> 10 <212> PRT <213> Artificial <220> <223> AS64123 CDR1 <400> 13 Gly Tyr Thr Tyr Thr Ser Asn Trp Leu Gly 1 5 10 <210> 14 <211> 10 <212> PRT <213> Artificial <220> <223> AS64130 CDR1 <400> 14 Gly Tyr Thr Tyr Arg Ser Asn Phe Met Gly 1 5 10 <210> 15 <211> 10 <212> PRT <213> Artificial <220> <223> AS64137 CDR1 <400> 15 Gly Ser Thr Tyr Ser Thr Asn Phe Met Gly 1 5 10 <210> 16 <211> 10 <212> PRT <213> Artificial <220> <223> AS64142 CDR1 <400> 16 Gly Phe Thr Phe Asp Arg Asn Ala Met Arg 1 5 10 <210> 17 <211> 10 <212> PRT <213> Artificial <220> <223> AS64154 CDR1 <400> 17 Gly Tyr Thr Tyr Arg Tyr Leu Tyr Met Gly 1 5 10 <210> 18 <211> 10 <212> PRT <213> Artificial <220> <223> AS64160 CDR1 <400> 18 Val Tyr Thr Ser Ser Ser Tyr Cys Met Gly 1 5 10 <210> 19 <211> 10 <212> PRT <213> Artificial <220> <223> AS64228 CDR1 <400> 19 Gly Val Ser Tyr Asn Arg Cys Ser Met Gly 1 5 10 <210> 20 <211> 10 <212> PRT <213> Artificial <220> <223> AS64300 CDR1 <400> 20 Gly Asp Ile Tyr Asn Leu Met Ser Met Ala 1 5 10 <210> 21 <211> 10 <212> PRT <213> Artificial <220> <223> AS64380 CDR1 <400> 21 Gly Asn Thr Tyr Ser Ser Asn Tyr Met Gly 1 5 10 <210> 22 <211> 10 <212> PRT <213> Artificial <220> <223> AS64395 CDR1 <400> 22 Gly Ser Thr Tyr Ser Thr Asn Phe Met Gly 1 5 10 <210> 23 <211> 10 <212> PRT <213> Artificial <220> <223> AS64443 CDR1 <400> 23 Gly Tyr Thr Asp Ser Ser Val Tyr Ile Gly 1 5 10 <210> 24 <211> 10 <212> PRT <213> Artificial <220> <223> AS64511 CDR1 <400> 24 Arg Ala Thr Tyr Ser Thr Asn Tyr Ile Ser 1 5 10 <210> 25 <211> 10 <212> PRT <213> Artificial <220> <223> AS64536 CDR1 <400> 25 Arg Tyr Thr Asp Asn Phe Val Tyr Met Gly 1 5 10 <210> 26 <211> 10 <212> PRT <213> Artificial <220> <223> AS64597 CDR1 <400> 26 Gly Tyr Thr Tyr Arg Val Asn Phe Met Gly 1 5 10 <210> 27 <211> 9 <212> PRT <213> Artificial <220> <223> AS64617 CDR1 <400> 27 Gly Tyr Thr Asp Arg Cys Ser Met Ala 1 5 <210> 28 <211> 12 <212> PRT <213> Artificial <220> <223> AS64634 CDR1 <400> 28 Gly Tyr Ser Phe Arg Gly Asp Phe Met Cys Met Gly 1 5 10 <210> 29 <211> 10 <212> PRT <213> Artificial <220> <223> AS69498 CDR1 <400> 29 Gly Asn Thr Tyr Ser Ser Asn Tyr Met Ala 1 5 10 <210> 30 <211> 10 <212> PRT <213> Artificial <220> <223> AS69500 CDR1 <400> 30 Arg Tyr Thr Tyr Ser Ser Ala Cys 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Asp Thr Thr Tyr Ile Gly 1 5 10 <210> 47 <211> 10 <212> PRT <213> Artificial <220> <223> AS64057 CDR1 <400> 47 Gly Phe Thr Phe Asp Arg Asn Ala Met Arg 1 5 10 <210> 48 <211> 