Binding domains for mc1 associated with cancer

Abstract

The present disclosure relates to binding domains that bind to MUC1 associated with cancer, and binding moieties comprising such binding domains. The disclosure further relates to the use of such binding domains or binding moieties in the treatment of cancer. The disclosure also relates to nucleic acids encoding such binding domains or binding moieties, as well as vectors and cells comprising the nucleic acids.

Description

[0001] This application is a divisional application with a filing date of December 20, 2022, application number 202280084666.2, and invention name “Binding domain for MUC1 associated with cancer”. Technical Field

[0002] The present disclosure relates to the field of binding domains and binding moieties such as antibodies. In particular, it relates to the field of therapeutic antibodies for treating cancer. More particularly, it relates to binding domains that bind to MUC1 associated with cancer, and binding moieties comprising such binding domains. Background Art

[0003] Mucin1 or MUC1 is a transmembrane glycoprotein that is usually expressed mainly in glandular or luminal epithelial cells. MUC1 forms a physical barrier for lubrication and protection and helps signal transduction (Shimizu M, Yamauchi K. J Biochem. 1982; 91 (2): 515-24). The extracellular domain of MUC1 can extend up to 200-500nm from the cell surface and is highly glycosylated. Its sugar chain branches are negatively charged and can oligomerize, thus forming a physical barrier to protect cells (Nath S, Mukherjee P. Trends Mol Med. 2014; 20 (6): 332-42).

[0004] The MUC1 gene is located at chromosome 1:155,185,824-155,192,916 reverse strand (GRCh38:CM000663.2) and has 29 transcripts. MUC1 also has many different aliases, such as: Mucin 1; cell surface associated; transmembrane; PEM; PUM; EMA; H23Ag; Episialin; cancer antigen 15-3; ADMCKD(1); CD227; MCKD(1); MCD; tumor-associated epithelial membrane antigen; cancer-associated mucin; tumor-associated epithelial mucin; tumor-associated mucin. The external IDs of the MUC1 gene are HGNC:7508NCBI; Entrez Gene:4582; Ensembl:ENSG00000185499; OMIM:158340 and UniProtKB:P15941.

[0005] The MUC1 protein consists of a single polypeptide chain that is post-translationally cleaved by autoproteolysis by the sea urchin sperm protein enterokinase and agrin (SEA) domain into two peptide subunits, called MUC1-C and MUC-N, which are linked via stable non-covalent bonds ( Figure 1; Gao T. et al. Biomed Pharmacother. 2020; 132: 110888). MUC1-C consists of a short extracellular region of 58 amino acids, a hydrophobic transmembrane region of 28 amino acids, and an intracellular cytoplasmic region of 69 amino acids. MUC1-N is located outside the cell and contains a sequence repeat of about 20 amino acids, with the number of repeats varying from 20 to 120, called variable number tandem repeats (VNTR) (HGVTSAPDTRPAPGSTAPPA). VNTR is rich in serine and threonine residues, and each repeat contains five potential O-glycosylation sites, which under normal conditions will cause MUC1 to be heavily glycosylated in a normal O-glycosylation pattern (Bose M, Mukherjee P. Vaccines (Basel). 2020; 8(4): 659, Zhou D. et al. Molecules. 2018; 23(6): 1326).

[0006] During malignant transformation, MUC1 becomes overexpressed, loses its polar and apical distribution properties, and is expressed both on the surface of epithelial cells and in the cytoplasm. In normal cells, O-glycosylation continues to form mature, elongated and branched O-glycans, whereas in cancer cells, these processes are disrupted. Abnormal MUC1 is characterized by incomplete glycosylation of carbohydrate side chains, resulting in the formation of new carbohydrate side chains, including truncated O-glycans, termed Tn antigen (GalNAcα-O-serine / threonine), and its sialylated form, sialyl-Tn antigen (STn antigen), which contains sialic acid α-2,6 linked to GalNAcα-O-serine / threonine ( Figure 1 ; Gao T. et al. Biomed Pharmacother. 2020).

