Immunoconjugates for NCAM1 and methods of use

Abstract

Provided herein are anti-NCAMl antibodies and anti-NCAMl immunoconjugates. Such anti-NCAMl antibodies and anti-NCAMl immunoconjugates can be used for treating cancer.

Description

Attorney Docket No. 22419-20010.40IMMUNOCONJUGATES FOR NCAM1 AND METHODS OF USECROSS REFERNCE TO RELATED APPLICATIONS

[0001] This application claims priority benefit U.S. Provisional Application Nos. 63 / 725,269, 63 / 737,311, 63 / 789,294, 63 / 812,732, 63 / 815,126, and 63 / 903,764, filed November 26, 2024, December 20, 2024, April 15, 2025, May 27, 2025, May 30, 2025, and October 22, 2025, respectively, the contents of each of which are hereby incorporated by reference in their entireties.REFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0002] The contents of the electronic sequence listing (224192001040SEQLIST.xml; Size: 46,268 bytes; and Date of Creation: November 21, 2025) is herein incorporated by reference in its entirety.FIELD OF INVENTION

[0003] The invention relates to anti-NCAMl antibodies and anti-NCAMl immunoconjugates, and methods of using the same for treating cancer.BACKGROUND

[0004] Neural cell adhesion molecule 1 (NCAM1), also called CD56, is a member of the immunoglobulin super family. NCAM1 is highly expressed in natural killer cells, T and B lymphocytes, and neural and mesenchymal stem cells. NCAM1 is involved in cell proliferation and expansion, cell adhesion, neurogenesis, neurite outgrowth, and cell migration. Several cancer cell types express NCAM1 including several solid tumors such as multiple myeloma, glioblastoma, sarcoma, small cell lung cancer (SCLC), and ovarian cancer.

[0005] Previous antibody-drug conjugate (ADC) therapies for NCAM1 have been largely unsuccessful. Lorvotuzumab mertansine is an ADC composed of an anti-CD56 humanized monoclonal antibody (hN901) conjugated to mertansine. Lorvotuzumab mertansine suffered from poor pharmacokinetic properties, an unstable linker technology, and a payload mechanism of action that was not well suited for tumors like SCLC. This is believed to be due, in part, to the high antigen-mediated clearance due to CD56 expression on a variety of normal tissues, in particular highly abundant NK cells, which as a result, restricts1MF-363628402Attorney Docket No. 22419-20010.40Lorvotuzumab mertansine’s exposure and thus activity in tumor killing. As such improved anti-NCAMl antibodies and ADCs are needed.BRIEF SUMMARY OF INVENTION

[0006] Provided herein are antibodies and immunoconjugates (e.g., ADCs) for NCAM1 that address the previous limitations of prior treatments. For example, ADCs for NCAM1 described herein have improved biophysical and pharmacokinetic properties including halflife, increased linker stability, and / or improved delivery of the drug to the tumor, and as such represent an improved therapeutic.

[0007] Described herein, in certain embodiments, are antibodies or antigen-binding fragments thereof that bind neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 9 or at least about 90% sequence identity to SEQ ID NO: 10.

[0008] Described herein, in certain embodiments, are antibodies or antigen-binding fragments thereof that bind neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising an amino acid sequence according to SEQ ID NO: 11; and b) a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 9 or at least about 90% sequence identity to SEQ ID NO: 10.

[0009] Described herein, in certain embodiments, are antibodies or antigen-binding fragments thereof that bind neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 12.

[0010] Described herein, in certain embodiments, are antibodies or antigen-binding fragments thereof that bind neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 7, at least about 99% sequence identity SEQ ID NO: 8, or an amino acid sequence according to SEQ ID NO: 11; and b) a light chain variable region (VL) comprising an amino acid sequence having at least about 90% sequence identity to the amino acid sequence of SEQ ID NO: 12, and wherein:(i) the VL comprises a substitution at amino acid position 79 (sequential) relative to the reference sequence of SEQ ID NO: 12;2MF-363628402Attorney Docket No. 22419-20010.40(ii) the VL comprises a substitution at amino acid position 82 (sequential) relative to the reference sequence of SEQ ID NO: 12;(iii) the VL comprises substitutions at amino acid positions 79 and 82 (sequential) relative to the reference sequence of SEQ ID NO: 12;(iv) the VL comprises a substitution at amino acid position 74 (Kabat) relative to the reference sequence of SEQ ID NO: 12;(v) the VL comprises a substitution at amino acid position 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12; or(vi) the VL comprises substitutions at amino acid positions 74 and 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12.

[0011] Described herein, in certain embodiments, are antibodies or antigen-binding fragments thereof that bind neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and b) a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); wherein the antibody or antigen-binding fragment is humanized.

[0012] In some embodiments according to any of the embodiments described above, the antibody or antigen-binding fragment thereof comprises: a) a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 9; b) a VH comprising an amino acid sequence according to SEQ ID NO: 8, and a VL comprising an amino acid sequence according to SEQ ID NO: 10; c) a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 10; d) a VH comprising an amino acid sequence according to SEQ ID NO: 8, and a VL comprising an amino acid sequence according to SEQ ID NO: 9;3MF-363628402Attorney Docket No. 22419-20010.40 e) a VH comprising an amino acid sequence according to SEQ ID NO: 11, and a VL comprising an amino acid sequence according to SEQ ID NO: 9; f) a VH comprising an amino acid sequence according to SEQ ID NO: 11, and a VL comprising an amino acid sequence according to SEQ ID NO: 10; g) a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 12; or h) a VH comprising an amino acid sequence according to SEQ ID NO: 8, and a VL comprising an amino acid sequence according to SEQ ID NO: 12.

[0013] Described herein, in certain embodiments, are antibodies or antigen-binding fragments thereof that bind neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and b) a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

[0014] In some embodiments according to any of the embodiments described above, the antibody or the antigen-binding fragment thereof is humanized.

[0015] Described herein, in certain embodiments, are immunoconjugates comprising: a) an antibody or antigen-binding fragment thereof that binds to neural cell adhesion molecule 1 (NCAM1), wherein the antibody or the antigen-binding fragment thereof comprises:(I) i) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and ii) a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE);4MF-363628402Attorney Docket No. 22419-20010.40(ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); or(II) i) a VH comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and ii) a VL comprising (i) a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6; and b) a topoisomerase I inhibitor conjugated to the antibody or the antigen-binding fragment thereof using a linker.

[0016] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Cehaloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and - [(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;5MF-363628402Attorney Docket No. 22419-20010.40R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)pO(CY2)q]tCY3;R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-C6haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci- Ce haloalkynyl)-X2;X1is — O— , — S(O)W— , — NH— , — O— (C=O)— , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ;X2is —OR9, —SR9, or — NHR9;R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually hydrogen or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6;R7is hydrogen, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker;R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)PO(CY2)q]tCY2 — X3;R9is hydrogen, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker, with the proviso that exactly one of R7and R9is the point of attachment to the linker; and each X3is individually — H, — OH, — SH, or — NH2.6MF-363628402Attorney Docket No. 22419-20010.40

[0017] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, -OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O-(Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or an unsubstituted Ci-Ce alkyl;R4is a substituted or an unsubstituted — (Ci-Ce alkyl)-X2;X1is — O— ;X2is —OR9;R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0;R7is hydrogen; andR9is the point of attachment to the linker.

[0018] In some embodiments according to any of the embodiments described above, n4is 1.

[0019] In some embodiments according to any of the embodiments described above, n4is 2.7MF-363628402Attorney Docket No. 22419-20010.40

[0020] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:R1is methyl;R2is fluoro;R3is hydrogen;R4is a substituted or an unsubstituted -(C2 alkyl)-X2; and n4is 2, and wherein the remaining variables are as described herein.

[0021] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor is represented by:wherein R9is the point of attachment to the linker.

[0022] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor is represented by:8MF-363628402Attorney Docket No. 22419-20010.40wherein R9is the point of attachment to the linker.

[0023] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor is represented by:wherein R9is the point of attachment to the linker.

[0024] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor and the linker, taken together, are represented by:9MF-363628402Attorney Docket No. 22419-20010.40

[0025] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor and the linker, taken together, are represented by:

[0026] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor and the linker, taken together, are represented by:10MF-363628402Attorney Docket No. 22419-20010.40

[0027] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor and the linker, taken together, are represented by:

[0028] In some embodiments according to any of the embodiments described above, the topoisomerase I inhibitor and the linker, taken together, are represented by:11MF-363628402Attorney Docket No. 22419-20010.40

[0029] Described herein, in certain embodiments, are immunoconjugates having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising:(I) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); or(II) a VH comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and a VL comprising (i) a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L1is12MF-363628402Attorney Docket No. 22419-20010.40Z1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl;L3is -(CH2)n1-C(=O)- or -(CH2CH2O)n1-(CH2)n1C(=O)-; each n1is individually an integer from 0 to 10;L4is a tetrapeptide residue;L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent,D is a drug moiety; and n is an integer from 1 to 10.13MF-363628402Attorney Docket No. 22419-20010.40

[0030] In some embodiments according to any of the embodiments described above, the drug moiety is represented by Formula (II) having the structure:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and - [(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3;R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-C6haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci- Ce haloalkynyl)-X2;X1is — O— , — S(O)W— , — NH— , — O— (C=O)— , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ;X2is —OR9, —SR9, or — NHR9;14MF-363628402Attorney Docket No. 22419-20010.40R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-C6alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2) PO(CY2)q]tCY.3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually hydrogen or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6;R7is hydrogen, —COR8, — CO2R8, — (CO)— NHR8, L4, L5, L6, or L7;R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)PO(CY2)q]tCY2 — X3;R9is hydrogen, — COR8, — CO2R8, — (CO) — NHR8, L4, L5, L6, or L7, with the proviso that exactly one of R7and R9is L4, L5, L6, or L7; and each X3is individually — H, — OH, — SH, or — NH2.

[0031] In some embodiments according to any of the embodiments described above, the drug moiety of Formula (II) is represented by:wherein:15MF-363628402Attorney Docket No. 22419-20010.40R1and R2are each individually selected from the group consisting of hydrogen, halogen, -OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O-(Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or an unsubstituted Ci-Ce alkyl;R4is a substituted or an unsubstituted — (Ci-Ce alkyl)-X2;X1is — O— ;X2is —OR9;R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0;R7is hydrogen; andR9is L4, L5, L6, or L7;

[0032] In some embodiments according to any of the embodiments described above, n4is 1.

[0033] In some embodiments according to any of the embodiments described above, n4is 2.

[0034] In some embodiments according to any of the embodiments described above, the drug moiety of Formula (II) is represented by:R1is methyl;R2is fluoro;R3is hydrogen;R4is a substituted or an unsubstituted -(C2 alkyl)-X2; and n4is 2,16MF-363628402Attorney Docket No. 22419-20010.40 and wherein the remaining variables are as described herein.

[0035] In some embodiments according to any of the embodiments described above,L2is absent; n1is the integer 2 or 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; andL7is absent, and wherein the remaining variables are as described herein.

[0036] In some embodiments according to any of the embodiments described above, the drug moiety of Formula (II) is represented by:wherein R9is L4, L5, L6, or L7.

[0037] In some embodiments according to any of the embodiments described above, the drug moiety of Formula (II) is represented by:wherein R9is L4, L5, L6, or L7.

[0038] In some embodiments according to any of the embodiments described above, the drug moiety of Formula (II) is represented by:17MF-363628402Attorney Docket No. 22419-20010.40

[0039] In some embodiments according to any of the embodiments described above, Formula(I) is:wherein n is as described herein.

[0040] In some embodiments according to any of the embodiments described above, Formula(I) is:18MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.

[0041] In some embodiments according to any of the embodiments described above, Formula(I) is:19MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.

[0042] In some embodiments according to any of the embodiments described above, Formula(I) is represented by:wherein n is as described herein.

[0043] In some embodiments according to any of the embodiments described above, Formula(I) is represented by:20MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.

[0044] Described herein, in certain embodiments, are immunoconjugates having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising:(I) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to (SFGMH) SEQ ID NO: 1;(ii) a CDR2 having an amino acid sequence according to (YISSGSFTIYYADSVKG) SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to (MRKGYAMDY) SEQ ID NO: 3; and a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to (RSSQIIIHSDGNTYLE) SEQ ID NO: 4; (ii) a CDR2 having an amino acid sequence according to (KVSNRFS) SEQ ID NO: 5; and (iii) a CDR3 having an amino acid sequence according to (FQGSHVPHT) SEQ ID NO: 6; or21MF-363628402Attorney Docket No. 22419-20010.40(II) a VH comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and a VL comprising (i) a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L1isZ1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl;L3is -(CH2)n1-C(=O)- or -(CH2CH2O)n1-(CH2)n1C(=O)-; each n1is individually an integer from 0 to 10;L4is a tetrapeptide residue;L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent,22MF-363628402Attorney Docket No. 22419-20010.40wherein D has the structure:and n is an integer from 1 to 10.

[0045] In some embodiments according to any of the embodiments described above, n is an integer from 4 to 8.

[0046] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 7 or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 8.

[0047] In some embodiments according to any of the embodiments described above, the VL comprises an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 9 or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 10.23MF-363628402Attorney Docket No. 22419-20010.40

[0048] In some embodiments according to any of the embodiments described above, the antibody or the antigen-binding fragment thereof is humanized.

[0049] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 7 or SEQ ID NO: 8.

[0050] In some embodiments according to any of the embodiments described above, the VL comprises an amino acid sequence according to SEQ ID NO: 9 or SEQ ID NO: 10.

[0051] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

[0052] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

[0053] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

[0054] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

[0055] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

[0056] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

[0057] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 12.

[0058] In some embodiments according to any of the embodiments described above, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 12.

[0059] In some embodiments according to any of the embodiments described above, the antibody comprises a Fc domain comprising one or more modifications that reduces binding to an Fc receptor, reduces binding to complement component Clq, or both binding to an Fc receptor and reduces binding to complement component Clq.24MF-363628402Attorney Docket No. 22419-20010.40

[0060] In some embodiments according to any of the embodiments described above, the antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0061] In some embodiments according to any of the embodiments described above, the antibody or the antigen-binding fragment thereof binds to human NCAM1 with an affinity of 10’7M to 10’10M.

[0062] Described herein, in certain embodiments, are immunoconjugates having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a heavy chain variable region (VH) comprising the amino acid sequence according to SEQ ID NO: 7; and a light chain variable region (VL) comprising the amino acid sequence according to SEQ ID NO: 10;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L2is absent;L3is -(CH2)n1-C(=O)-; n1is the integer 2;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; and L6is absent;L7is absent;D has the structure:25MF-363628402Attorney Docket No. 22419-20010.40and n is 8.

[0063] Described herein, in certain embodiments, are immunoconjugates having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a heavy chain variable region (VH) comprising the amino acid sequence according to SEQ ID NO: 7; and a light chain variable region (VL) comprising the amino acid sequence according to SEQ ID NO: 10;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L2is absent;L3is -(CH2)n1-C(=O)-; n1is the integer 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; and L6is absent;26MF-363628402Attorney Docket No. 22419-20010.40L7is absent;D has the structure:and n is 8.

[0064] Described herein, in certain embodiments, are polynucleotides encoding any of the antibodies or the antigen-binding fragments thereof described herein or the polypeptide components of any of the immunoconjugates described herein.

[0065] Described herein, in certain embodiments, are expression vectors comprising any of the polynucleotides described herein.

[0066] Described herein, in certain embodiments, are cells comprising any of the expression vectors described herein.

[0067] Described herein, in certain embodiments, are cells capable of producing any of the antibodies or the antigen-binding fragments thereof described herein or the polypeptide components of any of the immunoconjugates described herein.

[0068] Described herein, in certain embodiments, are pharmaceutical compositions comprising any of the antibodies or the antigen-binding fragments thereof described herein or any of the immunoconjugates described herein, and a pharmaceutically acceptable excipient, salt, carrier, or diluent.

[0069] Described herein, in certain embodiments, are pharmaceutical compositions for use in treating cancer in a subject.

[0070] Described herein, in certain embodiments, are methods of treating cancer comprising administering a therapeutically effective amount of any of the antibodies or the antigenbinding fragments thereof described herein or any of the immunoconjugates described herein to a subject in need thereof.27MF-363628402Attorney Docket No. 22419-20010.40

[0071] In some embodiments according to any of the embodiments described above, the method further comprises administering an additional anti-cancer therapy selected from the group consisting of surgery, chemotherapy, radiotherapy, and immunotherapy.

[0072] In some embodiments according to any of the embodiments described above, the method further comprises administering to the subject an immune-modulator, activated lymphocyte cell, kinase inhibitor, or a chemotherapeutic agent.

[0073] In some embodiments according to any of the embodiments described above, the cancer is a solid cancer.

[0074] In some embodiments according to any of the embodiments described above, the cancer is a neuroendocrine cancer.

[0075] In some embodiments according to any of the embodiments described above, the method further comprises the cancer is multiple myeloma, glioblastoma, sarcoma, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, or small intestinal neuroendocrine tumor.

[0076] In some embodiments according to any of the embodiments described above, the cancer is NCAM1 positive.

[0077] Described herein, in certain embodiments, are methods of diagnosing or prognosing cancer in a subject, the method comprising determining the expression level of NCAM1 in a biological samples using any of the antibodies or the antigen-binding fragments thereof described herein or any of the immunoconjugates described herein.

[0078] Described herein, in certain embodiments, are uses of an effective amount of any of the antibodies or the antigen-binding fragments thereof described herein or any of the immunoconjugates described herein for treating cancer.

[0079] In some embodiments according to any of the embodiments described above, the cancer is a solid cancer.

[0080] In some embodiments according to any of the embodiments described above, the cancer is a neuroendocrine cancer.

[0081] In some embodiments according to any of the embodiments described above, the cancer is multiple myeloma, glioblastoma, sarcoma, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, or small intestinal neuroendocrine tumor.28MF-363628402Attorney Docket No. 22419-20010.40

[0082] Described herein, in certain embodiments, are kits comprising any of the antibodies or the antigen-binding fragments thereof described herein or any of the immunoconjugates described herein and instructions for use.

[0083] Described herein, in certain embodiments, are compositions comprising a plurality of any of the immunoconjugates described herein, wherein the average n of the plurality of the immunoconjugates is a number from 1 to 10.BRIEF DESCRIPTION OF THE DRAWINGS

[0084] FIG. 1 depicts thermal stability data of anti-NCAMl antibodies described herein.

[0085] FIGs. 2A-2B depict binding data of anti-NCAMl antibodies described herein.

[0086] FIG. 3 depicts the mouse pharmacokinetics of anti-NCAMl antibodies described herein.

[0087] FIGs. 4A-4B depict NCAM1 mRNA expression data across multiple cancer types (FIG. 4A) and neuroendocrine cancers (FIG. 4B).

[0088] FIGs. 5A-5B depict tumor volume reduction in NCI-H526 small cell lung cancer (SCLC) cell line-derived xenografts treated with anti-NCAMl ADCs described herein.

[0089] FIG. 5C depicts tumor volume reduction in cell line-derived xenograft models including in NCI-H810 (a model for large cell neuroendocrine carcinoma; LCNEC), NCI- H660 (a model for neuroendocrine prostate cancer; NEPC), QGP-1 (a model for pancreatic neuroendocrine tumor; PNET), and SK-N-FI (a model for neuroblastoma; NBL) using Antibody 3 ADC.

[0090] FIG. 6A depicts the cynomolgus monkey pharmacokinetics of anti-NCAMl ADCs described herein.

[0091] FIG. 6B depicts tumor volume reduction in NCI-H526 small cell lung cancer (SCLC) xenografts after treatment with Antibody 3 ADC, Lorvotuzumab ADC, or Lorvotuzumab mertansine. The ADC dose used in FIG. 6B was half of the ADC dose used in FIG. 5B.

[0092] FIG. 7A depicts a graph of MDCK II cell permeability of payload with Formula (II) as compared to exatecan, YL0014, tirumotecan, rezetecan, A- 1743332, and deruxtecan (DXd).

[0093] FIG. 7B depicts a graph showing topoisomerase I (TOPl)-mediated DNA relaxation activity of payload with Formula (II) compared to deruxtecan (DXd).

[0094] FIG. 7C depicts a schema and data relating to determination of in vitro bystander activity of payload with Formula (II) and deruxtecan (DXd) in a co-culture model using NCL H446 cells engineered to express either TROP2 or luciferase.29MF-363628402Attorney Docket No. 22419-20010.40

[0095] FIG. 8 depicts tumor volume reduction in a patient-derived xenograft model of Merkel cell carcinoma after treatment with Antibody 3 ADC.DETAILED DESCRIPTION

[0096] To facilitate an understanding of the present disclosure, a number of terms and phrases are defined below.

[0097] As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the content dictates otherwise. Thus, for example, reference to “an antibody” optionally includes a combination of two or more such molecules and the like.

[0098] The term “about,” as used herein, refers to the usual error range for the respective value readily known to the skilled person in this technical field; for example, ± 20%, ± 10%, or ± 5%, are within the intended meaning of the recited value.

[0099] As used herein, the term “antibody” means an isolated or recombinant binding agent that comprises the necessary variable region sequences to specifically bind an antigenic epitope. Therefore, an “antibody” as used herein is any form of an antibody of any class or subclass or fragment thereof that exhibits the desired biological activity, e.g., binding a specific target antigen. Thus, it is used in the broadest sense and specifically covers a monoclonal antibody (including full-length monoclonal antibodies), human antibodies, chimeric antibodies, nanobodies (Nbs), minibodies, small immunoproteins (SIPs), variable new antigen receptors (VNARs), diabodies, multispecific antibodies (e.g., bispecific antibodies), antibody fragments including but not limited to scFv, Fab, VHH, and the like so long as they exhibit the desired biological activity. In general, antibodies are multimeric proteins that contain four polypeptide chains. Two of the polypeptide chains are called immunoglobulin heavy chains (H chains), and two of the polypeptide chains are called immunoglobulin light chains (L chains). The immunoglobulin heavy and light chains are connected by an interchain disulfide bond. The immunoglobulin heavy chains are connected by interchain disulfide bonds. A light chain consists of one variable region (VL) and one constant region (CL). The heavy chain consists of one variable region (VH) and at least three constant regions (CHI, CH2 and CH3). The variable regions determine the binding specificity of the antibody. Each variable region contains three hypervariable regions known as complementarity determining regions (CDRs) flanked by four relatively conserved regions known as framework regions (FRs). The extent of the FRs and CDRs has been defined (Kabat, E.A., et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No. 91-3242; and Chothia, 30MF-363628402Attorney Docket No. 22419-20010.40C. et al. (1987) J. Mol. Biol. 196:901-917). The three CDRs in each variable region (e.g., light chain variable region or heavy chain variable region, with six CDRs total in a typical antibody format), referred to as CDR1, CDR2, and CDR3, collectively contribute to antibody binding specificity. Naturally occurring antibodies have been used as starting material for engineered antibodies, such as chimeric antibodies and humanized antibodies. Examples of antibodies that have been modified or engineered include chimeric antibodies, humanized antibodies, and multispecific antibodies (e.g., bispecific antibodies). An example of a chemically conjugated antibody is an antibody conjugated to a toxin moiety.