10 <212> PRT <213> Artificial <220> <223> AS64060 CDR1 <400> 48 Gly Ser Thr Tyr Cys Thr Tyr Arg Met Ser 1 5 10 <210> 49 <211> 10 <212> PRT <213> Artificial <220> <223> AS64071 CDR1 <400> 49 Gly Asn Thr Tyr Arg Leu Asn Ser Met Gly 1 5 10 <210> 50 <211> 10 <212> PRT <213> Artificial <220> <223> AS64093 CDR1 <400> 50 Arg Tyr Ile Tyr Gly Asn Asn Cys Met Ala 1 5 10 <210> 51 <211> 10 <212> PRT <213> Artificial <220> <223> AS64118 CDR1 <400> 51 Gly Tyr Thr Tyr Ser Ala Cys Arg Met Ala 1 5 10 <210> 52 <211> 10 <212> PRT <213> Artificial <220> <223> AS64120 CDR1 <400> 52 Arg Tyr Ile Tyr Gly Asn Asn Cys Met Ala 1 5 10 <210> 53 <211> 10 <212> PRT <213> Artificial <220> <223> AS64124 CDR1 <400> 53 Thr Tyr Thr Pro Ser Asn Asn Tyr Met Gly 1 5 10 <210> 54 <211> 10 <212> PRT <213> Artificial <220> <223> AS64135 CDR1 <400> 54 Thr Ser Thr Tyr Cys Arg Tyr Tyr Met Arg 1 5 10 <210> 55 <211> 10 <212> PRT <213> Artificial <220> <223> AS64163 CDR1 <400> 55 Gly Tyr Arg Tyr Arg Trp Asn Cys Met Ala 1 5 10 <210> 56 <211> 10 <212> PRT <213> Artificial <220> <223> AS64182 CDR1 <400> 56 Gly Gln Thr Ser Arg Tyr Leu Tyr Met Gly 1 5 10 <210> 57 <211> 10 <212> PRT <213> Artificial <220> <223> AS64183 CDR1 <400> 57 Gly His Thr Tyr Ser Ala Asn Cys Met Ala 1 5 10 <210> 58 <211> 10 <212> PRT <213> Artificial <220> <223> AS64207 CDR1 <400> 58 Gly Tyr Thr Tyr Ser Ser Asn Phe Met Gly 1 5 10 <210> 59 <211> 10 <212> PRT <213> Artificial <220> <223> AS64276 CDR1 <400> 59 Gly Tyr Thr Gly Ser Ser Arg Cys Met Ala 1 5 10 <210> 60 <211> 10 <212> PRT <213> Artificial <220> <223> AS64336 CDR1 <400> 60 Gly Arg Thr Tyr Ser Ser Cys Ser Met Gly 1 5 10 <210> 61 <211> 7 <212> PRT <213> Artificial <220> <223> AS64346 CDR1 <400> 61 Gly Tyr Thr Tyr Phe Met Gly 1 5 <210> 62 <211> 10 <212> PRT <213> Artificial <220> <223> AS64420 CDR1 <400> 62 Gly Asp Thr Ser Arg Ser Val Trp Met Gly 1 5 10 <210> 63 <211> 10 <212> PRT <213> Artificial <220> <223> AS64473 CDR1 <400> 63 Gly Tyr Thr Tyr Arg Tyr Leu Tyr Met Ala 1 5 10 <210> 64 <211> 10 <212> PRT <213> Artificial <220> <223> AS64475 CDR1 <400> 64 Gly Tyr Thr Trp Ser Arg Asn Trp Met Gly 1 5 10 <210> 65 <211> 10 <212> PRT <213> Artificial <220> <223> AS64513 CDR1 <400> 65 Asp Tyr Pro Tyr Ile Asp Asn Cys Met Gly 1 5 10 <210> 66 <211> 10 <212> PRT <213> Artificial <220> <223> AS64562 CDR1 <400> 66 Gly Tyr Thr Ala Arg Arg Asp Phe Met Ala 1 5 10 <210> 67 <211> 10 <212> PRT <213> Artificial <220> <223> AS64583 CDR1 <400> 67 Gly Phe Thr Ile Ala Val