[0007] The expression of tumor-associated carbohydrates (TACAs) is associated with immunomodulation and inhibition of anticancer immune responses, and the expression of sialoglycosides in tumor cells is associated with the patient's metastatic phenotype and poor prognosis (Rodrigues Mantuano N. et al. J Immunother Cancer. 2020; 8(2): e001222, Bhatia R. et al. Cancer Metastasis Rev. 2019; 38(1-2): 223-236, Anandkumar A, Devaraj H. Scand J Immunol. 2013; 78(1): 1-7, Baldus SE. et al. Tumour Biol. 1998; 19(6): 445-53). Tn and STn antigens are often expressed at high levels in certain tumor cell histologies, including gastric cancer (Moreira IB. et al. Cells. 2020; 9(2): 264), colorectal cancer (Ju T, et al. Cancer Biomark. 2014; 14(1): 63-81), pancreatic cancer (Thomas D. et al. J Cell Mol Med. 2019; 23(10): 6885-6896) and breast cancer (Cazet A. et al. Breast Cancer Res. 2010; 12(3): 204).

[0008] The development of therapeutic antibodies against Tn / STn antigens has been reported. It has been reported that Gatipotuzumab (formerly known as PankoMAb-GEX) showed specific binding to cancer cells with high affinity, but was not studied further in clinical trials (Danielczyk A. et al. Cancer Immunol Immunother. 2006; 55(11): 1337-47). Its safety has been reported in a Phase I trial (Fiedler W. et al. Eur J Cancer. 2016; 63: 55-63); however, in a Phase IIb trial of advanced ovarian cancer (OC), Gatipotuzumab did not show a beneficial effect when used as a single agent as a maintenance treatment for recurrent OC (Ledermann J. et al. ESMO Open. 2022 Feb; 7(1): 100311. Epub 2021 Dec 15). Antibody 5E5 is a mouse anti-human MUC1-Tn / STn antibody reported to show specificity for cancer cells ( AL. et al. Glycobiology. 2006; 16(2): 96-107). 5E5 was used to generate a chimeric antigen receptor (CAR) targeting the TnMUC antigen. This CART-TnMUC1 construct consists of a mouse anti-human scFv derived from the monoclonal antibody 5E5, which recognizes an epitope containing the Tn glycan of MUC1 and is being evaluated in clinical trials for the treatment of refractory solid tumor-type malignancies.

[0009] Therefore, there remains a need for novel therapeutic interventions that selectively target cancer-associated MUC1. Summary of the invention

[0010] In certain embodiments, the present invention provides a new pharmaceutical agent for treating human disease, particularly for treating cancer.

[0011] In certain embodiments, the present invention relates to a binding domain that binds to a MUC1 peptide, wherein the MUC1 peptide comprises the amino acid sequence PAPGSTAPPAHGVTSAPDTRPAPG as shown in SEQ ID NO:249, and wherein the threonine (T) residue at position 14 of the MUC1 peptide comprises α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

[0012] In certain embodiments, the present invention relates to a binding domain that specifically binds to MUC-1 associated with cancer, wherein the binding domain binds to an epitope comprising a VTSA motif of the N-terminal domain of MUC1, and wherein the threonine (T) residue is α-GalNAc glycosylated or Neu5Ac-α(2-6)-GalNAc glycosylated.

[0013] In certain embodiments, the invention relates to a binding domain having binding specificity for MUCl associated with cancer, wherein the binding domain comprises a heavy chain variable region as further defined herein.

[0014] In certain embodiments, the invention relates to binding moieties comprising two binding domains as described herein.

[0015] In certain embodiments, the invention relates to a pharmaceutical composition comprising an effective amount of a binding domain or binding portion as described herein.

[0016] In certain embodiments, the invention relates to a binding domain, binding moiety or pharmaceutical composition as described herein for use in therapy.

[0017] In certain embodiments, the invention relates to a binding domain, binding moiety or pharmaceutical composition as described herein for use in treating cancer.