[0100] “Antibody fragments” comprise a portion of an intact antibody, for example, the antigen-binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab’, F(ab’)2, and Fv fragments; diabodies; linear antibodies (e.g., Zapata et al., Protein Eng. 8(10): 1057-1062 (1995)); single-chain antibody molecules (e.g., scFv); and multispecific antibodies formed from antibody fragments. Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen-binding site, and a residual “Fc” fragment, a designation reflecting the ability to crystallize readily. Pepsin treatment yields an F(ab’)2 fragment with two antigen combining sites and is still capable of cross-linking antigen.

[0101] As used herein, the term “anti-NCAMl antibody”, “antibody that binds NCAM1,” “antibody or antigen-binding fragment thereof that binds NCAM1,” “antibody that targets NCAM1,” or “NCAM1 -targeting antibody,” with respect to an antibody, refers to an antibody or antigen-binding fragment that binds preferentially to NCAM1.

[0102] As used herein, “V-region” refers to an antibody variable region domain comprising the segments of Framework 1, CDR1, Framework 2, CDR2, and Framework 3, including CDR3 and Framework 4. The heavy chain V-region, VH, is a consequence of the rearrangement of a V-gene (HV), a D-gene (HD), and a J-gene (HJ), in what is termed V(D)J recombination during B-cell differentiation. The light chain V-region, VE, is a consequence of the rearrangement of a V-gene (LV) and a J-gene (LJ).

[0103] As used herein, “complementarity-determining region (CDR)” refers to the three hypervariable regions (HVRs) in each chain that interrupt the four “framework” regions established by the light and heavy chain variable regions. The CDRs are the primary contributors to binding to an epitope of an antigen. The CDRs of each chain are referred to as CDR1, CDR2, and CDR3, numbered sequentially starting from the N-terminus, and are also identified by the chain in which the CDR is located. Thus, a VHCDR3 (CDR3) is in the variable domain of the heavy chain of the antibody in which it is found, whereas a VL CDR3 31MF-363628402Attorney Docket No. 22419-20010.40(CDR3) is the CDR3 from the variable domain of the light chain of the antibody in which it is located. The term “CDR” is used interchangeably with “HVR” when referring to CDR sequences.

[0104] The amino acid sequences of the CDRs and framework regions can be determined using various well-known definitions in the art, e.g., Kabat, Chothia, international ImMunoGeneTics database (IMGT), and AbM (see, e.g., Chothia & Lesk, 1987, Canonical structures for the hypervariable regions of immunoglobulins. J. Mol. Biol. 196, 901-917; Chothia C. et al., 1989, Conformations of immunoglobulin hypervariable regions. Nature 342, 877-883; Chothia C. et al., 1992, Structural repertoire of the human VH segments J. Mol. Biol. 227, 799-817; Al-Lazikani et al., J. Mol. Biol. 1997, 273(4)). Definitions of antigen combining sites are also described in the following: Ruiz et al., IMGT, the international ImMunoGeneTics database. Nucleic Acids Res., 28, 219-221 (2000); and Lefranc, M.-P. IMGT, the international ImMunoGeneTics database. Nucleic Acids Res. Jan l;29(l):207-9 (2001); MacCallum et al., Antibody-antigen interactions: Contact analysis and binding site topography, J. Mol. Biol., 262 (5), 732-745 (1996); and Martin et al, Proc. Natl Acad. Sci. USA, 86, 9268-9272 (1989); Martin et al., Methods Enzymol., 203, 121-153, (1991); Pedersen et al., Immunomethods, 1, 126, (1992); and Rees et al., In Sternberg M.J.E. (ed.), Protein Structure Prediction. Oxford University Press, Oxford, 141-172 1996).Reference to CDRs as determined by Kabat numbering is based, for example, on Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institute of Health, Bethesda, MD (1991)). Chothia CDRs are determined as defined by Chothia (see, e.g., Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)).

[0105] Table A provides the positions of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 as identified by the Kabat and Chothia schemes. For CDR-H1, residue numbering is provided using both the Kabat and Chothia numbering schemes. CDRs may be assigned, for example, using antibody numbering software, such as Abnum, available at www.bioinf.org.uk / abs / abnum / , and described in Abhinandan and Martin (2008) Immunology, 45:3832-3839, incorporated by reference in its entirety.Table A. Residues in CDRs according to Kabat and Chothia numbering schemes.32MF-363628402Attorney Docket No. 22419-20010.40* The C -terminus of CDR-H1, when numbered using the Kabat numbering convention, varies between H32 and H34, depending on the length of the CDR.

[0106] The “EU numbering scheme” is generally used when referring to a residue in an antibody heavy chain constant region (e.g., as reported in Kabat et al., supra). Unless stated otherwise, the EU numbering scheme is used to refer to residues in antibody heavy chain constant regions described herein.

[0107] An “Fc region” refers to the constant region of an antibody excluding the first constant region immunoglobulin domain. Thus, e.g., for human immunoglobulins, “Fc” refers to the last two constant region immunoglobulin domains of IgA, IgD, and IgG, and the last three constant region immunoglobulin domains of IgE and IgM, and the flexible hinge N- terminal to these domains. For IgA and IgM, Fc may include the J chain. For IgG, Fc comprises immunoglobulin domains Cy2 and Cy3 and the hinge between Cyl and Cy2. It is understood in the art that the boundaries of the Fc region may vary, however, the human IgG heavy chain Fc region is usually defined to comprise residues C226 or P230 to its carboxylterminus, using the numbering according to the EU index as in Kabat et al. (1991, NIH Publication 91-3242, National Technical Information Service, Springfield, Va.). The term “Fc region” may refer to this region in isolation or this region in the context of an antibody or antibody fragment. “Fc region” includes naturally occurring allelic variants of the Fc region as well as modified Fc regions, e.g., that are modified to modulate effector function or other properties such as pharmacokinetics, stability or production properties of an antibody. Fc regions also include variants that do not exhibit alterations in biological function. For example, one or more amino acids can be deleted from the N-terminus or C-terminus of the Fc region of an immunoglobulin without substantial loss of biological function. Such variants can be selected according to general rules known in the art to have minimal effect on activity (see, e.g., Bowie et al., Science 247:306-1310, 1990). For example, for IgG4 antibodies, a single amino acid substitution (S228P according to Kabat numbering; designated IgG4Pro) may be introduced to abolish the heterogeneity observed in recombinant IgG4 antibodies (see, e.g., Angal et al., Mol Immunol 30:105-108, 1993).

[0108] An “EC50” as used herein refers to the half-maximal effective concentration, which is the concentration of an antibody that induces a response (signal generated in engagement33MF-363628402Attorney Docket No. 22419-20010.40 assay) halfway between the baseline and maximum after a specified exposure time. In some embodiments, the “fold over EC50” is determined by dividing the EC50 of a reference antibody by the EC50 of the test antibody.

[0109] The term “equilibrium dissociation constant” abbreviated (KD), refers to the dissociation rate constant (kd, time1) divided by the association rate constant (ka, time1M1). Equilibrium dissociation constants can be measured using any method.

[0110] The term “monovalent molecule” refers to a molecule having one antigen-binding site, e.g., a Fab or scFv.

[0111] The term “bivalent molecule” as used herein, refers to a molecule having two antigenbinding sites. In some embodiments, a bivalent molecule of the present disclosure is a bivalent antibody or a bivalent fragment thereof. In some embodiments, a bivalent molecule of the present disclosure is a bivalent antibody. In some embodiments, a bivalent molecule of the present disclosure is an IgG. In general, monoclonal antibodies have a bivalent basic structure. IgG and IgE have only one bivalent unit, while IgA and IgM consist of multiple bivalent units (2 and 5, respectively) and thus have higher valencies. This bivalency increases the avidity of antibodies for antigens.

[0112] The terms “monovalent binding” or “monovalently binds to” as used herein refer to the binding of one antigen-binding site to its antigen.

[0113] The terms “bivalent binding” or “bivalently binds to” refer to the binding of both antigen-binding sites of a bivalent molecule to its antigen. In some embodiments, both antigen-binding sites of a bivalent molecule share the same antigen specificity.

[0114] The term “valency” refers to the number of different binding sites of an antibody for an antigen. A monovalent antibody comprises one binding site for an antigen. A bivalent antibody comprises two binding sites for the same antigen.

[0115] The term “avidity” in the context of antibody binding to an antigen refers to the combined binding strength of multiple binding sites of the antibody. Thus, “bivalent avidity” refers to the combined strength of two binding sites.

[0116] The terms “identical” or percent “identity,” in the context of two or more polynucleotide or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues that are the same (e.g., 100% identity) or have a specified percentage of nucleotides or amino acid residues are the same (e.g., at least 70%, at least 75%, at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher sequence identity; or 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity)) identity over a34MF-363628402Attorney Docket No. 22419-20010.40 specified region, e.g., the length of the two sequences, when compared and aligned for maximum correspondence over a comparison window or designated region. Alignment for purposes of determining percent amino acid sequence identity can be performed in various methods, including those using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Examples of algorithms that are suitable for determining percent sequence identity and sequence similarity include the BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990). Thus, for purposes of this disclosure, BLAST 2.0 can be used with the default parameters to determine percent sequence identity.

[0117] The terms “corresponding to,” “determined with reference to,” or “numbered with reference to” when used in the context of the identification of a given amino acid residue in a polypeptide sequence, refers to the position of the residue of a specified reference sequence when the given amino acid sequence is maximally aligned and compared to the reference sequence. The polypeptide that is aligned to the reference sequence need not be the same length as the reference sequence.

[0118] A “conservative” substitution refers to a substitution of an amino acid such that charge, polarity, hydropathy (hydrophobic, neutral, or hydrophilic), and / or size of the side group chain is maintained. Illustrative sets of amino acids that may be substituted for one another include (i) positively-charged amino acids Lys and Arg; and His at pH of about 6; (ii) negatively charged amino acids Glu and Asp; (iii) aromatic amino acids Phe, Tyr and Trp; (iv) nitrogen ring amino acids His and Trp; (v) aliphatic hydrophobic amino acids Ala, Vai, Leu and He; (vi) hydrophobic sulfur-containing amino acids Met and Cys, which are not as hydrophobic as Vai, Leu, and He; (vii) small polar uncharged amino acids Ser, Thr, Asp, and Asn (viii) small hydrophobic or neutral amino acids Gly, Ala, and Pro; (ix) amide-comprising amino acids Asn and Gin; and (xi) beta- branched amino acids Thr, Vai, and lie. Reference to the charge of an amino acid in this paragraph refers to the charge at pH 6-7.

[0119] The terms “polynucleotide,” “oligonucleotide,” and “nucleic acid” are used interchangeably to refer to a polymeric form of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof, either in single-, double-, or multi- stranded form. Contemplated polynucleotides include a gene or fragment thereof. Exemplary polynucleotides include, but are not limited to, DNA, RNA, coding or non-coding regions of a gene or gene fragment, loci (locus) defined from linkage analysis, exons, introns, messenger RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), short interfering 35MF-363628402Attorney Docket No. 22419-20010.40RNA (siRNA), short-hairpin RNA (shRNA), micro-RNA (miRNA), ribozymes, cDNA, recombinant polynucleotides, branched polynucleotides, plasmids, vectors, cell-free polynucleotides including cell-free DNA (cfDNA) and cell-free RNA (cfRNA), nucleic acid probes, and primers. In a polynucleotide when referring to a T, a T means U (Uracil) in RNA and T (Thymine) in DNA. A polynucleotide can be exogenous or endogenous to a cell and / or exist in a cell-free environment. The term polynucleotide encompasses modified polynucleotides (e.g., altered backbone, sugar, or nucleobase). If present, modifications to the nucleotide structure are imparted before or after assembly of the polymer. Non-limiting examples of modifications include: 5-bromouracil, peptide nucleic acid, xeno nucleic acid, morpholines, locked nucleic acids, glycol nucleic acids, threose nucleic acids, dideoxynucleotides, cordycepin, 7-deaza-GTP, fluorophores (e.g., rhodamine or fluorescein linked to the sugar), thiol-containing nucleotides, biotin-linked nucleotides, fluorescent base analogs, CpG islands, methyl-7-guanosine, methylated nucleotides, inosine, thiouridine, pseudouridine, dihydrouridine, queuosine, and wyosine. The sequence of nucleotides may be interrupted by non-nucleotide components.

[0120] The term “vector,” as used herein, refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. The term includes the vector as a selfreplicating nucleic acid structure as well as the vector incorporated into the genome of a host cell into which it has been introduced. A “vector” as used herein refers to a recombinant construct in which a nucleic acid sequence of interest is inserted into the vector. Certain vectors can direct the expression of nucleic acids to which they are operatively linked. Such vectors are referred to herein as “expression vectors”.

[0121] A “substitution” denotes the replacement of one or more amino acids or nucleotides by different amino acids or nucleotides, respectively.

[0122] An “isolated” nucleic acid refers to a nucleic acid molecule that has been separated from a component of its natural environment. An isolated nucleic acid includes a nucleic acid molecule contained in cells that ordinarily contain the nucleic acid molecule, but the nucleic acid molecule is present extrachromosomally or at a chromosomal location that is different from its natural chromosomal location.

[0123] “Isolated nucleic acid encoding an antibody or fragment thereof’ refers to one or more nucleic acid molecules encoding antibody heavy or light chains (or fragments thereof), including such nucleic acid molecule(s) in a single vector or separate vectors, and such nucleic acid molecule(s) present at one or more locations in a host cell.36MF-363628402Attorney Docket No. 22419-20010.40

[0124] The terms “host cell,” “host cell line,” and “host cell culture” are used interchangeably and refer to cells into which exogenous nucleic acid has been introduced, including the progeny of such cells. Thus, a host cell is a recombinant host cell and includes the primary transformed cell and progeny derived therefrom without regard to the number of passages.

[0125] The term “cancer cell” or “tumor cell” as used herein refers to a neoplastic cell. The term includes cells from tumors that are benign as well as malignant. Neoplastic transformation is associated with phenotypic changes of the tumor cell relative to the cell type from which it is derived. The changes can include loss of contact inhibition, morphological changes, and unregulated cell growth.

[0126] The terms “inhibiting growth of a tumor” and “inhibiting growth of a cancer” are interchangeable and refer to slowing growth and / or reducing the cancer cell burden of a patient that has cancer. “Inhibiting growth of a cancer” thus includes killing cancer cells, as well as decreasing the rate of tumor growth, tumor size, invasion, and / or metastasis by direct or indirect effects on tumor cells.

[0127] As used herein, the terms “treatment,” “treating,” and the like, in some embodiments, refer to administering an agent, or carrying out a procedure, for the purposes of obtaining an effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or may be therapeutic in terms of affecting a partial or complete cure for a disease and / or symptoms of the disease. “Treatment,” as used herein, may include treatment of a disease or disorder (e.g., cancer) in a mammal, particularly in a human, and includes: (a) preventing the disease or a symptom of a disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it (e.g., including diseases that may be associated with or caused by a primary disease; (b) inhibiting the disease, i.e., arresting its development; and (c) relieving the disease, i.e., causing regression of the disease. Treating may refer to any indicia of success in the treatment or amelioration or prevention of a cancer, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms; or making the disease condition more tolerable to the patient; slowing in the rate of degeneration or decline; or making the final point of degeneration less debilitating. The treatment or amelioration of symptoms is based on one or more objective or subjective parameters; including the results of an examination by a physician. Accordingly, the term “treating” includes the administration of the compounds or agents of the present disclosure to prevent, delay, alleviate, arrest or inhibit development of the symptoms or conditions associated with diseases (e.g., cancer). The term "therapeutic37MF-363628402Attorney Docket No. 22419-20010.40 effect" refers to the reduction, elimination, or prevention of the disease, symptoms of the disease, or side effects of the disease in the subject.

[0128] The terms “subject,” “recipient,” “individual,” “host,” and “patient,” are used interchangeably herein and in some embodiments, refer to any mammalian subject for whom diagnosis, treatment, or therapy is desired, particularly humans. “Mammal” for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and laboratory, zoo, sports, or pet animals, such as dogs, horses, cats, cows, sheep, goats, pigs, mice, rats, rabbits, guinea pigs, monkeys etc. In some embodiments, the mammal is human. None of these terms require the supervision of medical personnel.

[0129] A “variant” of a reference antibody refers to an antibody that typically differs from the reference antibody in one or more substitutions, deletions, additions, and / or insertions in the amino acid sequence of the heavy and / or light chain.ANTI-NCAM1 ANTIBODIES

[0130] Described herein, in certain embodiments, are antibodies and antigen-binding fragments thereof that bind NCAM1 (i.e., anti-NCAMl antibodies) comprising a) a heavy chain variable region (VH) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1, (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3; and b) a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 4, (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

[0131] Further described herein, in certain embodiments, are antibodies and antigen-binding fragments thereof that bind NCAM1 (i.e., anti-NCAMl antibodies) comprising: a) a heavy chain variable region (VH) comprising 96% sequence identity to SEQ ID NO: 7 or 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises substitutions at amino acid positions 74 (Kabat) and / or 77 (Kabat)relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the substitutions are substitutions of amino acids that are not positively charged. In some embodiments, the substitutions at amino acid positions 74 (Kabat) and / or 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12, either alone or in combination, could improve the pharmacokinetic properties of the anti-NCAMl antibody or antigen-binding fragment thereof. In some embodiments, the anti-NCAMl antibody or antigen-binding fragment38MF-363628402Attorney Docket No. 22419-20010.40 thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a substitution at amino acid position 74 (Kabat) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the anti-NCAM antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises substitutions at amino acid position 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the anti-NCAM 1 antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises substitutions at amino acid position 74 (Kabat) and 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a threonine (T) at amino acid position 74 (Kabat). In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a glycine (G) at amino acid position 77 (Kabat). In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a threonine (T) at amino acid position 74 (Kabat) and a glycine (G) at amino acid position 77 (Kabat).

[0132] Further described herein, in certain embodiments, are antibodies and antigen-binding fragments thereof that bind NCAM1 (i.e., anti-NCAMl antibodies) comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 739MF-363628402Attorney Docket No. 22419-20010.40 or at least about 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises substitutions at amino acid positions 79 (sequential), and / or 82 (sequential) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the substitutions are substitutions of amino acids that are not positively charged. In some embodiments, the substitutions at amino acid positions 79 (sequential), and / or 82 (sequential) relative to the reference sequence of SEQ ID NO: 12, either alone or in combination, could improve the pharmacokinetic properties of the anti- NCAM1 antibody or antigen-binding fragment thereof. In some embodiments, the anti- NCAM1 antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a substitution at amino acid position 79 (sequential) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises substitutions at amino acid position 82 (sequential) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the anti- NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises substitutions at amino acid position 79 (sequential) and position 82 (sequential) relative to the reference sequence of SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a threonine (T) at amino acid position 79 (sequential). In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ40MF-363628402Attorney Docket No. 22419-20010.40ID NO: 12, and wherein the VL comprises a glycine (G) at amino acid position 82 (sequential). In some embodiments, the anti-NCAMl antibody or antigen-binding fragment thereof comprises: a) a VH comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a VL comprising an amino acid sequence having at least about 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein the VL comprises a threonine (T) at amino acid position 79 (sequential) and a glycine (G) at amino acid position 82 (sequential).

[0133] In some embodiments, an anti-NCAMl antibody comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1 or a variant CDR1 in which 1, 2, 3, 4, or 5 amino acids are substituted relative to the sequence; a CDR2 having an amino acid sequence according to SEQ ID NO: 2, or a variant CDR2 in which 1, 2, 3, 4, or 5 amino acids are substituted relative to the sequence; and a CDR3 having an amino acid sequence according SEQ ID NO: 3 or a variant CDR3 in which 1, 2, 3, 4, or 5 amino acids are substituted relative to the sequence. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising: a CDR1 having an amino acid sequence according to SEQ ID NO: 4, or a variant CDR1 in which 1, 2, 3, 4, or 5 amino acids are substituted relative to the sequence; a CDR2 having an amino acid sequence according to SEQ ID NO: 5, or variant CDR2 in which 1, 2, or 3 amino acids are substituted relative to the sequence; and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, or a variant CDR3 in which 1, 2, 3, 4, or 5 amino acids are substituted relative to the sequence.

[0134] In some embodiments, the anti-NCAMl antibodies comprise a) a heavy chain variable region (VH) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1, (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3; and b) a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 4, (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

[0135] In some embodiments an anti-NCAMl antibody comprises a heavy chain variable (VH) region and a light chain variable (VL) region. The VH region has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to SEQ ID NO: 7-8; and comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, or the CDR1 of SEQ ID NO: 1 in which 1, 2, 3, 4, or 5 amino acids are substituted; a CDR2 having an amino acid sequence according to SEQ ID NO: 2, or the CDR2 of SEQ ID NO: 2 in which 1, 2, 3, 4, or 5 amino acids are substituted; a41MF-363628402Attorney Docket No. 22419-20010.40CDR3 having an amino acid sequence according to SEQ ID NO: 3 or the CDR3 of SEQ ID NO: 3 in which 1, 2, 3, 4, or 5 amino acids are substituted. The VL region has an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% identical to SEQ ID NO: 9-10, and comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 4 or the CDR1 of SEQ ID NO: 4 in which 1, 2, 3, 4, or 5 amino acids are substituted; a CDR2 having an amino acid sequence according to SEQ ID NO: 5, or the CDR2 of SEQ ID NO: 5 in which 1, 2, or 3 amino acids are substituted; a CDR3 having an amino acid sequence according to SEQ ID NO: 6 or the CDR3 of SEQ ID NO: 6 in which 1, 2, 3, 4, or 5 amino acids are substituted.

[0136] In some embodiments an anti-NCAMl antibody comprises a VH and a VL, wherein the VHhas an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or 100% identical to any one of SEQ ID NOs: 7, 8, or 11; and comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, or the CDR1 of SEQ ID NO: 1 in which 1, 2, 3, 4, or 5 amino acids are substituted; a CDR2 having an amino acid sequence according to SEQ ID NO: 2, or the CDR2 of SEQ ID NO: 2 in which 1, 2, 3, 4, or 5 amino acids are substituted; a CDR3 having an amino acid sequence according to SEQ ID NO: 3 or the CDR3 of SEQ ID NO: 3 in which 1, 2, 3, 4, or 5 amino acids are substituted; and wherein the VL has an amino acid sequence that is at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or 100% identical to any one of SEQ ID NOs: 9, 10, or 12, and comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 4 or the CDR1 of SEQ ID NO: 4 in which 1, 2, 3, 4, or 5 amino acids are substituted; a CDR2 having an amino acid sequence according to SEQ ID NO: 5, or the CDR2 of SEQ ID NO: 5 in which 1, 2, 3, 4, or 5 amino acids are substituted; a CDR3 having an amino acid sequence according to SEQ ID NO: 6 or the CDR3 of SEQ ID NO: 6 in which 1, 2, 3, 4, or 5 amino acids are substituted.

[0137] In some embodiments, an anti-NCAMl antibody comprises: a VH region comprising the amino acid sequence of SEQ ID NO: 7-8 and a VL region comprising the amino acid sequence of SEQ ID NO: 9-10.