Tyr Thr Met Gly 1 5 10 <210> 68 <211> 10 <212> PRT <213> Artificial <220> <223> AS64594 CDR1 <400> 68 Gly Tyr Thr Tyr Asn Ser Asn Tyr Met Gly 1 5 10 <210> 69 <211> 10 <212> PRT <213> Artificial <220> <223> AS64605 CDR1 <400> 69 Arg Tyr Pro Tyr Ser Ser Ile Cys Met Gly 1 5 10 <210> 70 <211> 10 <212> PRT <213> Artificial <220> <223> AS64606 CDR1 <400> 70 Gly Tyr Thr Ser Arg Ser Asn Tyr Met Gly 1 5 10 <210> 71 <211> 10 <212> PRT <213> Artificial <220> <223> AS68121 CDR1 <400> 71 Gly Tyr Thr Tyr Ser Arg Asn Cys Met Gly 1 5 10 <210> 72 <211> 10 <212> PRT <213> Artificial <220> <223> AS68170 CDR1 <400> 72 Gly Tyr Thr Tyr Arg Ser Asn Cys Met Gly 1 5 10 <210> 73 <211> 10 <212> PRT <213> Artificial <220> <223> AS63964 CDR1 <400> 73 Gly Tyr Thr Tyr Ser Tyr Asn Asn Met Gly 1 5 10 <210> 74 <211> 8 <212> PRT <213> Artificial <220> <223> AS64116 CDR1 <400> 74 Gly Tyr Ile Tyr Ser Cys Val Gly 1 5 <210> 75 <211> 13 <212> PRT <213> Artificial <220> <223> AS68270 CDR1 <400> 75 Gly Tyr Pro Ser Ser Thr Tyr Tyr Met Leu Ser Met Ala 1 5 10 <210> 76 <211> 10 <212> PRT <213> Artificial <220> <223> AS68320 CDR1 <400> 76 Gly Tyr Thr Tyr Asn Thr Asn Tyr Met Gly 1 5 10 <210> 77 <211> 10 <212> PRT <213> Artificial <220> <223> AS68351 CDR1 <400> 77 Gly Asp Thr Phe Arg Ala Tyr Tyr Met Asn 1 5 10 <210> 78 <211> 10 <212> PRT <213> Artificial <220> <223> AS75378 CDR1 <400> 78 Gly Asn Thr Arg Ser Thr Thr Tyr Met Gly 1 5 10 <210> 79 <211> 10 <212> PRT <213> Artificial <220> <223> AS75383 CDR1 <400> 79 Gly Tyr Thr Phe Ser Ser Tyr Cys Leu Gly 1 5 10 <210> 80 <211> 11 <212> PRT <213> Artificial <220> <223> AS75751 CDR1 <400> 80 Gly Tyr Phe Tyr Asn Thr Tyr Tyr Phe Met Gly 1 5 10 <210> 81 <211> 10 <212> PRT <213> Artificial <220> <223> AS76422 CDR1 <400> 81 Gly Tyr Thr Phe Ala Gly Asn Cys Leu Gly 1 5 10 <210> 82 <211> 20 <212> PRT <213> Artificial <220> <223> AS63930 CDR2 <400> 82 Val Val Tyr Asn Ile Asp Gly Gly Gly Arg Phe Thr Thr Tyr Ala Asp 1 5 10 15 Ser Val Lys Gly 20 <210> 83 <211> 17 <212> PRT <213> Artificial <220> <223> AS63932 CDR2 <400> 83 Val Ile Tyr Thr Gly Gly Gly Ser Thr Trp Tyr Ala Ser Ser Val Lys 1 5 10 15 Gly <210> 84 <211> 17 <212> PRT <213> Artificial <220> <223> AS63951 CDR2 <400> 84 Ser Ile Tyr Pro Ala Gly Gly Arg Pro Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <210> 85 <211> 17 <212> PRT <213> Artificial <220> <223> AS63984 CDR2 <400> 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 <211> 125 <212> PRT <213> Artificial <220>...