[0018] In certain embodiments, the invention relates to a method of treating a disease comprising administering to an individual in need thereof an effective amount of a binding domain, binding moiety or pharmaceutical composition as described herein.

[0019] In certain embodiments, the invention relates to a method of treating cancer comprising administering to an individual in need thereof an effective amount of a binding domain, binding moiety or pharmaceutical composition as described herein.

[0020] In certain embodiments, the invention relates to a nucleic acid sequence encoding a heavy chain variable region as defined herein.

[0021] In certain embodiments, the invention relates to a cell comprising a nucleic acid sequence encoding a heavy chain variable region as defined herein.

[0022] In certain embodiments, the invention relates to cells that produce a binding domain or binding portion as described herein.

[0023] In certain embodiments, the invention relates to a kit comprising a container containing a binding domain or binding moiety as described herein and, optionally, instructions for use. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The following naming conventions are used herein as follows. In the figures, reference bivalent monospecific antibodies are represented in the format of SEQ ID NO: A / B, wherein SEQ ID NO: A refers to the heavy chain of the two binding domains, and SEQ ID NO: B refers to the light chain of the two binding domains. The bivalent monospecific MUC1 IgG of the invention exemplified herein comprises a light chain as shown in SEQ ID NO: 217 and a CH1 region, hinge region, CH2 region and CH3 region as shown in SEQ ID NO: 223, 222, 224 and 225, respectively, and is represented in the format of SEQ ID NO: A, which refers to the heavy chain variable region of the two binding domains.

[0025] Figure 1 - Schematic diagram of the structure of MUC1. A) The structure of normal MUC1; B) The structure of abnormal MUC1 (adapted from Gao T. et al. Biomed Pharmacother. 2020 Figure 1 ). Cancer-associated (CA) MUC1 epitopes include exposed core peptide, MUC1-Tn, and MUC1-sTn.

[0026] Figure 2- FACS data indicate the specificity and relative affinity of MUC1 IgG for MCF7 cells (Panels A; C; E; G; I; K; M and O) compared to MCF10A cells (Panels B; D; F; H; J; L; N and P). MUC1 IgG includes core binder (CB), Tn binder (TnB) and (S)Tn binder (SEQ ID NO). Negative control antibodies are SEQ ID NO: 216 / 217; positive control antibodies are SEQ ID NO: 210 / 211 (AP) and SEQ ID NO: 212 / 213 (OP). X-axis: IgG concentration in μg / ml; Y-axis: median PE signal x10 4 .

[0027] Figure 3 - Affinity for MUCl-Tn was plotted against affinity for MUCl-STn in a MUCl-(S)Tn binder.

[0028] X-axis: affinity of IgG to MUC1-STn measured in ELISA, expressed as AUC normalized by reference antibody 5E5 and reference analogs of isotype controls. Y-axis: depicts affinity of Fab to MUC1-Tn measured in ELISA, expressed as AUC normalized by reference antibody 5E5 and reference analogs of isotype controls. The squares in the circles represent two groups of antibodies based on very similar HCDR1: 1) SEQ ID NO: 1; SEQ ID NO: 29; SEQ ID NO: 45; SEQ ID NO: 57; SEQ ID NO: 78; 2) antibodies of SEQ ID NO: 5; SEQ ID NO: 9; SEQ ID NO: 178.

[0029] Figure 4 - Glycopeptide array data provide EC50 values ​​(ug / ml) for binding of MUCl IgG to MUCl-Tn glycopeptides T1-T12 and S1-S12. No values ​​indicate that the binding was undetectable.

[0030] Figure 5 -FACS data indicate the affinity of MUC1 IgG to T47D huGalNAc cells expressing MUC1-STn (Figures AD) and T47D cells expressing MUC1-Tn (Figures EH). Negative control antibodies are SEQ ID NO: 216 / 217; positive control antibodies are SEQ ID NO: 210 / 211 (AH) and SEQ ID NO: 212 / 213 (D; H). X-axis: IgG concentration in μg / ml; Y-axis: median fluorescence x10 4 .