[0138] In some embodiments, an anti-NCAMl antibody comprises: a VH region comprising the amino acid sequence of SEQ ID NO: 7, 8 or 11 and a VL region comprising the amino acid sequence of SEQ ID NO: 9, 10 or 12.

[0139] In some embodiments, an anti-NCAMl antibody of the present disclosure has one, two, or three CDRs of a VH sequence having an amino acid sequence according to a42MF-363628402Attorney Docket No. 22419-20010.40 sequence set forth in Table 1. In some embodiments, the anti-NCAMl antibody has at least one mutation and no more than 10, 20, 30, 40 or 50 mutations in the VH amino acid sequences compared to a VH sequence set forth in Table 3. In some embodiments, the anti- NCAMl antibody has a VH amino acid sequence set forth in Table 3. In some embodiments, the VH amino acid sequence may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid insertions or deletions compared to a VH sequence set forth in Table 3. In some embodiments, the VH amino acid sequence may comprise a deletion or insertion, e.g., a 1, 2, 3, 4, 5, 6, or 7 amino acid deletion or insertion, relative to a CDR sequence shown in Table 1. In some embodiments, the VH comprises a CDR1 having 1 or 2 substitutions in relative to a CDR1 sequence shown in Table 1. In some embodiments, a CDR1 has 3, 4, or 5 substitutions relative to a CDR1 sequence shown in Table 1. In some embodiments, the VH comprises a CDR2 that has 1, 2, or 3 substitutions relative to the CDR2 sequence shown in Table 1. In some embodiments, the VH comprises a CDR3 that has 1, 2, 3, or 4 substitutions relative to a CDR3 sequence shown in Table 1. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 70% identity to a CDR1, CDR2, and CDR3 as shown in Table 1. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 75% identity to a CDR1, CDR2, and CDR3 as shown in Table 1. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 80% identity to a CDR1, CDR2, and CDR3 as shown in Table 1. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 85% identity to a CDR1, CDR2, and CDR3 as shown in Table 1. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 90% identity to a CDR1, CDR2, and CDR3 as shown in Table 1. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 95% identity to a CDR1, CDR2, and CDR3 as shown in Table 1. In some embodiments an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having 100% identity to a CDR1, CDR2, and CDR3 as shown in Table 1.

[0140] In some embodiments, an anti-NCAMl antibody of the present disclosure has one, two, or three CDRs of a VL sequence having an amino acid sequence according to a sequence set forth in Table 2. In some embodiments, the anti-NCAMl antibody has at least one mutation and no more than 10, 20, 30, 40 or 50 mutations in the VL amino acid sequences compared to a VL sequence set forth in Table 3. In some embodiments, the anti- 43MF-363628402Attorney Docket No. 22419-20010.40NCAM1 antibody has a VL amino acid sequence set forth in Table 3. In some embodiments, the VL amino acid sequence may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid insertions or deletions compared to a VL sequence set forth in Table 3. In some embodiments, the VL amino acid sequence may comprise a deletion or insertion, e.g., a 1, 2, 3, 4, 5, 6, or 7 amino acid deletion or insertion, relative to a CDR sequence shown in Table 2. In some embodiments, the VL region comprises a CDR1 having 1 or 2 substitutions in relative to a CDR1 sequence shown in Table 2. In some embodiments, a CDR1 has 3, 4, or 5 substitutions relative to a CDR1 sequence shown in Table 2. In some embodiments, the VL region comprises a CDR2 that has 1, 2, or 3 substitutions relative to the CDR2 sequence shown in Table 2. In some embodiments, the VL region comprises a CDR3 that has 1, 2, 3, or 4 substitutions relative to a CDR3 sequence shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 70% identity to a CDR1, CDR2, and CDR3 as shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 75% identity to a CDR1, CDR2, and CDR3 as shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 80% identity to a CDR1, CDR2, and CDR3 as shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 85% identity to a CDR1, CDR2, and CDR3 as shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 90% identity to a CDR1, CDR2, and CDR3 as shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having at least 95% identity to a CDR1, CDR2, and CDR3 as shown in Table 2. In some embodiments, an anti-NCAMl antibody of the present disclosure comprises a CDR1, CDR2, and CDR3, each having 100% identity to a CDR1, CDR2, and CDR3 as shown in Table 2.

[0141] In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 70% identical to any one of SEQ ID NO: 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 75% identical to any one of SEQ ID NO: 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 80% identical to any one of SEQ ID NO: 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 85% identical to any one of SEQ ID NO:44MF-363628402Attorney Docket No. 22419-20010.407-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 90% identical to any one of SEQ ID NO 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 95% identical to any one of SEQ ID NO 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 96% identical to any one of SEQ ID NO 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 97% identical to any one of SEQ ID NO 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 98% identical to any one of SEQ ID NO 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence at least 99% identical to any one of SEQ ID NO 7-8. In some embodiments, an anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid identical to any one of SEQ ID NO: 7-8.

[0142] In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 70% identical to any one of SEQ ID NO9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 75% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 80% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 85% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 90% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 95% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 96% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 97% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 98% identical to any one of SEQ ID NO: 9-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence at least 99% identical to any one of SEQ ID NO: 45MF-363628402Attorney Docket No. 22419-20010.409-10. In some embodiments, an anti-NCAMl antibody comprises a light chain variable region comprising an amino acid sequence identical to any one of SEQ ID NOs: 9-10.

[0143] In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region (VH) that comprises an amino acid sequence at least 60% (e.g., at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the heavy chain variable region (VH) of an anti-NCAMl antibody disclosed in Table 3, and a light chain variable region (VL) that comprises an amino acid sequence at least 60% (e.g., at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%) identical to the light chain variable region (VL) of the same anti-NCAMl antibody disclosed in Table 3.

[0144] In some embodiments, the anti-NCAMl antibody comprises a VH that comprises an amino acid sequence at least about 60% (e.g., at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) identical to the VH of an anti-NCAMl antibody disclosed in Table 3, and a VL that comprises an amino acid sequence at least about 60% (e.g., at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) identical to the VL of the same anti-NCAMl antibody disclosed in Table 3.

[0145] In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9 or 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9-10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8;46MF-363628402Attorney Docket No. 22419-20010.40 and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to any one of SEQ ID NO: 9-10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9-10. In some embodiments, the anti- NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9-10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9-10. In some embodiments, the Anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9-10. In some embodiments, the anti- NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according any one of SEQ ID NOs: 9-10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to any one of SEQ ID NOs: 7-8; and a light chain variable region comprising an amino acid sequence according to any one of SEQ ID NOs: 9-10.

[0146] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 7, 8, or 47MF-363628402Attorney Docket No. 22419-20010.4011; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to any one of SEQ ID NOs: 9, 10, or 12.

[0147] In some embodiments, the anti-NCAMl antibody comprises one, two, three, four, five, six, seven, or eight framework (FW) region sequences selected from the group consisting of: (a) a FWH1 comprising an amino acid sequence of any one of SEQ ID NOs: 19, 27, or 32, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid insertions, deletions, or substitutions; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid insertions, deletions, or substitutions; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21 or 49, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid insertions, deletions, or substitutions; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid insertions, deletions, or substitutions; (e) a FWL1 comprising an amino acid sequence of any one of SEQ ID NOs: 23 or 28, or a variant thereof comprising up to about 5 (such as about any of 1,2, 3, 4, or 5) amino acid insertions, deletions, or substitutions; (f) a FWL2 comprising an amino acid sequence of any one of SEQ ID NOs: 24, 29, or 33, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid insertions, deletions, or substitutions; (g) a FWL3 comprising an amino acid sequence of any one of SEQ ID NOs: 25 or 30, or a variant thereof comprising up to about (such as about any of 1, 2,3, 4, or 5) amino acid insertions, deletions, or substitutions; and (h) a FWL4 comprising an amino acid sequence of any one of SEQ ID NOs: 26, 31, or 34, or a variant thereof comprising up to about 5 (such as about any of 1, 2, 3, 4, or 5) amino acid insertions, deletions, or substitutions.

[0148] In some embodiments, the anti-NCAMl antibody comprises one, two, three, four, five, six, seven, or eight FW region sequences selected from the group consisting of: (a) a FWH1 comprising an amino acid sequence of any one of SEQ ID NOs: 19, 27, or 32; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of any one of SEQ ID NOs: 21 or 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of any one of SEQ ID NOs: 23 or 28; (f) a FWL2 comprising an amino acid sequence of any one of SEQ 48MF-363628402Attorney Docket No. 22419-20010.40ID NOs: 24, 29, or 33; (g) a FWL3 comprising an amino acid sequence of any one of SEQ ID NOs: 25 or 30; and (h) a FWL4 comprising an amino acid sequence of any one of SEQ ID NOs: 26, 31, or 34.

[0149] In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of any one of SEQ ID NOs: 19, 27, or 32; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of any one of SEQ ID NOs: 21 or 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of any one of SEQ ID NOs: 23 or 28; (f) a FWL2 comprising an amino acid sequence of any one of SEQ ID NOs: 24, 29, or 33; (g) a FWL3 comprising an amino acid sequence of any one of SEQ ID NOs: 25 or 30; and (h) a FWL4 comprising an amino acid sequence of any one of SEQ ID NOs: 26, 31, or 34. The amino acid sequences of the FW regions of the anti-NCAMl antibodies of the present disclosure are shown in Tables 5-6.

[0150] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 19; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 24; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and / or49MF-363628402Attorney Docket No. 22419-20010.40(h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 19; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 24; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain50MF-363628402Attorney Docket No. 22419-20010.40 variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain (HC) comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 13; and a light chain (LC) comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 14. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 13; and a LC comprising an amino acid sequence according to SEQ ID NO: 14. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0151] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence51MF-363628402Attorney Docket No. 22419-20010.40 according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 27; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 28; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 29; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 30; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 27; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 28; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 29; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 30; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments,52MF-363628402Attorney Docket No. 22419-20010.40 the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 15; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 16. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 15; and a LC comprising an amino acid sequence according to SEQ ID NO: 16. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0152] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about53MF-363628402Attorney Docket No. 22419-20010.4095%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 19; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 28; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 29; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 30; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 19; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 28; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 29; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 30; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to54MF-363628402Attorney Docket No. 22419-20010.40SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 13; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 16. In some55MF-363628402Attorney Docket No. 22419-20010.40 embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 13; and a LC comprising an amino acid sequence according to SEQ ID NO: 16. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0153] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 27; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 24; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 27; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 24; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and (h) a FWL4 comprising56MF-363628402Attorney Docket No. 22419-20010.40 an amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 99% sequence57MF-363628402Attorney Docket No. 22419-20010.40 identity with an amino acid sequence according SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 15; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 14. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 15; and a LC comprising an amino acid sequence according to SEQ ID NO: 14. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0154] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 32; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of58MF-363628402Attorney Docket No. 22419-20010.40SEQ ID NO: 24; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 32; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 24; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence59MF-363628402Attorney Docket No. 22419-20010.40 according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 17; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 14. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 17; and a LC comprising an amino acid sequence according to SEQ ID NO: 14. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0155] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino 60MF-363628402Attorney Docket No. 22419-20010.40 acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 32; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 28; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 29; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 30; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 32; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 28; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 29; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 30; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 31. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 95% sequence61MF-363628402Attorney Docket No. 22419-20010.40 identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 11; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 17; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 16. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 17; and a LC comprising an amino acid sequence according to SEQ ID NO: 16. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0156] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, 62MF-363628402Attorney Docket No. 22419-20010.40 at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 19; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 33; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 34. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 19; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 21; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 33; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 34. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid63MF-363628402Attorney Docket No. 22419-20010.40 sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 7; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 13; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more)64MF-363628402Attorney Docket No. 22419-20010.40 sequence identity with an amino acid sequence according to SEQ ID NO: 18. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 13; and a LC comprising an amino acid sequence according to SEQ ID NO: 18. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0157] In some embodiments, the anti-NCAMl antibody comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or 100%) sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a VH comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and a VL comprising a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6, and one or more FW region sequences selected from the group consisting of (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 27; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 33; (g) a FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and / or (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 34. In some embodiments, the anti-NCAMl antibody comprises (a) a FWH1 comprising an amino acid sequence of SEQ ID NO: 27; (b) a FWH2 comprising an amino acid sequence of SEQ ID NO: 20; (c) a FWH3 comprising an amino acid sequence of SEQ ID NO: 49; (d) a FWH4 comprising an amino acid sequence of SEQ ID NO: 22; (e) a FWL1 comprising an amino acid sequence of SEQ ID NO: 23; (f) a FWL2 comprising an amino acid sequence of SEQ ID NO: 33; (g) a65MF-363628402Attorney Docket No. 22419-20010.40FWL3 comprising an amino acid sequence of SEQ ID NO: 25; and (h) a FWL4 comprising an amino acid sequence of SEQ ID NO: 34. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 80% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 85% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 90% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 95% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 96% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 97% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence having at least 98% sequence identity with an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according to SEQ ID NO: 8; and 66MF-363628402Attorney Docket No. 22419-20010.40 a light chain variable region comprising an amino acid sequence having at least 99% sequence identity with an amino acid sequence according SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 8; and a light chain variable region comprising an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 15; and a LC comprising an amino acid sequence having at least about 80% (e.g., 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity with an amino acid sequence according to SEQ ID NO: 18. In some embodiments, the anti-NCAMl antibody comprises an HC comprising an amino acid sequence according to SEQ ID NO: 15; and a LC comprising an amino acid sequence according to SEQ ID NO: 18. In some embodiments, the anti-NCAMl antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0158] In some embodiments, an anti-NCAMl antibodies or antigen-binding fragments thereof described herein are humanized.

[0159] The sequences of the anti-NCAMl antibodies described herein are shown in Tables 1-7.Table 1. VH CDR sequences67MF-363628402Attorney Docket No. 22419-20010.40Table 2. VL CDR sequencesTable 3. Heavy and light variable region sequences68MF-363628402Attorney Docket No. 22419-20010.40Table 4. Heavy and light chain sequences69MF-363628402Attorney Docket No. 22419-20010.4070MF-363628402Attorney Docket No. 22419-20010.4071MF-363628402Attorney Docket No. 22419-20010.40Table 5. Framework sequence identifiersTable 6. Framework sequences72MF-363628402Attorney Docket No. 22419-20010.4073MF-363628402Attorney Docket No. 22419-20010.40Table 7. Additional CDR sequences

[0160] In some embodiments, the anti-NCAMl antibodies disclosed herein comprise one or more mutations in the VH and / or VL to remove positively charged amino acid residues. In some embodiments, the one or more mutations are R96Q and K97Q in the VH, and K50Q in the VL, wherein the positions are based on Kabat numbering. In some embodiments, the anti- NCAMl antibodies disclosed herein comprise a glutamine (Q) at amino acid position 96 in the VH, wherein the position is based on Kabat numbering. In some embodiments, the anti- NCAMl antibodies disclosed herein comprise a glutamine (Q) at amino acid position 97 in the VH, wherein the position is based on Kabat numbering. In some embodiments, the anti- NCAMl antibodies disclosed herein comprise a glutamine (Q) at amino acid position 50 in the VL, wherein the position is based on Kabat numbering. In some embodiments, the anti- NCAMl antibodies disclosed herein comprise a glutamine (Q) at amino acid position 96 in the VH, and a glutamine (Q) at amino acid position 50 in the VL, wherein the positions are based on Kabat numbering. In some embodiments the anti-NCAMl antibody comprises a VH and a VL, wherein the VH comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, and 36, and wherein the VL comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6.74MF-363628402Attorney Docket No. 22419-20010.40

[0161] In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a mutation in the DG isomerization motif at D28 and / or G29 of the VL, wherein the position is based on Kabat numbering. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a glutamic acid (E) or a serine (S) at amino acid position 28 in the VL, wherein the position is based on Kabat numbering. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a valine (V) at amino acid position 29 in the VL, wherein the position is based on Kabat numbering. In some embodiments the anti-NCAMl antibody comprises a VH and a VL, wherein the VH comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and wherein the VL comprises a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 38-40, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

[0162] In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a mutation in the NT deamidation motif at N30 and / or T31 of the VL, wherein the position is based on Kabat numbering. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a serine (S) at amino acid position 30 in the VL, wherein the position is based on Kabat numbering. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a valine (V) at amino acid position 31 in the VL, wherein the position is based on Kabat numbering. In some embodiments the anti-NCAMl antibody comprises a VH and a VL, wherein the VH comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 3, and wherein the VL comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 41 or 42, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

[0163] In some embodiments, the anti-NCAMl antibodies disclosed herein comprise one or more mutations in the methionine oxidation sites at M95 and Ml 00a of the VH, wherein the position is based on Kabat numbering. In some embodiments, the one or more mutations are M95D or M95L, and / or MIOOaL or MIOOaF in the VH, wherein the positions are based on Kabat numbering. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise an aspartic acid (D) or a leucine (L) at amino acid position 95 in the VH, wherein the position is based on Kabat numbering. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a leucine (L) or phenylalanine (F) at amino acid position 100a in75MF-363628402Attorney Docket No. 22419-20010.40 the VH, wherein the position is based on Kabat numbering. In some embodiments, the anti- NCAM1 antibodies disclosed herein comprise a leucine (L) at amino acid position 95 and a leucine (L) at amino acid position 100a in the VH, wherein the positions are based on Kabat numbering. In some embodiments the anti-NCAMl antibody comprises a VH and a VL, wherein the VH comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 43-47, and wherein the VL comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 4, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

[0164] In some embodiments, the anti-NCAMl antibodies disclosed herein comprise one or more mutations in the DG isomerization motif, NT deamidation motif, and / or methionine oxidation sites. In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a leucine (L) at amino acid position 95 and a leucine (L) at amino acid position 100a in the VH, and a valine (V) at amino acid position 29 and a serine (S) at amino acid position 30 in the VL, wherein the position is based on Kabat numbering. In some embodiments the anti- NCAMl antibody comprises a VH and a VL, wherein the VH comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 1, a CDR2 having an amino acid sequence according to SEQ ID NO: 2, and a CDR3 having an amino acid sequence according to SEQ ID NO: 47, and wherein the VL comprises a CDR1 having an amino acid sequence according to SEQ ID NO: 48, a CDR2 having an amino acid sequence according to SEQ ID NO: 5, and a CDR3 having an amino acid sequence according to SEQ ID NO: 6.Fc variants

[0165] In some embodiments, the anti-NCAMl antibodies disclosed herein comprise a modified Fc region (as further explained herein). Fc regions typically comprises one or more Fc chains. An IgG Fc chain typically contains two constant heavy domains (CH2 and CH3) and a hinge region connected to the CH2 domain. Fc regions may typically comprise two Fc chains which dimerize with one another; however, an Fc region may have a single chain or more than two Fc chains, e.g., as may be present in some antibody formats.

[0166] In some embodiments, the anti-NCAMl antibodies comprise an IgGl Fc region (e.g., human IgGl Fc region), that is, except for having particular residue(s) at certain positions as noted herein, the Fc region has an amino acid sequence that is substantially similar to that of the Fc region within a wild type IgGl Fc. In some embodiments, the wild type IgGl Fc is a76MF-363628402Attorney Docket No. 22419-20010.40 human IgGl. In some embodiments, the anti-NCAMl antibodies comprise an Fc region, each Fc chain of which has an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to that of an Fc chain within a wild-type IgGl Fc. In some embodiments, the anti-NCAMl antibodies comprise an Fc region, each Fc chain of which has an amino acid sequence that is at least about 85% (e.g., at least about 87.5%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical to that of an Fc chain within a wild-type IgGl Fc.

[0167] In some embodiments, the anti-NCAMl antibodies comprise an IgG2 Fc region (e.g., human IgG2 Fc region), that is, except for having particular residue(s) at certain positions as noted herein, the Fc region has an amino acid sequence that is substantially similar to that of the Fc region within a wild type IgG2 Fc. In some embodiments, the wild type IgG2 Fc is a human IgG2 Fc. In some embodiments, the anti-NCAMl antibodies comprise an Fc region, each Fc chain of which has an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to that of an Fc chain within a wild-type IgG2 Fc. In some embodiments, the anti-NCAMl antibodies comprise an Fc region, each Fc chain of which has an amino acid sequence that is at least about 85% (e.g., at least about 87.5%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical to that of an Fc chain within a wild-type IgG2 Fc.

[0168] In some embodiments, the anti-NCAMl antibodies comprise an IgG4 Fc region (e.g., human IgG4 Fc region), that is, except for having particular residue(s) at certain positions as noted herein, the Fc region has an amino acid sequence that is substantially similar to that of the Fc region within a wild type IgG4 Fc. In some embodiments, the wild type IgG4 Fc is a human IgG4 Fc. In some embodiments, the anti-NCAMl antibodies comprise an Fc region, each Fc chain of which has an amino acid sequence that is at least 85%, at least 87.5%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to that of an Fc chain within a wild-type IgG4 Fc. In some embodiments, the anti-NCAMl antibodies comprise an Fc region, each Fc chain of which has an amino acid sequence that is at least about 85% (e.g., at least about 87.5%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least77MF-363628402Attorney Docket No. 22419-20010.40 about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%) identical to that of an Fc chain within a wild-type IgG4 Fc.

[0169] In certain embodiments, Fc regions are modified (e.g., substituted) at one more amino acid residues. In certain embodiments, such modifications alter the half-life of a molecule (e.g., binding protein) which comprises the Fc region by altering (e.g., enhancing) binding to an Fc receptor such as the neonatal Fc receptor (FcRn)

[0170] Unless otherwise specified herein, numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also called the EU index, as described in Kabat et al, Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991.

[0171] In some embodiments, one or more modifications in the modified Fc region is selected from the group consisting of: S298A, E333A, K334A, K326A, F243L, R292P, Y300L, V305I, P396L, F243L, R292P, Y300L, L235V, P396L, F243L, S239D, I332E, A330L, S267E, L328F, D265S, S239E, K326A, A327H, G237F, K326E, G236A, D270L, H268D, S324T, L234F, N325L, V266L, and S267D. In some embodiments, one or more modifications in the modified Fc region is selected from the group consisting of S228P, M252Y, S254T, T256E, T256D, T250Q, H285D, T307A, T307Q, T307R, T307W, L309D, Q411H, Q311V, A378V, E380A, M428L, N434A, N434S, N297A, D265A, L234A, L235A, and N434W.

[0172] In some embodiments, the modified Fc region comprises a specific combination of amino acid substitutions selected from the group consisting of: L234A / L235A;V234A / G237A; L235A / G237A / E318A; S228P / L236E; H268Q / V309L / A330S / A331S; C220S / C226S / C229S / P238S; C226S / C229S / E3233P / L235V / L235A; L234F / L235E / P331S; C226S / P230S; L234A / G237A; L234A / L235A / G237A; Q311R / M428L; L234A / L235A (LALA); L234A / L235A / P329G (LALAPG); L234A / L235A / P329S (LALAPS); and L234A / L235A / P329A (LALAPA). In some embodiments, the modified Fc region comprises a specific combination of amino acid substitutions consisting of L234A / L235A / P329A (LALAPA). In some embodiments, the modified Fc region comprises a specific combination of amino acid substitutions consisting of L234A / L235A / P329G (LALAPG). In some embodiments, the modified Fc region comprises a specific combination of amino acid substitutions consisting of L234A / L235A / P329S (LALAPS).