Claims

1. A chimeric antigen receptor (CAR) that specifically binds to DLL3, wherein the CAR comprises a first single-domain antibody (sdAb) moiety and a second sdAb moiety, The first sdAb portion includes CDR1 containing the amino acid sequence of SEQ ID NO: 6; CDR2 containing the amino acid sequence of SEQ ID NO: 87; and CDR3 containing the amino acid sequence of SEQ ID NO: 168; and The second sdAb portion includes CDR1 containing the amino acid sequence of SEQ ID NO: 21; CDR2 containing the amino acid sequence of SEQ ID NO: 102; and CDR3 containing the amino acid sequence of SEQ ID NO:

183. The aforementioned CAR.

2. The first sdAb portion includes the amino acid sequence of SEQ ID NO: 366, and the second sdAb portion includes the amino acid sequence of SEQ ID NO:

356. The CAR according to claim 1.

3. The CAR according to claim 1, wherein the CAR comprises a signal peptide, a DLL3 binding domain, a hinge domain, a transmembrane domain, and an intracellular signal transduction domain, from the N-terminus to the C-terminus, wherein the DLL3 binding domain comprises the first sdAb portion and the second sdAb portion.

4. The CAR according to claim 3, wherein the intracellular signaling domain includes the intracellular signaling domains of CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CD5, CD22, CD79a, CD79b, or CD66d.

5. The CAR according to claim 3, wherein the intracellular signaling domain further comprises an intracellular costimulatory sequence.

6. The CAR according to claim 5, wherein the intracellular co-stimulatory sequence includes a sequence of a co-stimulatory 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, the ligand of CD83, and combinations thereof.

7. The CAR according to claim 1, comprising an amino acid sequence having at least 95% sequence identity with respect to the amino acid sequence of SEQ ID NO:

520.

8. A nucleic acid molecule encoding CAR as described in claim 1.

9. The nucleic acid molecule according to claim 8, further comprising a polynucleotide sequence encoding a PD-1 dominant-negative receptor (PD-1 DNR) or a PD-1 chimeric switch receptor (PD-1 CSR).

10. The nucleic acid molecule according to claim 9, wherein the PD-1 DNR comprises an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 523; and the PD-1 CSR comprises an amino acid sequence having at least 95% sequence identity with SEQ ID NO:

524.

11. The nucleic acid molecule according to claim 9, wherein the polynucleotide sequence encoding the PD-1 DNR or the PD-1 CSR is linked to the polynucleotide sequence encoding the CAR via a polynucleotide sequence encoding a 2A self-cleaving peptide.

12. The nucleic acid molecule according to claim 11, wherein the 2A self-cleaving peptide is a T2A peptide or a P2A peptide.

13. The nucleic acid molecule according to claim 11, comprising, in the direction from 5' to 3', a polynucleotide sequence encoding the CAR, a polynucleotide sequence encoding the 2A self-cleaving peptide, and a polynucleotide sequence encoding the PD-1 DNR or the PD-1 CSR.

14. The nucleic acid molecule according to claim 9, which encodes a peptide having at least 95% sequence identity with sequence number 521 or 522.

15. An expression vector comprising the nucleic acid molecule described in claim 8.

16. Modified immune cells comprising the nucleic acid molecule described in claim 8.

17. A modified immune cell according to claim 16, selected from the group consisting of cytotoxic T cells, helper T cells, natural killer T cells, γδ T cells, and natural killer cells.

18. A pharmaceutical composition comprising the modified immune cells described in claim 16 and a physiologically acceptable excipient.

19. A drug for the treatment of cancer in a subject, comprising a therapeutically effective amount of the pharmaceutical composition described in claim 18, wherein the cancer is 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.

20. The agent according to claim 19, wherein the cancer is small cell lung cancer.

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