[0031] Figure 6 - FACS data indicate the affinity of MUC1 IgG to MC38 huMUC1 COSMC KO cells (Panels A; C; D; F; G; I and J) compared to MC38 huMUC1 cells (Panels B; E; H and K). Negative control antibodies are SEQ ID NO: 216 / 217; positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). X-axis: IgG concentration in μg / ml; Y-axis: median fluorescence x10 4 . DETAILED DESCRIPTION

[0032] In certain embodiments, the present invention provides a new pharmaceutical agent for diagnosis and treatment of disease, particularly in humans, and particularly for diagnosis and treatment of cancer. In certain embodiments, the present invention provides a binding domain that specifically binds to MUC1 associated with cancer, rather than MUC1 expressed on non-cancerous cells. Specifically, in certain embodiments, the present invention provides for the first time a class of binding domains that specifically bind to MUC1 associated with cancer, which comprises Tn (α-GalNAc) and / or STn (Neu5Ac-α(2-6)-GalNAc) glycosylation at a certain amino acid residue or certain amino acid residues of the VTSA motif, which is different from the normally glycosylated MUC1.

[0033] In certain embodiments, the present invention provides a binding domain that binds a MUC1 peptide, wherein the MUC1 peptide comprises the amino acid sequence PAPGSTAPPAHGV T SAPDTRPAPG (SEQ ID NO: 249) or consists thereof, and wherein the threonine (T) residue at position 14 of the peptide (indicated herein in bold and underlined) comprises α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

[0034] In certain embodiments, the present invention provides a binding domain that specifically binds to MUC-1 associated with cancer, wherein the binding domain binds to an epitope comprising a VTSA motif of the N-terminal domain of MUC1, and wherein the threonine (T) residue is α-GalNAc glycosylated or Neu5Ac-α(2-6)-GalNAc glycosylated.

[0035] In certain embodiments, a binding domain of the invention binds to a MUC1 peptide, wherein the MUC1 peptide comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*SAPDTRPAPG (SEQ ID NO: 249), and wherein * represents Tn glycosylation. In certain embodiments, a binding domain of the invention binds to a MUC1 peptide, wherein the MUC1 peptide comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*SAPDTRPAPG (SEQ ID NO: 249), and wherein * represents STn glycosylation.

[0036] In certain embodiments, the binding domain of the present invention comprises Tn or STn glycosylation of the serine (S) residue at position 15 of the MUC1 peptide, in addition to Tn or STn glycosylation of the threonine residue at position 14 of the peptide as shown in SEQ ID NO: 249. In certain embodiments, the binding domain of the present invention binds to a MUC1 peptide, wherein the MUC1 peptide comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*S*APDTR PAPG (SEQ ID NO: 249), wherein * indicates Tn glycosylation. In certain embodiments, the binding domain of the present invention binds to a MUC1 peptide, wherein the MUC1 peptide comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*S*APDTRPAPG (SEQ ID NO: 249), wherein * indicates STn glycosylation. In certain embodiments, the binding domain of the present invention binds to a MUC1 peptide, wherein the MUC1 peptide comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*S*APDTRPAPG (SEQ ID NO: 249), wherein * indicates STn glycosylation. ● APDTRPAPG (SEQ ID NO: 249) or consisting thereof, wherein * indicates Tn glycosylation and ● In certain embodiments, the binding domain of the invention binds to a MUC1 peptide, wherein the MUC1 peptide comprises the amino acid sequence PAPGSTAPPAHGVT*S ● APDTRP APG (SEQ ID NO: 249) or consisting thereof, wherein * represents STn glycosylation and ● Indicates Tn glycosylation.

[0037] Tn glycosylation refers to the presence of an α-GalNAc group; STn glycosylation refers to a Neu5Ac-α(2-6)-GalNAc group.

[0038] In certain embodiments, a binding domain of the invention binds to human MUCl.