[0173] In some embodiments, the modified Fc region comprises a specific combination of amino acid substitutions selected from the group consisting of M428L / N434S (LS); M252Y / S254T / T256E (YTE); T250Q / M428L; T307A / E380A / N434A; T256D / T307Q (DQ);78MF-363628402Attorney Docket No. 22419-20010.40T256D / T307W (DW); M252Y / T256D (YD); T307Q / Q311V / A378V (QVV); T256D / H285D / T307R / Q311V / A378V (DDRVV); L309D / Q311H / N434S (DHS); S228P / L235E (SPLE); L234A / L235A (LALA); M428L / N434A (LA); L234A / G237A (LAGA); L234A / L235A / G237A (LALAGA); L234A / L235A / P329G (LALAPG); L234A / L235A / P329A (LALAPA); N297A / YTE; D265A / YTE; LALA / YTE; LAGA / YTE; LALAGA / YTE; LALAPG / YTE; N297A / LS; D265A / LS; LALA / LS; LAGA / LS;LALAGA / LS; LALAPG / LS; N297A / DHS; D265A / DHS; LALA / DHS; LAGA / DHS; LALAGA / DHS; LALAPG / DHS; SP / YTE; SPLE / YTE; SP / LS; SPLE / LS; SP / DHS; SPLE / DHS; N297A / LA; D265A / LA; LALA / LA; LAGA / LA; LALAGA / LA; LALAPG / LA; N297A / N434A; D265A / N434A; LALA / N434A; LAGA / N434A; LALAGA / N434A;LALAPG / N434A; N297A / N434W; D265A / N434W; LALA / N434W; LAGA / N434W; LALAGA / N434W; LALAPG / N434W; N297A / DQ; D265A / DQ; LALA / DQ; LAGA / DQ; LALAGA / DQ; LALAPG / DQ; N297A / DW; D265A / DW; LALA / DW; LAGA / DW; LALAGA / DW; LALAPG / DW; N297A / YD; D265A / YD; LALA / YD; LAGA / YD;LALAGA / YD; LALAPG / YD; N297A / QVV; D265A / QVV; LALA / QVV; LAGA / QVV, LALAGA / QVV; LALAPG / QVV; N297A / DDRVV; D265A / DDRVV; LALA / DDRVV; LAGA / DDRVV; LALAGA / DDRVV; LALAPG / DDRVV; SP / Q311R / M428L;SPLE / Q311R / M428L; N297A / Q311R / M428L; D265A / Q311R / M428L;LALA / Q311R / M428L; LAGA / Q311R / M428L; LALAGA / Q311R / M428L; and LALAPG / Q311R / M428L.

[0174] In addition to the variants discussed above, there are several useful Fc amino acid modifications that can be made for a variety of reasons, including, but not limited to, altering binding to one or more FcyR receptors, altered binding to FcRn receptors and the like as discussed below. Accordingly, the antibodies provided herein (heterodimeric, as well as homodimeric) can include such amino acid modifications with or without the heterodimerization variants outlined herein (e.g., the pl variants and steric variants). Each set of variants can be independently and optionally included or excluded from any heterodimeric protein.FcyR Variants

[0175] Accordingly, there are several useful Fc substitutions that can be made to alter binding to one or more of the FcyR receptors. In certain embodiments, the subject antibody includes modifications that alter the binding to one or more FcyR receptors (i.e., “FcyR variants”). Substitutions that result in increased binding as well as decreased binding can be 79MF-363628402Attorney Docket No. 22419-20010.40 useful. For example, it is known that increased binding to FcyRIIIa generally results in increased ADCC (antibody dependent cell-mediated cytotoxicity; the cell-mediated reaction wherein nonspecific cytotoxic cells that express FcyRs recognize bound antibody on a target cell and subsequently cause lysis of the target cell). Similarly, decreased binding to FcyRIIb (an inhibitory receptor) can be beneficial as well in some circumstances. Amino acid substitutions that find use in the antibodies described herein include those listed in US Patent Nos. 8,188,321 (particularly Figure 41) and 8,084,582, and US Publ. App. Nos. 20060235208 and 20070148170. Particular variants that find use include, but are not limited to, 236A, 239D, 239E, 332E, 332D, 239D / 332E, 267D, 267E, 328F, 267E / 328F, 236A / 332E, 239D / 332E / 330Y, 239D / 332E / 330L, 243A, 243L, 264 A, 264V and 299T.

[0176] In addition, there are additional Fc substitutions that find use in increased binding to the FcRn receptor and increased serum half-life, as specifically disclosed in USSN 12 / 341,769 including, but not limited to, 434S, 434A, 428L, 308F, 2591, 428L / 434S, 259F308F, 436F428L, 4361, 436V / 434S, 436V / 428L and 259P308F / 428L. Such modification may be included in one or both Fc domains of the subject antibody. In certain embodiments, the anti-NCAMl antibodies comprising Fc substitutions that have increased binding to the FcRn receptor and increased serum half-life have an increased half-life that is about 10,000-fold, 1,000-fold, 500-fold, 100-fold, 50-fold, 20-fold, 10-fold, 9-fold, 8-fold, 7- fold, 6-fold, 5-fold, 4.5-fold, 4-fold, 3.5-fold, 3-fold, 2.5-fold, 2-fold, 1.95-fold, 1.9-fold, 1.85-fold, 1.8-fold, 1.75-fold, 1.7-fold, 1.65-fold, 1.6-fold, 1.55-fold, 1.50-fold, 1.45-fold, 1.4-fold, 1.35-fold, 1.3-fold, 1.25-fold, 1.2-fold, 1.15-fold, 1.1-fold, or 1.05-fold longer compared to a binding protein comprising a wild-type Fc region.

[0177] In some embodiments, an anti-NCAMl antibody disclosed herein, including antibody fragments, of the present disclosure comprises an Fc region that has effector function, e.g., exhibits antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and / or complement-dependent cytotoxicity (CDC). In some embodiments, the Fc region may be an Fc region engineered to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or ADCC. Accordingly, an Fc region can comprise additional mutations to increase or decrease effector functions, i.e., the ability to induce certain biological functions upon binding to an Fc receptor expressed on an immune cell. Immune cells include, but are not limited to, monocytes, macrophages, neutrophils, dendritic cells, eosinophils, mast cells, platelets, B cells, large granular lymphocytes, Langerhans’ cells, natural killer (NK) cells, and cytotoxic T cells.80MF-363628402Attorney Docket No. 22419-20010.40

[0178] In some embodiments, an Fc region described herein can include additional modifications that modulate effector function. Examples of Fc region amino acid mutations that modulate an effector function include, but are not limited to, one or more substitutions at positions 228, 233, 234, 235, 236, 237, 238, 239, 243, 265, 269, 270, 297, 298, 318, 326, 327, 329, 330, 331, 332, 333, and 334 (EU numbering scheme) of an Fc region.

[0179] Illustrative substitutions that decrease effector functions include the following: position 329 may have a mutation in which proline is substituted with a glycine or arginine or an amino acid residue large enough to destroy the Fc / Fcy receptor interface that is formed between proline 329 of the Fc and tryptophan residues Trp 87 and Trp 110 of FcyRIII. Additional illustrative substitutions that decrease effector functions include S228P, E233P, E235E, N297A, N297D, and P331S. Multiple substitutions may also be present, e.g., E234A and E235A of a human IgGl Fc region; E234A, E235A, P329A of a human IgGl Fc region; E234A, E235A, and P329G of a human IgGl Fc region; S228P and E235E of a human IgG4 Fc region; E234A and G237A of a human IgGl Fc region; E234A, E235A, and G237A of a human IgGl Fc region; V234A and G237A of a human IgG2 Fc region; E235A, G237A, and E318A of a human IgG4 Fc region; and S228P and E236E of a human IgG4 Fc region, to decrease effectors functions. Examples of substitutions that increase effector functions include, e.g., E333A, K326W / E333S, S239D / I332E / G236A, S239D / A330E / I332E, G236A / S239D / A330E / I332E, F243E, G236A, and S298A / E333A / K334A. In some embodiments, the Fc mutations include P329G, E234A, E235A, or a combination thereof.

[0180] In some embodiments, an Fc region may have one or more amino acid substitutions that modulate ADCC, e.g., substitutions at positions 298, 333, and / or 334 of the Fc region, according to the EU numbering scheme. Specifically, S298A, E333A, and K334A can be introduced to an Fc region to increase the affinity of the Fc region to FcyRIIIa and decrease the affinity of the Fc region to FcyRIIa and FcyRIIb.

[0181] An Fc region can also comprise additional mutations to increase serum half-life. Through enhanced binding to the neonatal Fc receptor (FcRn), such mutations in an Fc region can improve the pharmacokinetics of the antibody. Examples of substitutions in an Fc region that increase the serum half-life of an antibody include, e.g., M252Y / S254T / T256E, T250Q / M428E, N434A, N434H, T307A / E380A / N434A, M428E / N434S, M252Y / M428E, D259PV308F, N434S, V308W, V308Y, and V308F.

[0182] In some embodiments, an anti-NCAMl antibody described herein comprise an Fc region having altered glycosylation that increases the ability of the antibody to recruit NK cells and / or increase ADCC. In some embodiments, the Fc region comprises glycan81MF-363628402Attorney Docket No. 22419-20010.40 containing no fucose (i.e., the Fc region is afucosylated). Afucosylated antibodies can be produced using cell lines that express a heterologous enzyme that depletes the fucose pool inside the cell (e.g., GlymaxX® by ProBioGen AG, Berlin, Germany).

[0183] Furthermore, in some embodiments, an antibody of the disclosure may be chemically modified (e.g., one or more chemical moieties can be attached to the antibody) or be modified, e.g., produced in cell lines and / or in cell culture conditions to alter its glycosylation (e.g., hypofucosylation, afucosylation, or increased sialylation), to alter one or more functional properties of the antibody. For example, the antibody can be linked to one of a variety of polymers, for example, polyethylene glycol. In some embodiments, an antibody may comprise mutations to facilitate linkage to a chemical moiety and / or to alter residues that are subject to post-translational modifications, e.g., glycosylation.ACTIVITYNCAMl-binding activity

[0184] The activity of the anti-NCAMl antibodies as described herein can be assessed for binding in binding assays. Nonlimiting examples of suitable assays include bio-layer interferometry (BLI), surface plasmon resonance analysis using a biosensor system such as a Biacore® system, plate-based immunoassays (e.g., ELISA) or a flow cytometry assay, which are further described in the EXAMPLES section.

[0185] In some embodiments, a variant anti-NCAMl antibody in accordance with the present disclosure may block binding of the reference antibody in a competition assay by about 50% or more, about 55% or more, about 60% or more, about 65% or more, about 70% or more, about 80% or more, or about 90% or more.

[0186] In some embodiments, binding assays to assess variant activity are performed on NCAMl-expressing tissues or NCAM1 -expressing cells ex vivo. Binding can be assessed by any number of means including flow cytometry and immunohistochemistry or immunofluorescence-based assays.

[0187] In some embodiments the antibody is added to a cancer cell line and the binding is analyzed by flow cytometry. In some embodiments, the binding of the antibodies to bind to NCAMl-expressing cells are assessed by immunofluorescence methods, as described in the EXAMPLES. The anti-NCAMl antibodies preferentially bind to NCAM1 expressing cells.

[0188] In some embodiments, the binding activity of the antibodies is assessed by determining EC50 values, and in some embodiments additionally determining delta activity, i.e., the difference in specific activity between lower and upper plateaus of the activation82MF-363628402Attorney Docket No. 22419-20010.40 curve expressed as percent of activity of a selected antibody having known in vitro activity. In typical embodiments, EC50 values are compared to a reference antibody.

[0189] In some embodiments antibodies of the present disclosure have a KD of less than about 50 nM, typically less than about 25 nM, or less than 10 nM, e.g., less than about 5 nM or than about 1 nM and often less than about 10 nM as determined by surface plasmon resonance analysis using a biosensor system such as a Biacore® system performed at 37°C. In some embodiments, an antibody of the present disclosure has a KD of less than 5 x 10’5M, less than 10’5M, less than 5 x 10’6M, less than 10’6M, less than 5 x 10’7M, less than 10’7M, less than 5 x 10’8M, less than 10’8M, less than 5 x 10’9M, less than 10’9M, less than 5 xlO10M, less than 1010M, less than 5 x 1011M, less than 1011M, less than 5 x 1012M, less than 1012M, less than 5 x 1013M, less than 1013M, less than 5 x 1014M, less than 1014M, less than 5 x 1015M, or less than 1015M or lower as measured as a bivalent antibody. In some embodiments, an antibody of the present disclosure has a KD of less than 10’5to less than 10’15as measured as a bivalent antibody. In some embodiments, an antibody of the present disclosure has a KD of 10’6to IO10as measured as a bivalent antibody. In the context of the present disclosure, an “improved” KD refers to a lower KD. In some embodiments, an antibody of the present disclosure has a KD of less than 5 x 10’5M, less than 10’5M, less than 5 x 10’6M, less than 10’6M, less than 5 x 10’7M, less than 10’7M, less than 5 x 10’8M, less than 10’8M, less than 5 x 10’9M, less than 10’9M, less than 5 xlOloM, less than 1010M, less than 5 x 1011M, less than 1011M, less than 5 x 1012M, less than 1012M, less than 5 x 1013M, less than 1013M, less than 5 x 1014M, less than 1014M, less than 5 x 1015M, or less than 1015M or lower as measured as a monovalent antibody, such as a monovalent Fab. In some embodiments, an antibody of the present disclosure has a KD of less than 10’5to less than 1015as measured as a monovalent antibody, such as a monovalent Fab. In some embodiments, an antibody of the present disclosure has a KD of 10’6to IO10as measured as a monovalent antibody, such as a monovalent Fab. In some embodiments, an anti-NCAMl antibody of the present disclosure has KD less than 100 pM, e.g., or less than 75 pM, e.g., in the range of 1 to 100 pM, when measured by surface plasmon resonance analysis using a biosensor system such as a Biacore® system performed at 37°C. In some embodiments, an anti-NCAMl antibody of the present disclosure has KD of greater than 100 pM, e.g., in the range of 100-1000 pM or 500-1000 pM when measured by bio-layer interferometry (BFI) or surface plasmon resonance (SPR) analysis using a biosensor system such as a Biacore® system performed at 37 °C.83MF-363628402Attorney Docket No. 22419-20010.40

[0190] In some embodiments, the binding affinity (KD) and / or binding activity of the anti- NCAM1 antibodies can be evaluated under stress conditions (e.g., evaluating retention of binding response under an acidic stress condition as compared to an unstressed control).Fc effector function

[0191] In some embodiments, an anti-NCAMl antibody of the present disclosure comprises an Fc region that has effector function. Examples of effector functions include, but are not limited to, Clq binding and complement-dependent cytotoxicity (CDC), Fc receptor binding (e.g., FcyR binding), ADCC, antibody-dependent cell-mediated phagocytosis (ADCP), downregulation of cell surface receptors (e.g., B cell receptor), and B-cell activation. Effector functions may vary with the antibody class. For example, native human IgGl and IgG3 antibodies can elicit ADCC and CDC activities upon binding to an appropriate Fc receptor present on an immune system cell; and native human IgGl, IgG2, IgG3, and IgG4 can elicit ADCP functions upon binding to the appropriate Fc receptor present on an immune cell.

[0192] In some embodiments, the Fc region of an anti-NCAMl antibody disclosed herein may be an Fc region engineered to alter one or more functional properties of the antibody, such as serum half-life, complement fixation, Fc receptor binding, and / or ADCC. Accordingly, an Fc region can comprise additional mutations to increase or decrease effector functions, i.e., the ability to induce certain biological functions upon binding to an Fc receptor expressed on an immune cell. Immune cells include, but are not limited to, monocytes, macrophages, neutrophils, dendritic cells, eosinophils, mast cells, platelets, B cells, large granular lymphocytes, Eangerhans’ cells, natural killer (NK) cells, and cytotoxic T cells.

[0193] The anti-NCAMl antibodies of the present disclosure may be evaluated in various assays for their ability to mediate FcR-dependent activity. In one assay, the binding activity of an anti-NCAMl antibody is evaluated in an Fc receptor engagement assay. For purposes of testing variants, “engagement” of an Fc receptor occurs when a variant antibody binds to both a target NCAM1 cell via its Fv region and an FcyR present on an immune cell via the antibody Fc region in such as manner to transduce a signal. If the Fc region is kept constant among variants that differ in their Fv regions, then the assay allows an evaluation of NCAM1 binding activity across such variants in the context of potential signal transduction through a particular Fc region binding a particular Fc receptor. In some embodiments, binding of the antibody Fc region can result in clustering and / or internalization of the FcR, resulting in a luminescence signal in cells harboring a NFAT-RE-Eucif erase reporter construct.84MF-363628402Attorney Docket No. 22419-20010.40ANTIBODY FORMATS

[0194] In some embodiments, an anti-NCAMl antibody in accordance with the present disclosure is in a monovalent format. In some embodiments, the NCAM1 -targeting antibody is in a fragment format, e.g., a Fv, Fab, Fab’, scFv, diabody, or F(ab’)2 fragment. In some embodiments, the NCAM1 -targeting antibody is a nanobody (nb), minibody, small immunoprotein (SIP), or variable new antigen receptor (VNAR). In some embodiments, of the disclosure, an anti-NCAMl antibody in accordance with the disclosure may be an antibody fragment, e.g., a Fv, Fab, Fab’, scFv, VHH, diabody, or F(ab’)2 fragment. In another embodiment, the antibody is a substantially full-length antibody, e.g., an IgG antibody or other antibody class or isotype as defined herein. For a review of certain antibody fragments, see Hudson et al. Nat. Med. 9: 129-134 (2003). Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody as well as production by recombinant host cells.

[0195] In some embodiments, an anti-NCAMl antibody according to the present disclosure that is administered to a patient is an IgG of the IgGl subclass. In some embodiments, such an antibody is an IgG of the IgG2, IgG3, or IgG4 subclass. In some embodiments, such an antibody is an IgM. In some embodiments, such an antibody has a lambda light chain constant region. In some embodiments, such an antibody has a kappa light chain constant region.

[0196] In some embodiments, an anti-NCAMl antibody of the present disclosure is employed in a bispecific or multi- specific format, e.g., a tri-specific format. For example, in some embodiments, the antibody may be incorporated into a bispecific or multi- specific antibody that comprises a further binding domain that binds to the same or a different antigen.

[0197] There are a variety of possible formats that can be used in bispecific or multi-specific antibodies. The formats can vary elements such as the number of binding arms, the format of each binding arm (e.g., Fab, scFv, scFab, or VH-only), the number of antigen binding domains present on the binding arms, the connectivity and geometry of each arm with respect to each other, the presence or absence of an Fc domain, the Ig class (e.g., IgG or IgM), the Fc subclass (e.g., hlgGl, hIgG2, or hIgG4), and any mutations to the Fc (e.g., mutations to reduce or increase effector function or extend serum half-life).. Also see Speiss, et al., Alternative Molecular Formats and Therapeutic Applications for Bispecific Antibodies, Mol85MF-363628402Attorney Docket No. 22419-20010.40Immunol, 67, 95-106 (2015), particularly FIG. 1, for examples of bispecific and multispecific formats.

[0198] In some embodiments of any of the above bispecific or multispecific antibody constructs, the NCAM1 -targeting binding domain comprises all six CDRs (CDR1, CDR2, CDR3, CDR1, CDR2, and CDR3) sequences from the individual antibodies disclosed in Tables 1 and 2.

[0199] In some embodiments of any of the above bispecific or multispecific antibody constructs, the NCAM1 -targeting binding domain comprises the VH and VL sequences from the individual antibodies disclosed in Table 3.

[0200] In some embodiments of any of the above bispecific or multispecific antibody constructs, the NCAM1 -targeting binding domain comprises all six CDRs (CDR1, CDR2, CDR3, CDR1, CDR2, and CDR3) sequences from any one of antibodies Antibody 1 or Antibody 2.

[0201] In some embodiments of any of the above bispecific or multispecific antibody constructs, the NCAM1 -targeting binding domain comprises the VH and VL sequences of Antibody 1 or Antibody 2.

[0202] In some embodiments, an anti-NCAMl antibody disclosed herein is constructed as a multivalent antibody. In some embodiments an anti-NCAMl antibody is constructed as a tetravalent molecule, comprising four glycan-binding arms per molecule. Such constructs exhibit increased ADCC activity, as well as increased binding to NCAM1 cells a measured by flow cytometry.

[0203] In some embodiments, an anti-NCAMl antibody of the present disclosure is employed in a bispecific or multi- specific format, e.g., a tri-specific format. For example, in some embodiments, the antibody may be incorporated into a bispecific or multi- specific antibody that comprises a further binding domain that binds to the same or a different antigen.

[0204] There are a variety of possible formats that can be used in bispecific or multi-specific antibodies. The formats can vary elements such as the number of binding arms, the format of each binding arm (e.g., Fab, scFv, scFab, or VH-only), the number of antigen binding domains present on the binding arms, the connectivity and geometry of each arm with respect to each other, the presence or absence of an Fc domain, the Ig class (e.g., IgG or IgM), the Fc subclass (e.g., hlgGl, hIgG2, or hIgG4), and any mutations to the Fc (e.g., mutations to reduce or increase effector function or extend serum half-life).86MF-363628402Attorney Docket No. 22419-20010.40GENERATION OF ANTIBODIES

[0205] The anti-NCAMl antibodies can be produced using vectors and recombinant methodology well known in the art. Reagents, cloning vectors, and kits for genetic manipulation are available from commercial vendors. Accordingly, in some embodiments of the disclosure, provided herein are isolated nucleic acids encoding a VH and / or VL region, or fragment thereof, of any of the NCAM1 -targeting antibodies as described herein; vectors comprising such nucleic acids and host cells into which the nucleic acids are introduced that are used to replicate the antibody-encoding nucleic acids and / or to express the antibodies. Such nucleic acids may encode an amino acid sequence containing the VL and / or an amino acid sequence containing the VH of the NCAM1 -targeting antibody (e.g., the light and / or heavy chains of the antibody). In some embodiments, the host cell contains (1) a vector containing a polynucleotide that encodes the VL amino acid sequence and a polynucleotide that encodes the VH amino acid sequence, or (2) a first vector containing a polynucleotide that encodes the VL amino acid sequence and a second vector containing a polynucleotide that encodes the VH amino acid sequence.

[0206] In some embodiments, the disclosure provides a method of making an anti-NCAMl antibody as described herein. In some embodiments, the method includes culturing a host cell as described in the preceding paragraph under conditions suitable for expression of the antibody. In some embodiments, the antibody is subsequently recovered from the host cell (or host cell culture medium).

[0207] Suitable vectors containing polynucleotides encoding antibodies of the present disclosure, or fragments thereof, include cloning vectors and expression vectors. While the cloning vector selected may vary according to the host cell intended to be used, useful cloning vectors generally can self-replicate, may possess a single target for a particular restriction endonuclease, and / or may carry genes for a marker that can be used in selecting clones containing the vector. Examples include plasmids and bacterial viruses, e.g., pUC18, pUC19, Bluescript (e.g., pBS SK+) and its derivatives, mpl8, mpl9, pBR322, pMB9, ColEl plasmids, pCRl, RP4, phage DNAs, and shuttle vectors. These and many other cloning vectors are available from commercial vendors.

[0208] Expression vectors generally are replicable polynucleotide constructs that contain a nucleic acid of the present disclosure. The expression vector can be replicable in the host cells either as episomes or as an integral part of the chromosomal DNA. Suitable expression vectors include but are not limited to plasmids and viral vectors, including adenoviruses, adeno-associated viruses, retroviruses, and any other vector.87MF-363628402Attorney Docket No. 22419-20010.40

[0209] Suitable host cells for expressing an anti-NCAMl antibody as described herein include both prokaryotic and eukaryotic cells. For example, an anti-NCAMl antibody may be produced in bacteria when glycosylation and Fc effector function are not needed. After expression, the antibody may be isolated from the bacterial cell paste in a soluble fraction and can be further purified. Alternatively, the host cell may be a eukaryotic host cell, including eukaryotic microorganisms, such as filamentous fungi or yeast, including fungi and yeast strains whose glycosylation pathways have been “humanized,” resulting in the production of an antibody with a partially or fully human glycosylation pattern, vertebrate, invertebrate, and plant cells. Examples of invertebrate cells include insect cells. Numerous baculoviral strains have been identified which may be used in conjunction with insect cells. Plant cell cultures can also be utilized as host cells.

[0210] In some embodiments, vertebrate host cells are used for producing an anti-NCAMl antibody of the present disclosure. For example, mammalian cell lines such as a monkey kidney CV1 line transformed by SV40 (COS-7); human embryonic kidney line (293 or 293 cells as described, e.g., in Graham et al., J. Gen Virol. 36:59,1977; baby hamster kidney cells (BHK); mouse sertoli cells (TM4 cells as described, e.g., in Mather, Biol. Reprod. 23:243- 251, 1980 monkey kidney cells (CV1); African green monkey kidney cells (VERO-76); human cervical carcinoma cells (HELA); canine kidney cells (MDCK; buffalo rat liver cells (BRL 3A); human lung cells (W138); human liver cells (Hep G2); mouse mammary NCAM1 (MMT 060562); TRI cells, as described, e.g., in Mather et al., Annals N.Y. Acad. Sci. 383:44-68, 1982; MRC 5 cells; and FS4 cells may be used to express an NCAM1 -targeting antibodies. Other useful mammalian host cell lines include Chinese hamster ovary (CHO) cells, including DHFR- CHO cells (Urlaub et al., Proc. Natl. Acad. Sci. USA 77:4216, 1980); and myeloma cell lines such as Y0, NS0 and Sp2 / 0. Host cells of the present disclosure also include, without limitation, isolated cells, in vitro cultured cells, and ex vivo cultured cells. For a review of certain mammalian host cell lines suitable for antibody production, see, e.g., Yazaki and Wu, Methods in Molecular Biology, Vol. 248 (B.K.C. Lo, ed., Humana Press, Totowa, NJ), pp. 255-268, 2003.

[0211] In some embodiments, an anti-NCAMl antibody of the present disclosure is produced by a CHO cell line, e.g., the CHO-K1 cell line. One or more expression plasmids can be introduced that encode heavy and light chain sequences. For example, an expression plasmid encoding a heavy chain disclosed herein, and an expression plasmid encoding a light chain disclosed herein are transfected into host cells. The expression plasmids can be introduced as linearized plasmids at a ratio of 1:1 in the CHO-K1 host cell line using reagents such as88MF-363628402Attorney Docket No. 22419-20010.40Freestyle Max reagent. Fluorescence-activated cell sorting (FACS) coupled with single cell imaging can be used as a cloning method to obtain a production cell line.

[0212] A host cell transfected with an expression vector encoding an anti-NCAMl antibody of the present disclosure, or fragment thereof, can be cultured under appropriate conditions to allow expression of the polypeptide to occur. The polypeptides may be secreted and isolated from a mixture of cells and medium containing the polypeptides. Alternatively, the polypeptide may be retained in the cytoplasm or in a membrane fraction and the cells harvested, lysed, and the polypeptide isolated using a desired method.

[0213] In some embodiments, an anti-NCAMl antibody of the present disclosure can be produced by in vitro synthesis (see, e.g., Sutro Biopharma biochemical protein synthesis platform).

[0214] In some embodiments, provided herein is a method of generating variants of an anti- NCAMl antibody as disclosed herein. Thus, for example, a construct encoding a variant of a VH CDR3 as described herein can be modified and the VH region encoded by the modified construct can be tested for binding activity to NCI-H69 (ATCC catalog no. HTB-119) cells and / or in vivo NCAM1 -targeting activity in the context of a VH region as described herein, that is paired with a VL region or variant region as described herein. Similarly, a construct encoding a variant of a VL CDR3 as described herein can be modified and the VL region encoded by the modified construct can be tested for binding to NCLH69 (ATCC catalog no. HTB-119) cells, or other NCAM1 cells, and / or in vivo NCAM1 -targeting activity efficacy. Such an analysis can also be performed with other CDRs or framework regions and an antibody having the desired activity can then be selected.NCAM1-TARGETING IMMUNOCONJUGATES

[0215] Described herein, in certain embodiments, are immunoconjugates comprising anti- NCAMl antibodies described herein.

[0216] In certain embodiments, the present disclosure provides an immunoconjugate, comprising: a) an antibody or antigen-binding fragment thereof that binds to NCAM1, wherein the antibody comprises: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid89MF-363628402Attorney Docket No. 22419-20010.40 sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); and b) a topoisomerase I inhibitor conjugated to the antibody or the antigen-binding fragment thereof using a linker.

[0217] In some embodiments, the topoisomerase I inhibitor is represented by Formula (II- A) having the structure:wherein: R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and -[(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring; R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-C6haloalkyl, or — [(CY2)PO(CY2)q]tCY3; R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkylj-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci-Cehaloalkynylj-X2; X1is — O — , — S(O)W— , — NH — , — O — (C=O) — , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ; X2is — OR9, — SR9, or — NHR9; R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually H or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6; R7is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker; R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted90MF-363628402Attorney Docket No. 22419-20010.40 or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)pO(CY2)q]tCY2 — X3; R9is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker, with the proviso that exactly one of R7and R9is the point of attachment to the linker; and each X3is individually — H, —OH, — SH, or — NH2.

[0218] In some embodiments, the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein: R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and -[(CY2)PO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring; R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci-C6haloalkynyl)-X2; X1is — O — , — S(O)W— , — NH — , — O — (C=O) — , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ; X2is — OR9, — SR9, or — NHR9; R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually H or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is91MF-363628402Attorney Docket No. 22419-20010.40 individually 1, 2, 3, 4, 5, or 6; R7is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker; R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)pO(CY2)q]tCY2 — X3; R9is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker, with the proviso that exactly one of R7and R9is the point of attachment to the linker; and each X3is individually — H, —OH, — SH, or — NH2.

[0219] In some embodiments, the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein: R1and R2are each individually selected from the group consisting of hydrogen, halogen, -OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O-(Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring; R3is a hydrogen or an unsubstituted Ci-Ce alkyl; R4is — (Ci-Ce alkyl)-X2; X1is — O — ; X2is — OR9; R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0; R7is H; and R9is the point of attachment to the linker. In some embodiments, n4is 1. In some embodiments, n4is 2.

[0220] In some embodiments, the topoisomerase I inhibitor comprises Formula (II) having the structure:92MF-363628402Attorney Docket No. 22419-20010.40wherein: R1is methyl; R2is fluoro; R3is hydrogen; R4is a substituted or an unsubstituted - (C2 alkyl)-X2; and n4is 2, and wherein the remaining variables are as described herein. In some embodiments, L2is absent; n1is the integer 2 or 5; L4is gly-gly-phe-gly; L5is - [NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; and L7is absent, and wherein the remaining variables are as described herein.

[0221] In some embodiments, the topoisomerase I inhibitor comprises Formula (II) having the structure:wherein: R1is methyl; R2is fluoro; R3is hydrogen; R4is a substituted or an unsubstituted - (C2 alkyl)-X2; and n4is 2, and wherein the remaining variables are as described herein.

[0222] In some embodiments, the topoisomerase I inhibitor is represented by:wherein R9is the point of attachment to the linker.

[0223] In some embodiments, the topoisomerase I inhibitor is represented by:93MF-363628402Attorney Docket No. 22419-20010.40wherein R9is the point of attachment to the linker.

[0224] In some embodiments, the topoisomerase I inhibitor is represented by:wherein R9is the point of attachment to the linker.

[0225] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:

[0226] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:94MF-363628402Attorney Docket No. 22419-20010.40

[0227] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:

[0228] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are

[0229] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:95MF-363628402Attorney Docket No. 22419-20010.40

[0230] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are

[0231] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:96MF-363628402Attorney Docket No. 22419-20010.40

[0232] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:

[0233] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:97MF-363628402Attorney Docket No. 22419-20010.40

[0234] In some embodiments, the topoisomerase I inhibitor and the linker, taken together, are represented by:

[0235] In some embodiments, the topoisomerase I inhibitor is exatecan. In some embodiments the topoisomerase I inhibitor is an exatecan derivative. In some embodiments the topoisomerase I inhibitor is selected from the group consisting of exatecan, deruxtecan (i.e., DXd), SN-38, belotecan, topotecan, adizutecan, YL0014, tirumotecan, rezetecan, and A- 1743332. In some embodiments, the topoisomerase I inhibitor is deruxtecan. In some embodiments, the topoisomerase I inhibitor is SN-38. In some embodiments, the topoisomerase I inhibitor is belotecan. In some embodiments, the topoisomerase I inhibitor is topotecan. In some embodiments, the topoisomerase I inhibitor is adizutecan. In some embodiments, the topoisomerase I inhibitor is YL0014. In some embodiments, the topoisomerase I inhibitor is tirumotecan. In some embodiments, the topoisomerase I inhibitor is rezetecan. In some embodiments, the topoisomerase I inhibitor is A- 1743332.98MF-363628402Attorney Docket No. 22419-20010.40

[0236] In some embodiments, the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 11. In some embodiments, the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 11. In some embodiments, the VL comprises an amino acid sequence according to SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the antibody comprises a Fc domain comprising one or more modifications that reduces binding to an Fc receptor, reduces binding to complement component Cl q, or both binding to an Fc receptor and reduces binding to complement component Clq. In some embodiments, the antibody comprises a Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA) or L234A / L235A / P329G (LALAPG). In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of less than 10’5M to 1015M. In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’6M to 1010M. In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’9M to 1010M.99MF-363628402Attorney Docket No. 22419-20010.40

[0237] In certain embodiments, provided herein is an immunoconjugate having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein: Ab is an antibody or an antigen-binding fragment thereof that binds NCAM1, comprising: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT);L1isZ1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl; L3is -(CH2)n1-C(=O)- or -(CH2CH20)n1-(CH2)n1C(=0)-; each n1is individually an integer from 0 to 10; L4is a tetrapeptide residue; L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent,100MF-363628402Attorney Docket No. 22419-20010.40D is a drug moiety; and n is an integer from 1 to 10.

[0238] In certain embodiments, provided herein is an immunoconjugate having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein: Ab is an antibody or an antigen-binding fragment thereof that binds NCAM1, comprising: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT);L1isL2is absent,101MF-363628402Attorney Docket No. 22419-20010.40Z1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl; L3is -(CH2)n1-C(=O)- or -(CH2CH20)n1-(CH2)n1C(=0)-; each n1is individually an integer from 0 to 10; L4is a tetrapeptide residue; L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent orL7is absent,D is a drug moiety; and n is an integer from 1 to 10.

[0239] In some embodiments, D in Formula (I) is a drug moiety of Formula (II- A) having the structure:wherein: R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Cehaloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and -[(CY2)pO(CY2)q]tCY3, or a substituted or102MF-363628402Attorney Docket No. 22419-20010.40 unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring; R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-C6haloalkyl, or — [(CY2)pO(CY2)q]tCY3; R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci-C6haloalkynyl)-X2; X1is — O — , — S(O)W— , — NH — , — O — (C=O) — , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ; X2is — OR9, — SR9, or — NHR9; R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually H or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6; R7is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker; R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)PO(CY2)q]tCY2 — X3; R9is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker, with the proviso that exactly one of R7and R9is the point of attachment to the linker; and each X3is individually — H, —OH, — SH, or — NH2.

[0240] In some embodiments, D in Formula (I) is a drug moiety of Formula (II) having the structure:wherein: R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or103MF-363628402Attorney Docket No. 22419-20010.40 unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and -[(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring; R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-C6haloalkyl, or — [(CY2)PO(CY2)q]tCY3; R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci-C6haloalkynyl)-X2; X1is — O — , — S(O)W— , — NH — , — O — (C=O) — , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ; X2is — OR9, — SR9, or — NHR9; R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually H or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6; R7is H, —COR8, — CO2R8, —(CO)— NHR8, L4, L5, L6, or L7; R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)pO(CY2)q]tCY2— X3; R9is H, —COR8, — CO2R8, —(CO)— NHR8, L4, L5, L6, or L7, with the proviso that exactly one of R7and R9is L4, L5, L6, or L7; and each X3is individually — H, — OH, — SH, or — NH2.

[0241] In some embodiments, D in Formula (I) is a drug moiety of Formula (II) having the structure:104MF-363628402Attorney Docket No. 22419-20010.40 wherein: R1and R2are each individually selected from the group consisting of hydrogen, halogen, -OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O-(Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring; R3is a hydrogen or an unsubstituted Ci-Ce alkyl; R4is a substituted or an unsubstituted — (Ci-Ce alkyl)-X2; X1is — O — ; X2is — OR9; R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0; R7is H; and R9is L4, L5, L6, or L7; and wherein the remaining variables are as described herein. In some embodiments, n4is 1. In some embodiments, n4is 2.

[0242] In some embodiments, D in Formula (I) is a drug moiety of Formula (II) having the structure:wherein: R1is methyl; R2is fluoro; R3is hydrogen; R4is a substituted or an unsubstituted - (C2 alkyl)-X2; and n4is 2, and wherein the remaining variables are as described herein.

[0243] In some embodiments, L2is absent; nl is the integer 2 or 5; L4is gly-gly-phe-gly; L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; and L7is absent, and wherein the remaining variables are as described herein.

[0244] In some embodiments, L6is. In some embodiments, L6is105MF-363628402Attorney Docket No. 22419-20010.40

[0245] In some embodiments, L7is absent. In some embodiments,. In some embodiments,

[0246] In some embodiments, D in Formula (I) is a drug moiety of Formula (II) having the structure:wherein: R1is methyl; R2is fluoro; R3is hydrogen; R4is a substituted or an unsubstituted - (C2 alkyl)-X2; and n4is 2, and wherein the remaining variables are as described herein.

[0247] In some embodiments, D in Formula (I) is represented by:106MF-363628402Attorney Docket No. 22419-20010.40

[0248] In some embodiments, D in Formula (I) is represented by:

[0249] In some embodiments, D in Formula (I) is represented by:

[0250] In some embodiments, D in Formula (I) is represented by:107MF-363628402Attorney Docket No. 22419-20010.40

[0251] In some embodiments, Formula (I) is:wherein n is as described herein.

[0252] In some embodiments, Formula (I) is:wherein n is as described herein.

[0253] In some embodiments, Formula (I) is:108MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.

[0254] In some embodiments, Formula (I) is represented by:wherein n is as described herein.

[0255] In some embodiments, Formula (I) is represented by:wherein n is as described herein.

[0256] In some embodiments, Formula (I) is:109MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.

[0257] In some embodiments, Formula (I) is:wherein n is as described herein.

[0258] In some embodiments, Formula (I) is:110MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.

[0259] In some embodiments, Formula (I) is represented by:111MF-363628402Attorney Docket No. 22419-20010.40wherein n is as described herein.wherein n is as described herein.

[0261] The immunoconjugate of Formula (I) described herein comprises an antibody conjugated to a payload-linker, wherein the payload is a topoisomerase I inhibitor as described herein, and n is an is an integer from 1 to 10 and represents the number of topoisomerase I-inhibiting payload-linkers conjugated to an individual antibody in the immunoconjugate.

[0262] In some embodiments, n is an integer from 4 to 8. In some embodiments, n is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, n is an integer up to 8. In some embodiments, n is an integer of 2. In some embodiments, n is an integer of 4. In some embodiments, n is an integer of 8.112MF-363628402Attorney Docket No. 22419-20010.40

[0263] In some embodiments, n is 1. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4. In some embodiments, n is 5. In some embodiments, n is 6. In some embodiments, n is 7. In some embodiments, n is 8. In some embodiments, n is 9. In some embodiments, n is 10.

[0264] In some aspects, provided herein is a composition comprising a plurality of any of the immunoconjugates provided herein. In some embodiments, the average n is a number from 1 to 10. In some embodiments, the average n within a composition comprising the immunoconjugates of Formula (I) is a number from 1 to 10. In some embodiments, the average n is about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7,2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8,4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9,7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0,9.1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, or 10.0.

[0265] For example, in some embodiments, each immunoconjugate of a plurality of immunoconjugates comprises Formula (I), wherein n is a different integer and n is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 (i.e., plurality of immunoconjugates comprising heterogenous DAR or mixed DAR). In some embodiments, at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85% 90%, 95%, or more) of the plurality of immunoconjugates comprise Formula (I), wherein n is 4 and the remaining immunoconjugates comprise Formula (I), wherein n is a different integer and n is an integer of 1, 2, 3, 5, 6, 7, 8, 9, or 10. In some embodiments, at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85% 90%, 95%, or more) of the plurality of immunoconjugates comprise Formula (I), wherein n is 6 and the remaining immunoconjugates comprise Formula (I), wherein n is a different integer and n is an integer of 1, 2, 3, 4, 5, 7, 8, 9, or 10. In some embodiments, at least 50% (e.g., 55%, 60%, 65%, 70%, 75%, 80%, 85% 90%, 95%, or more) of the plurality of immunoconjugates comprise Formula (I), wherein n is 8 and the remaining immunoconjugates comprise Formula (I), wherein n is a different integer and n is an integer of 1, 2, 3, 4, 5, 6, 7, 9, or 10.

[0266] In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 1. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 2. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 3. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 4. In some embodiments, the average n within a pharmaceutical composition comprising the113MF-363628402Attorney Docket No. 22419-20010.40 immunoconjugates of Formula (I) is about 5. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 6. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 7. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 8. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 9. In some embodiments, the average n within a pharmaceutical composition comprising the immunoconjugates of Formula (I) is about 10.

[0267] In some embodiments, the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 11. In some embodiments, the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 11. In some embodiments, the VL comprises an amino acid sequence according to SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the antibody comprises a Fc domain comprising one or more modifications that reduces binding to an Fc receptor, reduces binding114MF-363628402Attorney Docket No. 22419-20010.40 to complement component Clq, or both binding to an Fc receptor and reduces binding to complement component Clq. In some embodiments, the antibody comprises a Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA) or L234A / L235A / P329G (LALAPG). In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of less than IO’5M to 1015M. In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’6M to 1010M. In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’9M to 1010M.

[0268] In certain embodiments, also provided herein is an immunoconjugate having Formulawherein: Ab is an antibody or an antigen-binding fragment thereof that binds NCAM1, comprising: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT);S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;115MF-363628402Attorney Docket No. 22419-20010.40Z1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl; L3is -(CH2)n1-C(=O)- or -(CH2CH20)n1-(CH2)n1C(=0)-; each n1is individually an integer from 0 to 10; L4is a tetrapeptide residue; L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent,wherein D has the structure:and n is an integer from 1 to 10.

[0269] In certain embodiments, also provided herein is an immunoconjugate having Formula(I),Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein: Ab is an antibody or an antigen-binding fragment thereof that binds NCAM1, comprising: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to (SFGMH) SEQ ID116MF-363628402Attorney Docket No. 22419-20010.40NO: 1; (ii) a CDR2 having an amino acid sequence according to (YISSGSFTIYYADSVKG) SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to (MRKGYAMDY) SEQ ID NO: 3; and a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to (RSSQIIIHSDGNTYLE) SEQ ID NO: 4; (ii) a CDR2 having an amino acid sequence according to (KVSNRFS) SEQ ID NO: 5; and (iii) a CDR3 having an amino acid sequence according to (FQGSHVPHT) SEQ ID NO: 6;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof; L1isL2is absent,Z1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl; L3is -(CH2)n1-C(=O)- or -(CH2CH20)n1-(CH2)n1C(=0)-; each n1is individually an integer from 0 to 10; L4is a tetrapeptide residue; L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent orL7is absent,wherein D has the structure:117MF-363628402Attorney Docket No. 22419-20010.40and n is an integer from 1 to 10.

[0270] In some embodiments, L6isIn some embodiments, L6is118MF-363628402Attorney Docket No. 22419-20010.40

[0271] In some embodiments, L7is absent. In some embodiments,

[0272] In some embodiments, n is an integer from 4 to 8. In some embodiments, n is an integer up to 8. In some embodiments, n is an integer of 2. In some embodiments, n is an integer of 4. In some embodiments, n is an integer of 8. In some embodiments, the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 11. In some embodiments, the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7, SEQ ID NO: 8, or SEQ ID NO: 11. In some embodiments, the VL comprises an amino acid sequence according to SEQ ID NO: 9, SEQ ID NO: 10, or SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 10. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9. In some embodiments, the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.119MF-363628402Attorney Docket No. 22419-20010.40

[0273] In some embodiments, provided herein is an immunoconjugate having Formula (I),Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a heavy chain variable region (VH) comprising the amino acid sequence according to SEQ IDNO: 7; and a light chain variable region (VL) comprising the amino acid sequence according to SEQ ID NO: 10;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L2is absent;L3is -(CH2)n1-C(=O)-; n1is the integer 2;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; andL6is absent;L7is absent;D has the structure:120MF-363628402Attorney Docket No. 22419-20010.40 and n is 8.

[0274] In some embodiments, provided herein is an immunoconjugate having Formula (I),Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a heavy chain variable region (VH) comprising the amino acid sequence according to SEQ IDNO: 7; and a light chain variable region (VL) comprising the amino acid sequence according to SEQ ID NO: 10;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L2is absent;L3is -(CH2)n1-C(=O)-; n1is the integer 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; andL6is absent;L7is absent;D has the structure:121MF-363628402Attorney Docket No. 22419-20010.40and n is 8.

[0275] In some embodiments, the antibody comprises a Fc domain comprising one or more modifications that reduces binding to an Fc receptor, reduces binding to complement component Cl q, or both binding to an Fc receptor and reduces binding to complement component Clq. In some embodiments, the antibody comprises a Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0276] In some embodiments, the payload-linkers described herein may be conjugated to the anti-NCAMl antibodies via cysteine modifications to allow for site-specific conjugation. In some embodiments, the anti-NCAMl antibodies described herein comprise one or more cysteine modifications in the HC and / or LC to allow for one or more site-specific conjugations. In some embodiments, the LC comprises one or more mutations, including but not limited to S114C, S121C, S127C, A153C, N158C, and V205C, wherein the positions are based on EU numbering. In some embodiments, the HC comprises one or more mutations, including but not limited to S239C, V282C, S324C, and S375C, wherein the positions are based on EU numbering.