[0039] "Binding domain" refers to a protein molecule or a portion of a protein molecule that binds to a target molecule, which can be determined by binding assays well known to those of ordinary skill in the art. For example, the binding domain can be a heavy chain variable region, a heavy chain variable region paired or connected to a light chain variable region, such as an scFv or Fab. In certain embodiments, the binding domain of the present invention is a Fab. In certain embodiments, "Fab" refers to a binding domain comprising a heavy chain variable region and a light chain variable region. In certain embodiments, "Fab" refers to a binding domain comprising a heavy chain variable region, a light chain variable region, a CH1, and a CL region.

[0040] In certain embodiments, the binding domain of the invention specifically binds to MUC1 associated with cancer, meaning that the binding domain does not bind to MUC1 peptides that lack abnormal glycosylation patterns associated with cancer at detectable levels, or only binds to them at much higher concentrations, as determined in a glycopeptide array, particularly a glycopeptide array as described herein. Abnormal glycosylation patterns associated with cancer include Tn glycosylation (α-GalNAc) and STn glycosylation (Neu5Ac-α(2-6)-GalNAc). Thus, in certain embodiments of the invention, the binding domain does not or substantially does not bind to a MUC1 peptide comprising or consisting of an amino acid sequence as shown in SEQ ID NO:249 and the amino acid sequence does not comprise α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

[0041] In certain embodiments, the binding domains of the present invention bind to an epitope comprising the VTSA motif of the N-terminal domain of MUC1. In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGS*TAPPAHGVTSAPDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation. In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGS*TAPPAHGVTSAPD TRPAPG (SEQ ID NO: 249), wherein * indicates STn glycosylation. In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGST*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation. In certain embodiments, the binding domains of the present invention bind to a MUC1 peptide wherein the MUC1 peptide comprises or consists of the amino acid sequence PAPGST*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn glycosylation. In certain embodiments, the binding domains of the present invention do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGST*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), wherein * indicates STn glycosylation. In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGSTAPPAH GVTS*APDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation. In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGSTAPPAHGVTS*APDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn glycosylation. In certain embodiments, a binding domain of the invention does not or substantially does not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGSTAPPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation. In certain embodiments, a binding domain of the invention does not or substantially does not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGSTAPPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), wherein * indicates Tn glycosylation.In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGS*T*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation. In certain embodiments, the binding domains of the present invention also bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGST*APPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), wherein * indicates Tn glycosylation. In certain embodiments, the binding domains of the present invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGST*APPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), wherein * indicates STn glycosylation. In certain embodiments, the binding domains of the invention do not or substantially do not bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGS*T*APPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation. In certain embodiments, the binding domains of the invention also bind to a MUC1 peptide comprising or consisting of the amino acid sequence PAPGSTAPPAH GVT*SAPDT*RPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation.