[0277] In some embodiments, the immunoconjugates comprising anti-NCAMl antibodies described herein comprise humanized anti-NCAMl antibodies or antigen-binding fragments thereof.

[0278] In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of less than 10’4M to less than 1015M.

[0279] In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’6M to 1010M.

[0280] In some embodiments, the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’9M to 1010M.122MF-363628402Attorney Docket No. 22419-20010.40

[0281] In some embodiments, a linker described herein is an enzymatically cleavable linker. In some embodiments, the enzymatically cleavable linker is a B-glucuronic acid-based linker. Facile release of the drug may be realized through cleavage of the B-glucuronide glycosidic bond by the lysosomal enzyme B-glucuronidase. This enzyme is present abundantly within lysosomes and is overexpressed in some tumor types, while the enzyme activity outside cells is low. B-Glucuronic acid-based linkers may be used to circumvent the tendency of an ADC to undergo aggregation due to the hydrophilic nature of B-glucuronides. In some embodiments, B-glucuronic acid-based linkers are linkers for ADCs linked to hydrophobic drugs.TREATMENT OF CANCER

[0282] Described herein, in certain embodiments, are methods of treating a cancer in a patient in need thereof, the method comprising administering to the patient an effective amount of anti-NCAMl antibodies or immunoconjugates (e.g., an antibody-drug conjugate).

[0283] In some embodiments, the disclosure additionally provides methods of identifying subjects who are candidates for treatment with an anti-NCAMl antibody or immunoconjugate having NCAM1 -targeting effects. Thus, in some embodiments, the disclosure provides a method of identifying a patient who can benefit from treatment with an anti-NCAMl antibody or immunoconjugate disclosed herein. In some embodiments, the patient has a tumor. In some embodiments, the tumor is a solid tumor. In some embodiments, the tumor sample is from a primary tumor. In alternative embodiments, the tumor sample is a metastatic lesion. In some embodiments, the patient has a tumor that expresses NCAM1. Binding of antibody or immunoconjugate to NCAMl-expressing tumor cells through a binding interaction NCAM1 can be measured using any assay, such as immunohistochemistry or flow cytometry. In some embodiments, binding of antibody to at least 0.2%, at least 0.5%, at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, or at least 50% of the NCAMl-expressing tumor cells in a sample may be used as a selection criterion for determining a patient to be treated with an anti-NCAMl antibody immunoconjugate as described herein.

[0284] An anti-NCAMl antibody or an anti-NCAMl antibody immunoconjugate disclosed herein can be used to treat several different cancers. In some embodiments, a cancer patient who can benefit from the treatment of the anti-NCAMl antibody or antibody immunoconjugate (e.g., an anti-NCAMl antibody-drug conjugate) has a cancer expressing a NCAM1.123MF-363628402Attorney Docket No. 22419-20010.40

[0285] In some embodiments, the cancer is a neuroendocrine cancer. In some embodiments, the cancer is a non-squamous carcinoma. In some embodiments, the cancer is multiple myeloma, glioblastoma, sarcoma, non-small cell lung cancer, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, small intestinal neuroendocrine tumor, or renal cell carcinoma. In some embodiments, the cancer is multiple myeloma. In some embodiments, the cancer is glioblastoma. In some embodiments, the cancer is sarcoma. In some embodiments, the cancer is non-small cell lung cancer (NSCLC; e.g., NSCLC- adenocarcinoma (NSCLC-AD) or NSCLC-squamous (NSCLC-SQ)). In some embodiments, the cancer is small cell lung cancer. In some embodiments, the cancer is Merkel cell carcinoma. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is large cell neuroendocrine carcinoma. In some embodiments, the cancer is neuroendocrine prostate cancer. In some embodiments, the cancer is pancreatic neuroendocrine tumor. In some embodiments, the cancer is neuroblastoma. In some embodiments, the cancer is small intestinal neuroendocrine tumor. In some embodiments, the cancer is renal cell carcinoma.

[0286] In some embodiments, methods of the disclosure comprise administering an anti- NCAM1 antibody or immunoconjugate disclosed herein, or a variant thereof, as a pharmaceutical composition to a cancer patient in a therapeutically effective amount using a dosing regimen suitable for treatment of the cancer. The composition can be formulated for use in a variety of drug delivery systems. One or more physiologically acceptable excipients or carriers can also be included in the compositions for proper formulation.

[0287] The anti-NCAMl antibody or immunoconjugate is provided in a solution suitable for administration to the patient, such as a sterile isotonic aqueous solution for injection. The antibody is dissolved or suspended at a suitable concentration in an acceptable carrier. In some embodiments the carrier is aqueous, e.g., water, saline, phosphate buffered saline, and the like. The compositions may contain auxiliary pharmaceutical substances as required to approximate physiological conditions, such as pH adjusting and buffering agents, tonicity adjusting agents, and the like.ADMINISTRATION

[0288] The pharmaceutical compositions are administered to a patient in an amount sufficient to cure or at least partially arrest the disease or symptoms of the disease and its complications. An amount adequate to accomplish this is defined as a “therapeutically124MF-363628402Attorney Docket No. 22419-20010.40 effective dose.” A therapeutically effective dose is determined by monitoring a patient’s response to therapy. Typical benchmarks indicative of a therapeutically effective dose includes the amelioration of symptoms of the disease in the patient. Amounts effective for this use will depend upon the severity of the disease and the general state of the patient’ s health, including other factors such as age, weight, gender, administration route, and the like Single or multiple administrations of the antibody or immunoconjugate may be administered depending on the dosage and frequency as required and tolerated by the patient. In any event, the methods provide a sufficient quantity of NCAM1 -targeting antibody or immunoconjugate to effectively treat the patient.

[0289] An anti-NCAMl antibody or immunoconjugate can be administered by any suitable means, including, for example, parenteral, intrapulmonary, and intranasal, administration, as well as local administration, such as intratumor administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0290] The dose of antibody or immunoconjugate is chosen to provide effective therapy for the subject.PHARMACEUTICAL COMPOSITIONS

[0291] The present disclosure also features pharmaceutical compositions that contain a therapeutically effective amount of the anti-NCAMl antibody or immunoconjugate described herein. The composition can be formulated for use in a variety of drug delivery systems. One or more physiologically acceptable excipients or carriers can also be included in the composition for proper formulation. Suitable formulations for use in the present disclosure are found in Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, Pa., 17th ed., 1985. For a brief review of methods for drug delivery, see, e.g., Langer (Science 249:1527-1533, 1990).

[0292] In some embodiments, a pharmaceutical composition may contain formulation materials for modifying, maintaining or preserving, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, adsorption or penetration of the composition. In such embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants (such as ascorbic acid, sodium sulfite or sodium hydrogen- sulfite); buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates or other organic acids); bulking agents (such as mannitol or glycine); chelating agents (such as ethylenediamine tetraacetic acid (EDTA)); complexing agents (such as caffeine,125MF-363628402Attorney Docket No. 22419-20010.40 polyvinylpyrrolidone, beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose or dextrins); proteins (such as serum albumin, gelatin or immunoglobulins); coloring, flavoring and diluting agents; emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight polypeptides; salt-forming counterions (such as sodium); preservatives (such as benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide); solvents (such as glycerin, propylene glycol or polyethylene glycol); sugar alcohols (such as mannitol or sorbitol); suspending agents; surfactants or wetting agents (such as pluronics, PEG, sorbitan esters, polysorbates such as polysorbate 20, polysorbate, triton, tromethamine, lecithin, cholesterol, tyloxapal); stability enhancing agents (such as sucrose or sorbitol); tonicity enhancing agents (such as alkali metal halides, preferably sodium or potassium chloride, mannitol sorbitol); delivery vehicles; diluents; excipients and / or pharmaceutical adjuvants (see, Remington’s Pharmaceutical Sciences, 18th ed. (Mack Publishing Company, 1990)).

[0293] In some embodiments, a pharmaceutical composition may contain nanoparticles, e.g., polymeric nanoparticles, liposomes, or micelles.

[0294] In some embodiments, a pharmaceutical composition may contain a sustained- or controlled-delivery formulation. Techniques for formulating sustained- or controlled-delivery means, such as liposome carriers, bio-erodible microparticles or porous beads and depot injections, are also known to those skilled in the art. Sustained-release preparations may include, e.g., porous polymeric microparticles or semipermeable polymer matrices in the form of shaped articles, e.g., films, or microcapsules. Sustained release matrices may include polyesters, hydrogels, polylactides, copolymers of L-glutamic acid and gamma ethyl-L- glutamate, poly (2-hydroxyethyl-inethacrylate), ethylene vinyl acetate, or poly-D(-)-3- hydroxybutyric acid. Sustained release compositions may also include liposomes that can be prepared by any of several methods known in the art.

[0295] Pharmaceutical compositions containing an anti-NCAMl antibody or immunoconjugate disclosed herein can be presented in a dosage unit form and can be prepared by any suitable method. A pharmaceutical composition should be formulated to be compatible with its intended route of administration. Examples of routes of administration are intravenous (IV), intradermal, inhalation, transdermal, topical, transmucosal, intrathecal and rectal administration.126MF-363628402Attorney Docket No. 22419-20010.40

[0296] Useful formulations can be prepared by methods known in the pharmaceutical art. For example, see Remington’s Pharmaceutical Sciences, 18th ed. (Mack Publishing Company, 1990). Formulation components suitable for parenteral administration include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as EDTA; buffers such as acetates, citrates or phosphates; and agents for the adjustment of tonicity such as sodium chloride or dextrose.

[0297] For intravenous or subcutaneous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor ELTM (BASF, Parsippany, NJ) or phosphate buffered saline (PBS). The carrier should be stable under the conditions of manufacture and storage, and should be preserved against microorganisms. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyetheylene glycol), and suitable mixtures thereof.

[0298] An intravenous or subcutaneous drug delivery formulation may be contained in a syringe, pen, or bag. In some embodiments, the bag is connected to a channel comprising a tube and / or a needle. In some embodiments, the formulation is a lyophilized formulation or a liquid formulation.

[0299] These compositions may be sterilized by conventional sterilization techniques, or may be sterile filtered. The resulting aqueous solutions may be packaged for use as-is, or lyophilized, the lyophilized preparation being combined with a sterile aqueous carrier prior to administration.

[0300] A polyol, which acts as a tonicifier and may stabilize the anti-NCAMl antibody or immunoconjugate, may also be included in the formulation. The polyol is added to the formulation in an amount which may vary with respect to the desired isotonicity of the formulation. In some embodiments, the aqueous formulation is isotonic. The amount of polyol added may also be altered with respect to the molecular weight of the polyol. For example, a lower amount of a monosaccharide (e.g., mannitol) is added, compared to a disaccharide (such as trehalose). In some embodiments, the polyol which is used in the formulation as a tonicity agent is mannitol.

[0301] A detergent or surfactant may also be added to the formulation. Exemplary detergents include nonionic detergents such as polysorbates (e.g., polysorbates 20, 80 etc.) or poloxamers (e.g., poloxamer 188). The amount of detergent added is such that it reduces aggregation of the formulated antibody and / or minimizes the formation of particulates in the 127MF-363628402Attorney Docket No. 22419-20010.40 formulation and / or reduces adsorption. In some embodiments, the formulation may include a surfactant which is a polysorbate. In some embodiments, the formulation may contain the detergent polysorbate 80 or Tween 80. Tween 80 is a term used to describe polyoxyethylene (20) sorbitanmonooleate (see Fiedler, Lexikon der Hifsstoffe, Editio Cantor Verlag Aulendorf, 4th edi., 1996).

[0302] In embodiments, the protein product of the present disclosure is formulated as a liquid formulation. In some embodiments, the liquid formulation is prepared in combination with a sugar at stabilizing levels. In some embodiments, the liquid formulation is prepared in an aqueous carrier. In some embodiments, a stabilizer is added in an amount no greater than that which may result in a viscosity undesirable or unsuitable for intravenous administration. In some embodiments, the sugar is disaccharides, e.g., sucrose. In some embodiments, the liquid formulation may also include one or more of a buffering agent, a surfactant, and a preservative.

[0303] In some embodiments, the pH of the liquid formulation is set by addition of a pharmaceutically acceptable acid and / or base. In some embodiments, the pharmaceutically acceptable acid is hydrochloric acid. In some embodiments, the base is sodium hydroxide.

[0304] The aqueous carrier of interest herein is one which is pharmaceutically acceptable (safe and non-toxic for administration to a human) and is useful for the preparation of a liquid formulation. Illustrative carriers include sterile water for injection (SWFI), bacteriostatic water for injection (BWFI), a pH buffered solution (e.g., phosphate-buffered saline), sterile saline solution, Ringer's solution or dextrose solution.

[0305] A preservative may be optionally added to the formulations herein to reduce bacterial action. The addition of a preservative may, for example, facilitate the production of a multiuse (multiple-dose) formulation.

[0306] The anti-NCAMl antibody or immunoconjugate may be lyophilized to produce a lyophilized formulation including the proteins and a lyoprotectant. The lyoprotectant may be sugar, e.g., disaccharides. In some embodiments, the lyoprotectant is sucrose or maltose. The lyophilized formulation may also include one or more of a buffering agent, a surfactant, a bulking agent, and / or a preservative.

[0307] The amount of sucrose or maltose useful for stabilization of the lyophilized drug product may be in a weight ratio of at least 1:2 protein to sucrose or maltose. In some embodiments, the protein to sucrose or maltose weight ratio is of from 1:2 to 1:5. In some embodiments, the pH of the formulation, prior to lyophilization, is set by addition of a pharmaceutically acceptable acid and / or base. In some embodiments, the pharmaceutically 128MF-363628402Attorney Docket No. 22419-20010.40 acceptable acid is hydrochloric acid. In some embodiments, the pharmaceutically acceptable base is sodium hydroxide.

[0308] A patient’s dose can be tailored to the approximate body weight or surface area of the patient. Other factors in determining the appropriate dosage can include the disease or condition to be treated or prevented, the severity of the disease, the route of administration, and the age, sex, and medical condition of the patient. Further refinement of the calculations necessary to determine the appropriate dosage for treatment is routinely made by those skilled in the art, especially in light of the dosage information and assays disclosed herein. The dosage can also be determined through the use of known assays for determining dosages used in conjunction with appropriate dose-response data. An individual patient's dosage can be adjusted as the progress of the disease is monitored. Blood levels of the targetable construct or complex in a patient can be measured to see if the dosage needs to be adjusted to reach or maintain an effective concentration. Pharmacogenomics may be used to determine which targetable constructs and / or complexes, and dosages thereof, are most likely to be effective for a given individual (Schmitz et al., Clinica Chimica Acta 308: 43-53, 2001;Steimer et al., Clinica Chimica Acta 308: 33-41, 2001).

[0309] In some embodiments, the antibody may be stored at 10 mg / ml or 20 mg / ml in a sterile isotonic aqueous solution. The solution can comprise agents such as buffering agents and stabilizing agents. For example, in some embodiments, a buffering agent such as histidine is included to maintain a formulation pH of about 5.5. Additional reagents such as sucrose or alternatives can be added to prevent aggregation and fragmentation in solution and during freezing and thawing. Agents such as polysorbate 80 or an alternative can be included to lower surface tension and stabilizes the antibody against agitation-induced denaturation and air-liquid and ice-liquid surface denaturation. In some embodiments, the solution for injection is stored at 4°C and is diluted in either 100 ml or 200 ml 0.9% sodium chloride for injection prior to administration to the patient.EXEMPLARY EMBODIMENTS

[0310] The following exemplary embodiments are provided herein:

[0311] Embodiment 1. An antibody or antigen-binding fragment thereof that binds NCAM1, comprising: a) a heavy chain variable region (VH) comprising 96% sequence identity to SEQ ID NO: 7 or 99% sequence identity SEQ ID NO: 8; and129MF-363628402Attorney Docket No. 22419-20010.40 b) a light chain variable region (VL) comprising 100% sequence identity to SEQ ID NO: 9 or 90% sequence identity to SEQ ID NO: 10.

[0312] Embodiment 2. An antibody or antigen-binding fragment thereof that binds NCAM1, comprising: a) a heavy chain variable region (VH) comprising 100% sequence identity to SEQ ID NO: 11; and b) a light chain variable region (VL) comprising 100% sequence identity to SEQ ID NO: 9 or 90% sequence identity to SEQ ID NO: 10.

[0313] Embodiment 3. An antibody or antigen-binding fragment thereof that binds NCAM1, comprising: a) a heavy chain variable region (VH) comprising 96% sequence identity to SEQ IDNO: 7 or 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising 100% sequence identity to SEQ ID NO: 12.

[0314] Embodiment 4. An antibody or antigen-binding fragment thereof that binds NCAM1, comprising: a) a heavy chain variable region (VH) comprising 96% sequence identity to SEQ ID NO: 7 or 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO: 12, and wherein: i) the VL comprises a substitution at amino acid position 79 (sequential), numbering according to the sequence of SEQ ID NO: 12; ii) the VL comprises a substitution at amino acid position 82 (sequential), numbering according to the sequence of SEQ ID NO: 12; iii) the VL comprises substitutions at amino acid positions 79 and 82 (sequential), numbering according to the sequence of SEQ ID NO: 12; iv) the VL comprises a substitution at amino acid position 74 (Kabat), numbering according to the sequence of SEQ ID NO: 12; v) the VL comprises a substitution at amino acid position 77 (Kabat), numbering according to the sequence of SEQ ID NO: 12; or vi) the VL comprises substitutions at amino acid positions 74 and 77 (Kabat), numbering according to the sequence of SEQ ID NO: 12.

[0315] Embodiment 5. An immunoconjugate, comprising:130MF-363628402Attorney Docket No. 22419-20010.40 a) an antibody or antigen-binding fragment thereof that binds to NCAM1, wherein the antibody comprises: i) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and ii) a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); and b) a topoisomerase I inhibitor conjugated to the antibody or the antigen-binding fragment thereof using a linker.

[0316] Embodiment 6. The immunoconjugate of embodiment 5, wherein the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, - CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Cehaloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and -[(CY2)PO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;131MF-363628402Attorney Docket No. 22419-20010.40R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)pO(CY2)q]tCY3;R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-C6 haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci-Ce haloalkynyl)-X2;X1is — O— , — S(O)W— , — NH— , — O— (C=O)— , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ;X2is —OR9, —SR9, or — NHR9;R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)O(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually H or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6;R7is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker;R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)pO(CY.2)q]tCY2— X3;R9is H, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker, with the proviso that exactly one of R7and R9is the point of attachment to the linker; and each X3is individually — H, — OH, — SH, or — NH2.

[0317] Embodiment 7. The immunoconjugate of embodiment 6, wherein the topoisomerase I inhibitor is represented by Formula (II) having the structure:132MF-363628402Attorney Docket No. 22419-20010.40wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, - OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O- (Ci-C6alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or an unsubstituted Ci-Ce alkyl;R4is (Ci-C6alkyl)-X2;X1is — O— ;X2is —OR9;R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0;R7is H; andR9is the point of attachment to the linker.

[0318] Embodiment 8. The immunoconjugate of embodiment 7, wherein n4is 1.

[0319] Embodiment 9. The immunoconjugate of embodiment 7, wherein n4is 2.

[0320] Embodiment 10. The immunoconjugate of embodiment 5, wherein the topoisomerase I inhibitor comprises Formula (II) having the structure:133MF-363628402Attorney Docket No. 22419-20010.40R1is methyl;R2is fluoro;R3is hydrogen;R4is an unsubstituted -(C2 alkyl)-X2; and n4is 2.

[0321] Embodiment 11. The immunoconjugate of embodiment 5, wherein:L2is absent; n1is the integer 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; andL7is absent.

[0322] Embodiment 12. The immunoconjugate of embodiment 7, wherein the topoisomeraseI inhibitor comprises Formula (II) having the structure:wherein:R1is methyl;R2is fluoro;R3is hydrogen;134MF-363628402Attorney Docket No. 22419-20010.40R4is an unsubstituted -(C2 alkyl)-X2; and n4is 2.

[0323] Embodiment 13. The immunoconjugate of embodiment 7, wherein the topoisomeraseI inhibitor is represented by:wherein R9is the point of attachment to the linker.

[0324] Embodiment 14. The immunoconjugate of embodiment 7, wherein the topoisomeraseI inhibitor is represented by:wherein R9is the point of attachment to the linker.

[0325] Embodiment 15. The immunoconjugate of embodiment 7, wherein the topoisomeraseI inhibitor is represented by:135MF-363628402Attorney Docket No. 22419-20010.40 wherein R9is the point of attachment to the linker.

[0326] Embodiment 16. The immunoconjugate of embodiment 5, wherein the topoisomeraseI inhibitor and the linker, taken together, are represented by:

[0327] Embodiment 17. The immunoconjugate of embodiment 5, wherein the topoisomeraseI inhibitor and the linker, taken together, are represented by:

[0328] Embodiment 18. The immunoconjugate of embodiment 5, wherein the topoisomeraseI inhibitor and the linker, taken together, are represented by:136MF-363628402Attorney Docket No. 22419-20010.40

[0329] Embodiment 19. The immunoconjugate of embodiment 5, wherein the topoisomeraseI inhibitor and the linker, taken together, are represented by:

[0330] Embodiment 20. The immunoconjugate of embodiment 5, wherein the topoisomeraseI inhibitor and the linker, taken together, are represented by:137MF-363628402Attorney Docket No. 22419-20010.40

[0331] Embodiment 21. An immunoconjugate having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n (I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds NCAM1, comprising: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT);L1isL2is absent,138MF-363628402Attorney Docket No. 22419-20010.40Z1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl;E3is -(CH2)n1-C(=O)- or -(CH2CH2O)n1-(CH2)n1C(=O)-;L4is a tetrapeptide residue;L5is absent or -[NH(CH2)n2]n3; n2is an integer of 0 to 6; n3is an integer from 0 to 2;L6is absent,D is a drug moiety; and n is an integer from 1 to 10.

[0332] Embodiment 22. The immunoconjugate of embodiment 21, wherein D in Formula (I) is a drug moiety of Formula (II) having the structure:139MF-363628402Attorney Docket No. 22419-20010.40wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, - CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and -[(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3;R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-C6 haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-C6alkynyl)-X2, or a substituted or an unsubstituted — (Ci-Ce haloalkynyl)-X2;X1is — O— , — S(O)W— , — NH— , — O— (C=O)— , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ;X2is —OR9, —SR9, or — NHR9;R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)O(CY2)q]tCY.3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2;140MF-363628402Attorney Docket No. 22419-20010.40 each Y is individually H or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6;R7is H, —COR8, — CO2R8, — (CO)— NHR8, L4, L5, L6, or L7;R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)pO(CY.2)q]tCY2— X3;R9is H, — COR8, — CO2R8, — (CO) — NHR8, L4, L5, L6, or L7, with the proviso that exactly one of R7and R9is L4, L5, L6, or L7; and each X3is individually — H, — OH, — SH, or — NH2.

[0333] Embodiment 23. The immunoconjugate of embodiment 22, wherein D in Formula (I) is a drug moiety of Formula (II) having the structure:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, - OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O- (Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or an unsubstituted Ci-Ce alkyl;R4is (Ci-C6alkyl)-X2;X1is — O— ;141MF-363628402Attorney Docket No. 22419-20010.40X2is —OR9;R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0;R7is H; andR9is L4, L5, L6, or L7;

[0334] Embodiment 24. The immunoconjugate of embodiment 23, wherein n4is 1.

[0335] Embodiment 25. The immunoconjugate of embodiment 23, wherein n4is 2.

[0336] Embodiment 26. The immunoconjugate of embodiment 22, wherein D in Formula (I) is a drug moiety of Formula (II) having the structure:wherein:R1is methyl;R2is fluoro;R3is hydrogen;R4is an unsubstituted -(C2 alkyl)-X2; and n4is 2.

[0337] Embodiment 27. The immunoconjugate of embodiment 22, wherein:L2is absent; n1is the integer 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; andL7is absent.142MF-363628402Attorney Docket No. 22419-20010.40

[0338] Embodiment 28. The immunoconjugate of embodiment 27, wherein D in Formula (I) is a drug moiety of Formula (II) having the structure:wherein:R1is methyl;R2is fluoro;R3is hydrogen;R4is an unsubstituted -(C2 alkyl)-X2; and n4is 2.

[0339] Embodiment 29. The immunoconjugate of embodiment 23 wherein D in Formula (I) is represented by:wherein R9is E4, E5, L6, or L7.

[0340] Embodiment 30. The immunoconjugate of embodiment 23, wherein D in Formula (I) is represented by:143MF-363628402Attorney Docket No. 22419-20010.40

[0341] Embodiment 31. The immunoconjugate of embodiment 23, wherein D in Formula (I) is represented by:

[0342] Embodiment 32. The immunoconjugate of embodiment 21, wherein Formula (I) is:or144MF-363628402Attorney Docket No. 22419-20010.40

[0343] Embodiment 33. The immunoconjugate of embodiment 21, wherein Formula (I) is:

[0344] Embodiment 34. The immunoconjugate of embodiment 21, wherein Formula (I) is:145MF-363628402Attorney Docket No. 22419-20010.40

[0345] Embodiment 35. The immunoconjugate of embodiment 21, wherein Formula (I) is represented by:146MF-363628402Attorney Docket No. 22419-20010.40

[0346] Embodiment 36. The immunoconjugate of embodiment 21, wherein Formula (I) is represented by:

[0347] Embodiment 37. An immunoconjugate having Formula (I), Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n (I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds NCAM1, comprising: a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to (SFGMH) SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to (YISSGSFTIYYADSVKG) SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to (MRKGYAMDY) SEQ ID NO: 3; and147MF-363628402Attorney Docket No. 22419-20010.40 a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to (RSSQIIIHSDGNTYLE) SEQ ID NO: 4; (ii) a CDR2 having an amino acid sequence according to (KVSNRFS) SEQ ID NO: 5; and (iii) a CDR3 having an amino acid sequence according to (FQGSHVPHT) SEQ ID NO: 6; L1isZ1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl;L3is -(CH2)n1-C(=O)- or -(CH2CH2O)n1-(CH2)n1C(=O)-;L4is a tetrapeptide residue;L5is absent or -[NH(CH2)n2]n3; n2is an integer of 0 to 6; n3is an integer from 0 to 2;L6is absent,wherein D has the structure:148MF-363628402Attorney Docket No. 22419-20010.40and n is an integer from 1 to 10.

[0348] Embodiment 38. The immunoconjugate of any one of embodiments 21-37, wherein n is an integer from 4 to 8.

[0349] Embodiment 39. The immunoconjugate of any one of embodiments 5-38, wherein the VH comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 7 or SEQ ID NO: 8.

[0350] Embodiment 40. The immunoconjugate of any one of embodiments 5-38, wherein the VL comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 9 or SEQ ID NO: 10.

[0351] Embodiment 41. The antibody of embodiment 1 or 4 or the immunoconjugate of any one of embodiments 5-40, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 or SEQ ID NO: 8.

[0352] Embodiment 42. The antibody of embodiment 1 or 4 or the immunoconjugate of any one of embodiments 5-41, wherein the VL comprises an amino acid sequence according to SEQ ID NO: 9 or SEQ ID NO: 10.

[0353] Embodiment 43. The antibody of embodiment 1 or 4 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

[0354] Embodiment 44. The antibody of embodiment 1 or 4 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

[0355] Embodiment 45. The antibody of embodiment 1 or 4 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.149MF-363628402Attorney Docket No. 22419-20010.40

[0356] Embodiment 46. The antibody of embodiment 1 or 4 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

[0357] Embodiment 47. The antibody of embodiment 2 or 3 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

[0358] Embodiment 48. The antibody of embodiment 2 or 3 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

[0359] Embodiment 49. The antibody of embodiment 2 or 3 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 12.

[0360] Embodiment 50. The antibody of embodiment 2 or 3 or the immunoconjugate of any one of embodiments 5-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 12.

[0361] Embodiment 51. The antibody of any one of embodiments 1-4 or the immunoconjugate of any one of embodiments 5-50, wherein the antibody comprises a Fc domain comprising one or more modifications that reduces binding to an Fc receptor, reduces binding to complement component Cl q, or both binding to an Fc receptor and reduces binding to complement component Cl q.

[0362] Embodiment 52. The antibody of any one of embodiments 1-4 or the immunoconjugate of any one of embodiments 5-51, wherein the antibody comprises a Fc domain comprising amino acid modifications L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

[0363] Embodiment 53. The antibody of any one of embodiments 1-4 or the immunoconjugate of any one of embodiments 5-52, wherein the antibody or antibody fragment binds to human NCAM1 with an affinity of 10’7M to 1010M.

[0364] Embodiment 54. A polynucleotide encoding the antibody or the immunoconjugate of any one of embodiments 1-53.

[0365] Embodiment 55. An expression vector comprising a polynucleotide of embodiment 54.

[0366] Embodiment 56. A cell comprising an expression vector of embodiment 55.

[0367] Embodiment 57. A cell capable of producing the antibody or the immunoconjugate of any one of embodiments 1-53.150MF-363628402Attorney Docket No. 22419-20010.40

[0368] Embodiment 58. A pharmaceutical composition comprising the antibody or the immunoconjugate of any one of embodiments 1-53, and a pharmaceutically acceptable excipient, salt, carrier, or diluent.

[0369] Embodiment 59. The pharmaceutical composition of embodiment 58 for use in treating cancer in a subject.

[0370] Embodiment 60. A method of treating cancer compromising administering to a subject in need thereof, a therapeutically effective amount of the antibody or the immunoconjugate of any one of embodiments 1-53.

[0371] Embodiment 61. The method of embodiment 60, further comprising administering an additional anti-cancer therapy selected from surgery, chemotherapy, radiotherapy, and immunotherapy.

[0372] Embodiment 62. The method of embodiment 60, further comprising administering to the subject an immune-modulator, activated lymphocyte cell, kinase inhibitor, or a chemotherapeutic agent.

[0373] Embodiment 63. The method of embodiment 60, wherein the cancer is a solid cancer.

[0374] Embodiment 64. The method of embodiment 60, wherein the cancer is a neuroendocrine cancer.

[0375] Embodiment 65. The method of embodiment 60, wherein the cancer is multiple myeloma, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, or small intestinal neuroendocrine tumor.

[0376] Embodiment 66. The method of embodiment 60, wherein the cancer is NCAM1 positive.

[0377] Embodiment 67. A method of diagnosing or prognosing cancer in a subject, the method comprises determining the expression level of NCAM1 in a biological samples using the antibody or the immunoconjugate of any one of embodiments 1-53.

[0378] Embodiment 68. The use of an effective amount of the antibody or the immunoconjugate of any one of embodiments 1-53 for treating cancer.

[0379] Embodiment 69. The use of embodiment 68, wherein the cancer is a solid cancer.

[0380] Embodiment 70. The use of embodiment 68, wherein the cancer is a neuroendocrine cancer.

[0381] Embodiment 71. The use of embodiment 68, wherein the cancer is multiple myeloma, glioblastoma, sarcoma, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large151MF-363628402Attorney Docket No. 22419-20010.40 cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, or small intestinal neuroendocrine tumor.

[0382] Embodiment 72. A kit comprising the antibody or the immunoconjugate of any one of embodiments 1-53 and instructions for use.EXAMPLES

[0383] The disclosure now being generally described, will be more readily understood by reference to the following examples, which are included merely for purposes of illustration of certain aspects and embodiment of the present disclosure, and is not intended to limit the disclosure.EXAMPLE 1. Generation and characterization of anti-NCAMl antibodies

[0384] This Example describes the process for generating and characterizing anti-NCAMl antibodies as described herein.

[0385] Upon analyzing the in silico developability of Lorvotuzumab, it was determined that Lorvotuzumab has an abnormally high score for patches of positive charge (PPC), which could contribute to suboptimal pharmacokinetic properties. As one approach to address this potential liability, sequences of anti-NCAMl antibodies were generated from the anti- NCAMl antibody N901 by grafting the CDRs of N901 onto alternate human frameworks while minimizing positive charge within the framework regions. For the first version, N901 CDRs were grafted to the human framework with highest identity to N901. For the second version, N901 framework residues were mutated to residues that are present at high frequency in human variable genes.

[0386] The variable heavy (VH) and variable light (VL) chain sequences of N901, Lorvotuzumab, and the generated humanized antibodies were analyzed for humanness using the BioPhi algorithm (Table El). Outputted parameters include: (i) OASis percentile, which reports on the humanness of a sequence relative to existing therapeutic antibodies; (ii) OASis identity, which reports on the fraction of 9-mer peptides in the sequence that are considered human; and (iii) the number of non-human peptides within the sequence. Results revealed that the engineered VH and VL domains (vl and v2) had OASis percentile and OASis identity scores higher than those of N901 (mN901), and on par with those of Lorvotuzumab (hN901-v0). Additionally, the number of non-human peptides within the engineered VH and VL sequences was reduced compared to N901.152MF-363628402Attorney Docket No. 22419-20010.40Table El

[0387] As another approach to reduce positive charge within Lorvotuzumab variable domains, mutations were made within CDRs to remove positively charged arginine or lysine residues. Mutations include R96Q and K97Q within CDR3 of the VH, and K50Q within CDR2 of the VL, positions numbered according to Kabat. The h- prefix indicates mutations in the VH, and the k- prefix indicates mutations in the VL.

[0388] In silico properties were calculated for all engineered and mutated versions of N901 (Table E2). This analysis revealed that variable domain engineering successfully decreased the isoelectric point (pl) and PPC compared to Lorvotuzumab. It was also discovered that the high PPC score of Lorvotuzumab is primarily caused by the VL domain. All engineered antibodies (Lorvotuzumab, Antibody 7, Antibody 8) that used the original Lorvotuzumab light chain showed high PPC scores, while those antibodies (Antibody 5, Antibody 6, Antibody 1, Antibody 3, Antibody 4, Antibody 2) that used a resurfaced light chain showed a decrease in PPC, restoring this parameter to the value seen for mN901. Point mutation of positively charged residues within CDRs also significantly reduced PPC.Table E2153MF-363628402Attorney Docket No. 22419-20010.40

[0389] Anti-NCAMl antibodies were generated in a CHO expression system. Transiently transfected cell supernatants were harvested on day 7 via centrifugation and filtration. Expression titer was measured by bio-layer interferometry (BLI) using proA sensors. Antibodies were purified on MabSelect Sure resin using typical protein A wash and elution conditions. Purified antibodies were further polished by size-exclusion chromatography (SEC) over Superdex 200 resin at <2g scale, and by cation exchange (CEX) over Poros XS resin at >2g scale. Protein quality and mass accuracy were ensured by analytical SEC-HPLC and mass spectrometry (MS), where final purity was >98% by analytical SEC-HPLC.

[0390] A set of anti-NCAMl antibodies with IgGlk-LALAPG (L234A / L235A / P329G) backbone for reduced binding to Fc gamma receptors and complement proteins was produced for characterization (Table E3). All antibodies were efficiently expressed with transient expression titers ranging from 788.81 mg / L to 2544.91 mg / L. After first step protein A purification, monomeric purity ranged from 93.2% to 98.22%, demonstrating that the antibodies are stable and have low propensity to aggregate. Final % monomer was >99% for all antibodies, which demonstrates that they can be produced with high purity. Certain variants had an increase in titer and / or purity compared to Lorvotuzumab.Table E3154MF-363628402Attorney Docket No. 22419-20010.40

[0391] Another set of antibodies was produced at larger scale with hlgGlk-LALAPA (L234A / L235A / P329A) backbone. Titer and purity results (Table E4) demonstrate that the antibodies can be produced in different IgG backbones without issue.Table E4EXAMPLE 2. Analysis of anti-NCAMl antibody liability

[0392] This Example describes the purification and liability analysis of anti-NCAMl antibodies described herein.

[0393] Liability risk was assessed based on patches of positive charge (PPC) and potential oxidation, deamidation, and isomerization post-translational modification (PTM) sites. In silico analysis revealed several possible PTM motifs, including a DG isomerization motif at D28 and G29 of the VL, an NT deamidation motif at N30 and T31 of the VL, and potential methionine oxidation sites at M95 and Ml 00a of the VH, positions numbered according to Kabat. Based on these risks, a panel of liability mutants was generated as in Example 1, with hlgGlk-LALAPG backbone. Results of the transient expression are summarized in Table E5, and indicate that all antibodies could be expressed efficiently and could be purified to high monomeric purity. The h- prefix indicates mutations in the VH, and the k- prefix indicates mutations in the VL.Table E5155MF-363628402Attorney Docket No. 22419-20010.40EXAMPLE 3: Binding affinity of anti-NCAMl antibodies

[0394] The kinetic association constant (ka), kinetic dissociation constant (kdis), and binding affinity (KD) for each antibody were determined using bio-layer interferometry (BLI) (Table E6) on an Octet RED 384 instrument. All BLI steps were performed in assay buffer containing phosphate-buffered saline (PBS) pH 7.4 with 1% BSA and 0.05% Tween 20. Antibodies were loaded to AHC sensors for 150 seconds at 10 pg / mL followed by a 30- second baseline step. A single 200-nM concentration of antigen protein was associated for 180 seconds followed by a 360-second dissociation step. Regeneration was achieved with five rounds of 10 mM glycine pH 1.5. Fitting was performed on the Octet Analysis software using global fitting to a 1:1 binding model. Purified His-tagged cynomolgus NCAM1 extracellular domain was produced in-house, and was used due to N901’s cross -reactivity with human and cynomolgus NCAM1.

[0395] The results of the BLI assay demonstrate that most variants of N901 showed strong binding to NCAML Certain antibodies had enhanced affinity to NCAM1 compared to Lorvotuzumab. Thus, engineering the framework regions to reduce PPC could improve surface protein properties of hN901 while not disrupting the affinity of the antibody for its antigen. In contrast, the other method of mitigating PPCs [point mutations in CDRs used to reduce positive charge (hR96Q, hK97Q, kK50Q)] tended to reduce antibody affinity. Finally, most CDR point mutants with removed liability motifs had retention of binding affinity, demonstrating that sequence liabilities could be removed without disrupting binding.Table E6156MF-363628402Attorney Docket No. 22419-20010.40EXAMPLE 4. Stress test characterization of anti-NCAMl antibodies

[0396] This Example describes the stress test characteristics of NCAM1 antibodies described herein.

[0397] Lorvotuzumab was tested in a BLI screen to determine stress response at pH 3.0 and pH 8.5. The lower binding response at acidic pH 3.0 indicated potential Asp isomerization at kD28. kD28S and kD28E variants, numbering according to Kabat were generated to address the isomerization liability. Anti-NCAMl antibodies (Antibodies 1, 2, and 3) were stressed at low (pH 3.0) pH at 40 °C for 72 hours. Binding affinity (KD) and % binding activity were determined by BLI (Table E7) based on retention of binding response compared to the unstressed control. Both Lorvotuzumab and its kD28S and kD28E variants had binding157MF-363628402Attorney Docket No. 22419-20010.40 activity that was significantly impacted by acidic stress at 40 °C and had similar KDs at all time points. Surprisingly, mutations introduced to resolve PPCs resulted in a complete rescue of binding activity under the same stress conditions.Table E7

[0398] The antibodies in hlgGlk-LALAPG backbone were analyzed by differential scanning calorimetry (DSC) on a MicroCai DSC to directly assess their thermal stability. Antibodies were heated to 100 °C, and the observed peaks in heat report on protein unfolding. The weight-averaged Tm was calculated by multiplying each of the three observed Tms for each antibody by the fraction of total enthalpy contributed by that peak, and summing. Compared to Lorvotuzumab, the resurfaced variants of Antibody 1, Antibody 2, Antibody 3, and Antibody 5 showed a significant improvement in thermal stability (FIG. 1, Table E8).Table E8. Thermal StabilityEXAMPLE 5: Stability and manufacturing of anti-NCAMl antibodies158MF-363628402Attorney Docket No. 22419-20010.40

[0399] To evaluate the stability of antibodies at high concentrations which could be used during manufacturing, antibodies were concentrated to 20 mg / mL (Concentration 1) and 50 mg / mL (Concentration 2) and analyzed by SEC. Lorvotuzumab, Antibody 3, and Antibody 2 showed good purity at these high concentrations (Table E9), suggesting that these antibodies are manufacturable.Table E9EXAMPLE 6. Binding of anti-NCAMl antibodies to NCAM1 expressing cells

[0400] Anti-NCAMl antibody variants of Antibodies 1, 2, 3, and 5 with resurfacing mutations to address the patches of positive charge (PPC) were evaluated for binding to NCI- H69 cells endogenously expressing NCAM1 by flow cytometry. Mutations within the variable domain but outside of the CDRs of Antibodies 1, 2, 3, and 5 did not impact binding potency compared to Lorvotuzumab (FIG. 2A), whereas additional mutations within the CDRs of Antibody 5 (Antibody 5-K50Q, Antibody 5-K97Q, and Antibody 5-R96Q) resulted in significantly diminished binding compared to unmutated Antibody 5 (FIG. 2B).EXAMPLE 7. Pharmacokinetics of anti-NCAMl antibodies

[0401] Anti-NCAMl antibody variants engineered to reduce PPCs as described in Example 2 were tested in mouse pharmacokinetic experiments. Male CD-I mice were given an i.v. bolus injection of 5 mg / kg in a dose volume of 1 mL / kg of either an Isotype control,Lorvotuzumab, or an antibody described herein with an hlgGl-k Fc region with the LALAPG mutation (Antibody 1, 2, 3, or 5). An exemplary antibody with the K97Q CDR point mutation to reduce a positive charge patch was also tested (Antibody 5-hK97Q). Serum samples were collected at 0.5 hours, 1 hour, 6 hours, 24 hours, 48 hours, 96 hours, 168 hours, 240 hours and 336 hours. The parental Lorvotuzumab antibody showed decreased half-life, increased clearance and decreased area-under-the curve (AUC) compared to an Isotype control antibody. Compared to parental Lorvotuzumab, three of the engineered variants showed increased AUC (Antibodies 2, 3, and 5), while others showed equivalent or reduced AUC (Antibody 1) (Table E10). The serum concentration of the surface engineered variant159MF-363628402Attorney Docket No. 22419-20010.40Antibody 3 remained higher over time compared to Lorvotuzumab and was similar to an Isotype control with normal pharmacokinetics (FIG. 3).

[0402] An anti-NCAMl ADC comprising Antibody 3 (an anti-NCAM antibody comprising a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 10, and an IgGl Fc with LALAPA mutations) and a payload-linker comprising Formula (II) was generated, which is referred to herein as “Antibody 3 ADC.” It should be appreciated that Antibody 3 ADC is an exemplary anti-NCAMl ADC, and that other anti-NCAMl ADCs comprising different anti-NCAMl antibodies (such as those provided herein), different linkers (such as those provided herein), and payloads (such as those provided herein), also fall within the scope of the present disclosure. The structure of the payload-linker portion of Antibody 3 ADC is provided below, wherein the payload- linker is conjugated to the antibody via a cysteine of the anti-NCAMl antibody:Table E10.EXAMPLE 8. Topoisomerase I inhibitor payloads

[0403] This Example describes the use of different payloads.

[0404] In vitro hepatocyte clearance, efflux ratio, permeability, and cytotoxicity were measured for different payloads (deruxtecan (DXd), SN-38, belotecan, topotecan, exatecan, YL0014, tirumotecan, rezetecan, and A- 1743332) including a payload with Formula (II)160MF-363628402Attorney Docket No. 22419-20010.40(e.g., payload-linker having the following formula:A payload with Formula (II) had increased cytotoxicity on cancer cells and a distinct combination of hepatocyte clearance, efflux, and permeability properties from other topoisomerase I inhibitors, despite similar biochemical potency as DXd in a cell-free topoisomerase I inhibition assay. Data is shown in Table Ell and FIGs. 7A-7B.Table Ell.4Median IC50 (nM) across 11 cell lines

[0405] ADCs comprising a payload with Formula (II) (the structure of the payload-linker described in Example 7) were also assayed in vitro against ADCs comprising deruxtecan (DXd) to determine bystander activity. A schema of the assay and the resulting data is shown in FIG. 7C. Cells engineered to express either TROP2 antigen or luciferase were co-cultured161MF-363628402Attorney Docket No. 22419-20010.40 at the indicated ratios (0%, 20%, 40%, 60%, or 100% TROP2-expressing) and treated with the indicated ADCs.

[0406] This Example shows that ADCs comprising payloads with Formula (II) exhibit high membrane permeability, which leads to greater cytotoxicity, improved bystander activity, and improved efficacy in tumors with heterogeneous target expression.EXAMPLE 9. In vivo pharmacology and toxicology of anti-NCAMl ADCs

[0407] NCAM1 is highly expressed in certain neuroendocrine tumor types e.g., small cell lung cancer (SCLC), neuroblastoma (NBL), and Merkel cell carcinoma (MCC) (FIGs. 4A and FIG. 4B). Anti-NCAMl ADCs comprising antibody variants engineered to reduce PPCs as described in Example 2 (including Antibody 3) conjugated to a payload-linker comprising Formula (II) as described in Example 8 (including “Antibody 3 ADC”) were tested for in vivo anti-tumor activity in NCI-H526 (ATCC catalog no. CRL-5811) small cell lung cancer cell line-derived mouse xenografts (FIGs. 5A-5B). BALB / c nude mice were inoculated with the NCI-H526 cells. When mean tumor size reached 197.6 mm3, the mice were given a single i.p. dose of vehicle control or an anti-NCAMl ADC (including Antibody 3 ADC). Tumor volumes were measured for 21 days. Mice treated with vehicle control showed an increase in tumor volume over time and required euthanasia before the end of the study period. Mice treated with the Antibody 3 ADC showed near complete to complete inhibition of tumor growth and absence of significant weight loss (FIG. 5A), while other anti-NCAMl ADC variants comprising the same payload-linker as Antibody 3 ADC either did not improve antibody pharmacokinetics as described in Example 7, Table E10, or showed reduced efficacy (FIG. 5B).