[0042] In certain embodiments, a binding domain is considered to bind to a MUC1 peptide if it binds to the MUC1 peptide as a bivalent monospecific IgG antibody with a binding signal in a glycopeptide array that is at least about 2-fold, or about 2-3-fold, or about 3-fold greater than a negative isotype control. In certain embodiments, a binding domain is considered to bind to a MUC1 peptide if it binds to the MUC1 peptide in the form of a bivalent monospecific IgG antibody with an EC50 (in ug / ml) in a glycopeptide array of at least less than about 200 or 100 or 50 or 10 or 1 or 0.5, or between about 200-0.001 or 100-0.001 or 50-0.001 or 10-0.001 or 1-0.001 or 0.5-0.001 or 200-0.05 or 100-0.05 or 50-0.05 or 10-0.05 or 1-0.05 or 0.5-0.05. In certain embodiments, a binding domain is considered to bind to a MUC1 peptide if it binds to the MUC1 peptide as a bivalent monospecific IgG antibody and its binding signal in a glycopeptide array is at least 2-fold, or 2-3-fold, or 3-fold higher than a negative isotype control. In certain embodiments, a binding domain is considered to bind to a MUC1 peptide if it binds to the MUC1 peptide as a bivalent monospecific IgG antibody and its EC50 (in ug / ml) in a glycopeptide array is at least less than 200 or 100 or 50 or 10 or 1 or 0.5, or between 200-0.001 or 100-0.001 or 50-0.001 or 10-0.001 or 1-0.001 or 0.5-0.001 or 200-0.05 or 100-0.05 or 50-0.05 or 10-0.05 or 1-0.05 or 0.5-0.05. In certain embodiments, a binding domain is considered to bind to a MUC1 peptide if it binds to the MUC1 peptide as a bivalent monospecific IgG antibody and has a binding signal in a glycopeptide array that is at least about 2-fold, or about 2-3-fold, or about 3-fold greater than a negative isotype control, and has an EC50 (in ug / ml) of at least less than about 200 or 100 or 50 or 10 or 1 or 0.5, or between about 200-0.001 or 100-0.001 or 50-0.001 or 10-0.001 or 1-0.001 or 0.5-0.001 or 200-0.05 or 100-0.05 or 50-0.05 or 10-0.05 or 1-0.05 or 0.5-0.05.In certain embodiments, a binding domain is considered to bind to a MUC1 peptide if it binds to the MUC1 peptide in the form of a bivalent monospecific IgG antibody and its binding signal in a glycopeptide array is at least 2-fold, or 2-3-fold, or 3-fold higher than a negative isotype control, and it has an EC50 (in ug / ml) of at least less than 200 or 100 or 50 or 10 or 1 or 0.5, or between 200-0.001 or 100-0.001 or 50-0.001 or 10-0.001 or 1-0.001 or 0.5-0.001 or 200-0.05 or 100-0.05 or 50-0.05 or 10-0.05 or 1-0.05 or 0.5-0.05.

[0043] In certain embodiments, if the binding domain binds to the MUC1 peptide as a bivalent monospecific IgG antibody, the binding signal in the glycopeptide array is less than about 2-fold, or about 2-3-fold, or about 3-fold higher than the negative isotype control, the binding domain is considered not to bind or substantially not to the MUC1 peptide. In certain embodiments, if the binding domain binds to the MUC1 peptide as a bivalent monospecific IgG antibody, the EC50 (in ug / ml) in the glycopeptide array is greater than about 200, the binding domain is considered not to bind or substantially not to the MUC1 peptide. In certain embodiments, if the binding domain binds to the MUC1 peptide as a bivalent monospecific IgG antibody, the binding signal in the glycopeptide array is less than 2-fold, or 2-3-fold, or 3-fold higher than the negative isotype control, the binding domain is considered not to bind or substantially not to the MUC1 peptide. In certain embodiments, if the binding domain binds to the MUC1 peptide as a bivalent monospecific IgG antibody with an EC50 (in ug / ml) greater than 200 in a glycopeptide array, the binding domain is considered not to bind or substantially not to the MUC1 peptide. In certain embodiments, if the binding domain binds to the MUC1 peptide as a bivalent monospecific IgG antibody with a binding signal in a glycopeptide array that is less than about 2-fold, or about 2-3-fold, or about 3-fold greater than a negative isotype control, the binding domain is considered not to bind or substantially not to the MUC1 peptide. In certain embodiments, if the binding domain binds to the MUC1 peptide as a bivalent monospecific IgG antibody with an EC50 (in ug / ml) greater than about 200 in a glycopeptide array, the binding domain is considered not to bind or substantially not to the MUC1 peptide. In certain embodiments, a binding domain is considered to not or substantially not bind to a MUC1 peptide if it binds to a MUC1 peptide as a bivalent monospecific IgG antibody, has a binding signal in a glycopeptide array that is less than about 2-fold, or about 2-3-fold, or about 3-fold greater than a negative isotype control, and has an EC50 (in ug / ml) greater than about 200. In certain embodiments, a binding domain is considered to not or substantially not bind to a MUC1 peptide if it binds to a MUC1 peptide as a bivalent monospecific IgG antibody, has a binding signal in a glycopeptide array that is less than about 2-fold, or about 2-3-fold, or about 3-fold greater than a negative isotype control, and has an EC50 (in ug / ml) greater than 200.