[0408] Antibody 3 ADC showed reduced tumor volume in additional cell line-derived xenograft models including in NCI-H810 (a model for large cell neuroendocrine carcinoma; LCNEC; ATCC catalog no. CRL-5816), NCI-H660 (a model for neuroendocrine prostate cancer; NEPC; ATCC catalog no. CRL-5813), QGP-1 (a model for pancreatic neuroendocrine tumor; PNET; JCRB catalog no. JCRB0183), and SK-N-FI (a model for neuroblastoma; NBL; ATCC catalog no. CRL-2142) as shown in FIG. 5C.

[0409] In a patient-derived xenograft model of Merkel cell carcinoma (MCC), mice treated with vehicle control or an isotype control ADC showed an increase in tumor volume over time, while mice treated with Antibody 3 ADC showed near complete to complete inhibition of tumor growth (FIG. 8),162MF-363628402Attorney Docket No. 22419-20010.40

[0410] Anti-NCAMl ADCs were evaluated in a non-human primate toxicology study in cynomolgus monkeys. A Lorvotuzumab ADC comprising the same payload- linker as Antibody 3 ADC, or Antibody 3 ADC, were administered by intravenous infusion on Days 1 and 22. Both Lorvotuzumab ADC and Antibody 3 ADC treated animals exhibited soft / liquid feces which required veterinary care, with an increased severity and incidence for animals given Lorvotuzumab ADC.

[0411] Antibody 3 ADC was also determined to have improved pharmacokinetic exposure in cynomolgus monkeys as shown in FIG. 6A as compared to the Lorvotuzumab ADC. Treatment with Antibody 3 ADC resulted in a greater reduction in tumor volume in the NCL H526 small cell lung cancer (SCLC) model as shown in FIG. 6B as compared to Lorvotuzumab ADC or a conjugate comprising Lorvotuzumab and mertansine (“Lorvotuzumab mertansine”). Overall, Antibody 3 ADC showed a greater reduction in tumor volume with less toxicity compared to Lorvotuzumab ADC.

[0412] While the disclosure has been described with reference to the specific examples and illustrations, changes can be made and equivalents can be substituted to adapt to a particular context or intended use as a matter of routine development and optimization and within the purview of one of ordinary skill in the art, thereby achieving benefits of the disclosure without departing from the scope of what is claimed and their equivalents.163MF-363628402

Claims

Attorney Docket No. 22419-20010.40CLAIMS1. An antibody or antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 9 or at least about 90% sequence identity to SEQ ID NO: 10.

2. An antibody or antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising an amino acid sequence according to SEQ ID NO: 11; and b) a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 9 or at least about 90% sequence identity to SEQ ID NO: 10.

3. An antibody or antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 7 or at least about 99% sequence identity SEQ ID NO: 8; and b) a light chain variable region (VL) comprising an amino acid sequence according to SEQ ID NO: 12.

4. An antibody or antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising at least about 96% sequence identity to SEQ ID NO: 7, at least about 99% sequence identity SEQ ID NO: 8, or an amino acid sequence according to SEQ ID NO: 11; and b) a light chain variable region (VL) comprising an amino acid sequence having at least about 90% sequence identity to the amino acid sequence of SEQ ID NO: 12, and wherein:(i) the VL comprises a substitution at amino acid position 79 (sequential) relative to the reference sequence of SEQ ID NO: 12;164MF-363628402Attorney Docket No. 22419-20010.40(ii) the VL comprises a substitution at amino acid position 82 (sequential) relative to the reference sequence of SEQ ID NO: 12;(iii) the VL comprises substitutions at amino acid positions 79 and 82 (sequential) relative to the reference sequence of SEQ ID NO: 12;(iv) the VL comprises a substitution at amino acid position 74 (Kabat) relative to the reference sequence of SEQ ID NO: 12;(v) the VL comprises a substitution at amino acid position 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12; or(vi) the VL comprises substitutions at amino acid positions 74 and 77 (Kabat) relative to the reference sequence of SEQ ID NO: 12.

5. An antibody or antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and b) a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); wherein the antibody or antigen-binding fragment is humanized.

6. The antibody or antigen-binding fragment thereof of claim 5, comprising: a) a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 9; b) a VH comprising an amino acid sequence according to SEQ ID NO: 8, and a VL comprising an amino acid sequence according to SEQ ID NO: 10; c) a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 10; d) a VH comprising an amino acid sequence according to SEQ ID NO: 8, and a VL comprising an amino acid sequence according to SEQ ID NO: 9;165MF-363628402Attorney Docket No. 22419-20010.40 e) a VH comprising an amino acid sequence according to SEQ ID NO: 11, and a VL comprising an amino acid sequence according to SEQ ID NO: 9; f) a VH comprising an amino acid sequence according to SEQ ID NO: 11, and a VL comprising an amino acid sequence according to SEQ ID NO: 10; g) a VH comprising an amino acid sequence according to SEQ ID NO: 7, and a VL comprising an amino acid sequence according to SEQ ID NO: 12; or h) a VH comprising an amino acid sequence according to SEQ ID NO: 8, and a VL comprising an amino acid sequence according to SEQ ID NO: 12.

7. An antibody or antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and b) a light chain variable region (VL) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6.

8. The antibody or antigen-binding fragment thereof of any one of claims 1-7, wherein the antibody or the antigen-binding fragment thereof is humanized.

9. An immunoconjugate, comprising: a) an antibody or antigen-binding fragment thereof that binds to neural cell adhesion molecule 1 (NCAM1), wherein the antibody or the antigen-binding fragment thereof comprises:(I) i) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1 (SFGMH); (ii) a CDR2 having an amino acid sequence according to166MF-363628402Attorney Docket No. 22419-20010.40SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and ii) a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE);(ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); or(II) i) a VH comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and ii) a VL comprising (i) a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6; and b) a topoisomerase I inhibitor conjugated to the antibody or the antigen-binding fragment thereof using a linker.

10. The immunoconjugate of claim 9, wherein the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:167MF-363628402Attorney Docket No. 22419-20010.40R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and - [(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3;R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-C6haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci- Ce haloalkynyl)-X2;X1is — O— , — S(O)W— , — NH— , — O— (C=O)— , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ;X2is —OR9, —SR9, or — NHR9;R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually hydrogen or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6;R7is hydrogen, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker;168MF-363628402Attorney Docket No. 22419-20010.40R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)pO(CY2)q]tCY2 — X3;R9is hydrogen, — COR8, — CO2R8, — (CO) — NHR8, or the point of attachment to the linker, with the proviso that exactly one of R7and R9is the point of attachment to the linker; and each X3is individually — H, — OH, — SH, or — NH2.

11. The immunoconjugate of claim 10, wherein the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, -OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O-(Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or an unsubstituted Ci-Ce alkyl;R4is a substituted or an unsubstituted — (Ci-Ce alkyl)-X2;X1is — O— ;X2is —OR9;R5and R6are each individually hydrogen; m is 1;169MF-363628402Attorney Docket No. 22419-20010.40 n4is 1 or 2; n5is 0;R7is hydrogen; andR9is the point of attachment to the linker.

12. The immunoconjugate of claim 10 or 11, wherein n4is 1.

13. The immunoconjugate of claim 10 or 11, wherein n4is 2.

14. The immunoconjugate of claim 11, wherein the topoisomerase I inhibitor is represented by Formula (II) having the structure:wherein:R1is methyl;R2is fluoro;R3is hydrogen;R4is a substituted or an unsubstituted -(C2 alkyl)-X2; and n4is 2.

15. The immunoconjugate of claim 11, wherein the topoisomerase I inhibitor is represented by:170MF-363628402Attorney Docket No. 22419-20010.40wherein R9is the point of attachment to the linker.

16. The immunoconjugate of claim 11, wherein the topoisomerase I inhibitor is represented by:wherein R9is the point of attachment to the linker.

17. The immunoconjugate of claim 11, wherein the topoisomerase I inhibitor is represented by:wherein R9is the point of attachment to the linker.171MF-363628402Attorney Docket No. 22419-20010.4018. The immunoconjugate of claim 9, wherein the topoisomerase I inhibitor and the linker, taken together, are represented by:

19. The immunoconjugate of claim 9, wherein the topoisomerase I inhibitor and the linker, taken together, are represented by:172MF-363628402Attorney Docket No. 22419-20010.4020. The immunoconjugate of claim 9, wherein the topoisomerase I inhibitor and the linker, taken together, are represented by:

21. The immunoconjugate of claim 9, wherein the topoisomerase I inhibitor and the linker, taken together, are represented by:173MF-363628402Attorney Docket No. 22419-20010.4022. The immunoconjugate of claim 9, wherein the topoisomerase I inhibitor and the linker, taken together, are represented by:

23. An immunoconjugate having Formula (I), Djnwherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising:(I) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to SEQ ID NO: 1174MF-363628402Attorney Docket No. 22419-20010.40(SFGMH); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2 (YISSGSFTIYYADSVKG); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 3 (MRKGYAMDY); and a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 4 (RSSQIIIHSDGNTYLE); (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 (KVSNRFS); and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6 (FQGSHVPHT); or (II) a VH comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and a VL comprising (i) a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L1isZ1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl;L3is -(CH2)n1-C(=O)- or -(CH2CH2O)n1-(CH2)n1C(=O)-; each n1is individually an integer from 0 to 10;L4is a tetrapeptide residue;L5is absent or -[NH(CH2)n2]n3-;175MF-363628402Attorney Docket No. 22419-20010.40 n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent,D is a drug moiety; and n is an integer from 1 to 10.

24. The immunoconjugate of claim 23, wherein the drug moiety is represented by Formula (II) having the structure:wherein:176MF-363628402Attorney Docket No. 22419-20010.40R1and R2are each individually selected from the group consisting of hydrogen, halogen, -CN, -OR5, -NR5R6, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or unsubstituted Ci-Ce haloalkyl, a substituted or unsubstituted -O-(Ci-Ce alkyl), a substituted or unsubstituted -O-(Ci-Ce haloalkyl), and - [(CY2)pO(CY2)q]tCY3, or a substituted or unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY3;R4is hydrogen, a substituted or an unsubstituted — (Ci-Ce alkyl)-X2, a substituted or an unsubstituted — (Ci-Ce haloalkyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkenyl)-X2, a substituted or an unsubstituted — (Ci-C6haloalkenyl)-X2, a substituted or an unsubstituted — (Ci-Ce alkynyl)-X2, or a substituted or an unsubstituted — (Ci- Ce haloalkynyl)-X2;X1is — O— , — S(O)W— , — NH— , — O— (C=O)— , — NH— (C=O)— , — NH— (C=O)— O— , — NH— (C=O)— NH— , or — NH— S(O)W— ;X2is —OR9, —SR9, or — NHR9;R5and R6are each individually hydrogen, halogen, a substituted or an unsubstituted Ci-Ce alkyl, a substituted or an unsubstituted Ci-Ce haloalkyl, or — [(CY2)PO(CY2)q]tCY.3; m is 1 or 2; n4and n5are each individually 0, 1 or 2, with the proviso that n4and n5are not both 0; each w is 0, 1 or 2; each Y is individually hydrogen or halogen; each p is individually 1, 2, 3, 4, 5, or 6; each q is individually 0, 1, 2, 3, 4, 5, or 6; each t is individually 1, 2, 3, 4, 5, or 6;R7is hydrogen, —COR8, — CO2R8, —(CO)— NHR8, L4, L5, L6, or L7;R8is a substituted or an unsubstituted Ci-Ce alkyl-X3, a substituted or an unsubstituted Ci-Ce haloalkyl-X3, or — [(CY2)PO(CY2)q]tCY2 — X3;177MF-363628402Attorney Docket No. 22419-20010.40R9is hydrogen, — COR8, — CO2R8, — (CO) — NHR8, L4, L5, L6, or L7, with the proviso that exactly one of R7and R9is L4, L5, L6, or L7; and each X3is individually — H, — OH, — SH, or — NH2.

25. The immunoconjugate of claim 24, wherein the drug moiety of Formula (II) is represented by:wherein:R1and R2are each individually selected from the group consisting of hydrogen, halogen, -OR5, an unsubstituted Ci-Ce alkyl, an unsubstituted Ci-Ce haloalkyl and an unsubstituted -O-(Ci-Ce alkyl), or an unsubstituted -O-(CR5R6)m-O- such that R1and R2are taken together form a ring;R3is a hydrogen or an unsubstituted Ci-Ce alkyl;R4is a substituted or an unsubstituted — (Ci-Ce alkyl)-X2;X1is — O— ;X2is —OR9;R5and R6are each individually hydrogen; m is 1; n4is 1 or 2; n5is 0;178MF-363628402Attorney Docket No. 22419-20010.40R7is hydrogen; andR9is L4, L5, L6, or L7;26. The immunoconjugate of claim 24 or 25, wherein n4is 1.

27. The immunoconjugate of claim 24 or 25, wherein n4is 2.

28. The immunoconjugate of claim 24, wherein the drug moiety of Formula (II) is represented by:R1is methyl;R2is fluoro;R3is hydrogen;R4is a substituted or an unsubstituted -(C2 alkyl)-X2; and n4is 2.

29. The immunoconjugate of any one of claims 23-28, wherein:L2is absent; n1is the integer 2 or 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; andL7is absent.179MF-363628402Attorney Docket No. 22419-20010.4030. The immunoconjugate of claim 25, wherein the drug moiety of Formula (II) is represented by:wherein R9is L4, L5, L6, or L7.

31. The immunoconjugate of claim 25, wherein the drug moiety of Formula (II) is represented by:wherein R9is L4, L5, L6, or L7.

32. The immunoconjugate of claim 25, wherein the drug moiety of Formula (II) is represented by:180MF-363628402Attorney Docket No. 22419-20010.4033. The immunoconjugate of claim 23, wherein Formula (I) is:

34. The immunoconjugate of claim 23, wherein Formula (I) is:181MF-363628402Attorney Docket No. 22419-20010.4035. The immunoconjugate of claim 23, wherein Formula (I) is:182MF-363628402Attorney Docket No. 22419-20010.4036. The immunoconjugate of claim 23, wherein Formula (I) is represented by:

37. The immunoconjugate of claim 23, wherein Formula (I) is represented by:183MF-363628402Attorney Docket No. 22419-20010.4038. An immunoconjugate having Formula (I),Ab-[S-L1-L2-L3-L4-L5-L6-L7-D]n(I) wherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising:(I) a heavy chain variable region (VH) comprising (i) a complementarity determining region (CDR) 1 having an amino acid sequence according to (SFGMH) SEQ ID NO: 1;(ii) a CDR2 having an amino acid sequence according to (YISSGSFTIYYADSVKG) SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to (MRKGYAMDY) SEQ ID NO: 3; and a light chain variable region (VL) comprising (i) a CDR1 having an amino acid sequence according to (RSSQIIIHSDGNTYLE) SEQ ID NO: 4; (ii) a CDR2 having an amino acid sequence according to (KVSNRFS) SEQ ID NO: 5; and (iii) a CDR3 having an amino acid sequence according to (FQGSHVPHT) SEQ ID NO: 6; or184MF-363628402Attorney Docket No. 22419-20010.40(II) a VH comprising (i) a CDR1 having an amino acid sequence according to SEQ ID NO: 1; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 2; and (iii) a CDR3 having an amino acid sequence according to any one of SEQ ID NOs: 3, 35, 36, and 43-47; and a VL comprising (i) a CDR1 having an amino acid sequence according to any one of SEQ ID NOs: 4, 38-42, and 48; (ii) a CDR2 having an amino acid sequence according to SEQ ID NO: 5 or 37; and (iii) a CDR3 having an amino acid sequence according to SEQ ID NO: 6;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L1isZ1and Z2are each individually hydrogen, halogen, NO2, -O-(Ci-Ce alkyl), or Ci-Ce alkyl;L3is -(CH2)n1-C(=O)- or -(CH2CH2O)n1-(CH2)n1C(=O)-; each n1is individually an integer from 0 to 10;L4is a tetrapeptide residue;L5is absent or -[NH(CH2)n2]n3-; n2is an integer from 0 to 6; n3is an integer from 0 to 2;L6is absent,185MF-363628402Attorney Docket No. 22419-20010.40wherein D has the structure:and n is an integer from 1 to 10.

39. The immunoconjugate of any one of claims 23-38, wherein n is an integer from 4 to 8.

40. The immunoconjugate of any one of claims 9-39, wherein the VH comprises an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 7 or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 8.

41. The immunoconjugate of any one of claims 9-40, wherein the VL comprises an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 9 or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 10.186MF-363628402Attorney Docket No. 22419-20010.4042. The immunoconjugate of any one of claims 9-41, wherein the antibody or the antigenbinding fragment thereof is humanized.

43. The antibody or the antigen-binding fragment thereof of any one of claims 1 and 3-8 or the immunoconjugate of any one of claims 9-42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 or SEQ ID NO: 8.

44. The antibody or the antigen-binding fragment thereof of any one of claims 1, 2, and 4- 8 or the immunoconjugate of any one of claims 9-43, wherein the VL comprises an amino acid sequence according to SEQ ID NO: 9 or SEQ ID NO: 10.

45. The antibody or the antigen-binding fragment thereof of any one of claims 1 and 5-8 or the immunoconjugate of any one of claims 9-44, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

46. The antibody or the antigen-binding fragment thereof of any one of claims 1 and 4-8 or the immunoconjugate of any one of claims 9-44, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

47. The antibody or the antigen-binding fragment thereof of any one of claims 1 and 4-8 or the immunoconjugate of any one of claims 9-44, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

48. The antibody or the antigen-binding fragment thereof of any one of claims 1 and 5-8 or the immunoconjugate of any one of claims 9-44, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

49. The antibody or the antigen-binding fragment thereof of any one of claims 2 and 5-8 or the immunoconjugate of any one of claims 9-39, 41, 42, and 44, wherein the VH187MF-363628402Attorney Docket No. 22419-20010.40 comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 9.

50. The antibody or the antigen-binding fragment thereof of any one of claims 2 and 4-8 or the immunoconjugate of any one of claims 9-39, 41, 42, and 44, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 11 and the VL comprises an amino acid sequence according to SEQ ID NO: 10.

51. The antibody or the antigen-binding fragment thereof of any one of claims 2 and 5-8 or the immunoconjugate of any one of claims 9-39 and 42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 7 and the VL comprises an amino acid sequence according to SEQ ID NO: 12.

52. The antibody or the antigen-binding fragment thereof of any one of claims 3 and 5-8 or the immunoconjugate of any one of claims 9-39 and 42, wherein the VH comprises an amino acid sequence according to SEQ ID NO: 8 and the VL comprises an amino acid sequence according to SEQ ID NO: 12.

53. The antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-52 or the immunoconjugate of any one of claims 9-52, wherein the antibody comprises a Fc domain comprising one or more modifications that reduces binding to an Fc receptor, reduces binding to complement component Clq, or both binding to an Fc receptor and reduces binding to complement component Clq.

54. The antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-53 or the immunoconjugate of any one of claims 9-53, wherein the antibody comprises an IgGl Fc domain comprising the amino acid modifications L234A / L235A (LALA), L234A / L235A / P329A (LALAPA), L234A / L235A / P329S (LALAPS), or L234A / L235A / P329G (LALAPG).

55. The antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-54 or the immunoconjugate of any one of claims 5-54, wherein the antibody or the antigen-binding fragment thereof binds to human NCAM1 with an affinity of 10’7M to IO10M.188MF-363628402Attorney Docket No. 22419-20010.4056. An immunoconjugate having Formula (I), Djnwherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a heavy chain variable region (VH) comprising the amino acid sequence according to SEQ ID NO: 7; and a light chain variable region (VL) comprising the amino acid sequence according to SEQ ID NO: 10;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L2is absent;L3is -(CH2)n1-C(=O)-; n1is the integer 2;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; and L6is absent;L7is absent;D has the structure:189MF-363628402Attorney Docket No. 22419-20010.40and n is 8.

57. An immunoconjugate having Formula (I), Djnwherein:Ab is an antibody or an antigen-binding fragment thereof that binds neural cell adhesion molecule 1 (NCAM1), comprising: a heavy chain variable region (VH) comprising the amino acid sequence according to SEQ ID NO: 7; and a light chain variable region (VL) comprising the amino acid sequence according to SEQ ID NO: 10;S is a sulfur atom of a cysteine of the antibody or the antigen-binding fragment thereof;L2is absent;L3is -(CH2)n1-C(=O)-; n1is the integer 5;L4is gly-gly-phe-gly;L5is -[NH(CH2)n2]n3-; n2is the integer 1; n3is the integer 1; and L6is absent;190MF-363628402Attorney Docket No. 22419-20010.40L7is absent;D has the structure:and n is 8.

58. A polynucleotide encoding the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-55 or polypeptide components of the immunoconjugate of any one of claims 9-42, 56, and 57.

59. An expression vector comprising the polynucleotide of claim 58.

60. A cell comprising the expression vector of claim 59.

61. A cell capable of producing the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-55 or polypeptide components of the immunoconjugate of any one of claims 9-42, 56, and 57.

62. A pharmaceutical composition comprising the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-55 or the immunoconjugate of any one of claims 9-42, 56, and 57, and a pharmaceutically acceptable excipient, salt, carrier, or diluent.

63. The pharmaceutical composition of claim 62 for use in treating cancer in a subject.

64. A method of treating cancer comprising administering a therapeutically effective amount of the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and191MF-363628402Attorney Docket No. 22419-20010.4043-55 or the immunoconjugate of any one of claims 9-42, 56, and 57 to a subject in need thereof.

65. The method of claim 64, further comprising administering an additional anti-cancer therapy selected from the group consisting of surgery, chemotherapy, radiotherapy, and immunotherapy.

66. The method of claim 64 or 65, further comprising administering to the subject an immune-modulator, activated lymphocyte cell, kinase inhibitor, or a chemotherapeutic agent.

67. The method of any one of claims 64-66, wherein the cancer is a solid cancer.

68. The method of any one of claims 64-67, wherein the cancer is a neuroendocrine cancer.

69. The method of any one of claims 64-68, wherein the cancer is multiple myeloma, glioblastoma, sarcoma, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, or small intestinal neuroendocrine tumor.

70. The method of any one of claims 64-69, wherein the cancer is NCAM1 positive.

71. A method of diagnosing or prognosing cancer in a subject, the method comprising determining the expression level of NCAM1 in a biological samples using the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-55 or the immunoconjugate of any one of claims 9-42, 56, and 57.

72. Use of an effective amount of the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-55 or the immunoconjugate of any one of claims 9-42, 56, and 57 for treating cancer.

73. The use of claim 72, wherein the cancer is a solid cancer.

74. The use of claim 72 or 73, wherein the cancer is a neuroendocrine cancer.192MF-363628402Attorney Docket No. 22419-20010.4075. The use of any one of claims 72-74, wherein the cancer is multiple myeloma, glioblastoma, sarcoma, small cell lung cancer, Merkel cell carcinoma, ovarian cancer, large cell neuroendocrine carcinoma, neuroendocrine prostate cancer, pancreatic neuroendocrine tumor, neuroblastoma, or small intestinal neuroendocrine tumor.

76. A kit comprising the antibody or the antigen-binding fragment thereof of any one of claims 1-8 and 43-55 or the immunoconjugate of any one of claims 9-42, 56, and 57 and instructions for use.

77. A composition comprising a plurality of the immunoconjugates of any one of claims 23-42, 56, and 57, wherein the average n of the plurality of the immunoconjugates is a number from 1 to 10.193MF-363628402

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