[0044] For the purposes of this disclosure, whether a binding domain binds to a MUC1 peptide is determined by using a glycopeptide array. The binding data for the binding domains provided herein were obtained using the glycopeptide arrays described in Example 5. Thus, in certain embodiments, the binding of a binding domain of the invention to a MUC1 peptide, or interaction with a VTSA epitope, is determined in a glycopeptide array as described in Example 5.

[0045] Briefly, the glycopeptide array described in Example 5 was performed using the O-glycopeptides listed in Table 2. The array was blocked prior to the addition of MUC1 IgG and control antibodies (ranging from 10 μg / ml to 0.6 ng / ml). The array was covered with an attachment film and shaken at room temperature. The array was then washed and the detection antibody was applied to the array. It was covered from light and shaken at room temperature for one hour, followed by a washing step. The array was read using an Innopsys InnoScan 710 microarray scanner and appropriate software.

[0046] The binding domains or binding portions of the present invention bind to cancer-associated MUC1, which means that they do not bind to normally glycosylated MUC1 on non-tumor cells. As used herein, "binding to cancer-associated MUC1" refers to the typical binding ability of the binding domain or binding portion to cancer-associated MUC1 present on tumor cells or on cell lines modified to express cancer-associated MUC1. The binding of the binding domain or binding portion to the antigen can be assessed in a variety of ways. For the purposes of the present invention, determining whether the binding domain or binding portion binds to cancer-associated MUC1 is achieved by incubating the binding domain or binding portion with cells expressing the cancer-associated MUC1 (e.g., MC38 huMUC1 COSMC KO cells, T47D huGalNAc cells stably expressing human MUC1-STn, and / or T47DWT cells expressing moderate levels of MUC-Tn), as described herein. Unbound binding domains or binding moieties are removed, while bound binding domains or binding moieties are detected, for example, by a labeled antibody that binds to the constant domains of the bound binding domains or binding moieties.

[0047] Antigen binding can be expressed in terms of specificity and affinity. Specificity determines which antigen or epitope thereof is specifically bound by the binding domain or binding portion. Affinity is a measure of the strength of binding to a specific antigen or epitope.

[0048] In certain embodiments, the binding domains of the invention bind to human MUCl which is associated with cancer.

[0049] In certain embodiments, a binding domain of the invention comprises a heavy chain variable region comprising:

[0050] a) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively;

[0051] b) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively;

[0052] c) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively;

[0053] d) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16, respectively;

[0054] e) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively;

[0055] f) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, respectively;

[0056] g) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 26, SEQ ID NO: 27 and SEQ ID NO: 28, respectively;

[0057] h) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32, respectively;

[0058] i) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36, respectively;

[0059] j) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40, respectively;

[0060] k) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:42, SEQ ID NO:43 and SEQ ID NO:44, respectively;

[0061] 1) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively;

[0062] m) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 51 and SEQ ID NO: 52, respectively;

[0063] n) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively;

[0064] o) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 58, SEQ ID NO: 60 and SEQ ID NO: 61, respectively;

[0065] p) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 63, SEQ ID NO: 64 and SEQ ID NO: 65, respectively;

[0066] q) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 67, SEQ ID NO: 68 and SEQ ID NO: 69, respectively;

[0067] r) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:73, respectively;

[0068] s) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 75, SEQ ID NO: 76 and SEQ ID NO: 77, respectively;

[0069] t) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:79, SEQ ID NO:80 and SEQ ID NO:81, respectively;

[0070] u) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 83, SEQ ID NO: 84 and SEQ ID NO: 85, respectively;

[0071] v) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:87, SEQ ID NO:88 and SEQ ID NO:89, respectively;

[0072] w) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:91, SEQ ID NO:92 and SEQ ID NO:93, respectively;

[0073] x) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:95, SEQ ID NO:96 and SEQ ID NO:97, respectively;

[0074] y) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:99, SEQ ID NO:100 and SEQ ID NO:101, respectively;

[0075] z) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 103, SEQ ID NO: 104 and SEQ ID NO: 105, respectively;

[0076] aa) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 107, SEQ ID NO: 108 and SEQ ID NO: 109, respectively;

[0077] bb) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 111, SEQ ID NO: 112 and SEQ ID NO: 113, respectively;

[0078] cc) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 115, SEQ ID NO: 116 and SEQ ID NO: 117, respectively;

[0079] dd) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 119, SEQ ID NO: 120 and SEQ ID NO: 121, respectively;

[0080] ee) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 123, SEQ ID NO: 124 and SEQ ID NO: 125, respectively;

[0081] ff) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128 and SEQ ID NO: 129, respectively;

[0082] gg) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 131, SEQ ID NO: 132 and SEQ ID NO: 133, respectively;

[0083] hh) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 135, SEQ ID NO: 136 and SEQ ID NO: 137, respectively;

[0084] ii) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140 and SEQ ID NO: 141, respectively;

[0085] jj) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 143, SEQ ID NO: 144 and SEQ ID NO: 145, respectively;

[0086] kk) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 147, SEQ ID NO: 148 and SEQ ID NO: 149, respectively;

[0087] 11) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 151, SEQ ID NO: 152 and SEQ ID NO: 153, respectively;

[0088] mm) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 155, SEQ ID NO: 156 and SEQ ID NO: 157, respectively;

[0089] nn) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 159, SEQ ID NO: 160 and SEQ ID NO: 161, respectively;

[0090] oo) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 163, SEQ ID NO: 164 and SEQ ID NO: 165, respectively;

[0091] pp) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 167, SEQ ID NO: 168 and SEQ ID NO: 169, respectively;

[0092] qq) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 171, SEQ ID NO: 172 and SEQ ID NO: 173, respectively;

[0093] rr) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 175, SEQ ID NO: 176 and SEQ ID NO: 177, respectively;

[0094] ss) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 179, SEQ ID NO: 180 and SEQ ID NO: 181, respectively;

[0095] tt) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 183, SEQ ID NO: 184 and SEQ ID NO: 185, respectively;

[0096] uu) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188 and SEQ ID NO: 189, respectively;

[0097] vv) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 191, SEQ ID NO: 192 and SEQ ID NO: 193, respectively;

[0098] ww) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 195, SEQ ID NO: 196 and SEQ ID NO: 197, respectively;

[0099] xx) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200 and SEQ ID NO: 201, respectively;

[0100] yy) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 203, SEQ ID NO: 204 and SEQ ID NO: 205, respectively; or

[0101] zz) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 207, SEQ ID NO: 208 and SEQ ID NO: 209, respectively.

[0102] The heavy chain variable region of the MUCl binding domain of the present invention may contain a limited number of, for example, one, two or three, non-conservative amino acid substitutions, or an unlimited number of conservative amino acid substitutions.

[0103] In certain embodiments, the MUCl binding domains of the present invention also include MUCl binding domain mutants, wherein each HCDR may contain up to three, two, or one amino acid mutation, such as a substitution. In certain embodiments, only one or two HCDRs may contain up to three, two, or one amino acid mutation, such as a substitution. In certain embodiments, such mutants do not contain amino acid mutations in HCDR3. In certain embodiments, the amino acid mutations are conservative amino acid substitutions.

[0104] Generally, conservative amino acid substitutions involve amino acid mutations to homologous amino acid residues, which are residues with similar characteristics or properties. Homologous amino acids are known in the art and are a conventional method for performing amino acid substitutions in an antibody binding domain without significantly affecting the binding or function of an antibody, see, for example, Lehninger principles of biochemistry (Nelson DL, Cox MM and Lehninger AL. New York: WH Freeman, 2017) or Biochemistry (Berg JM, Tymoczko JL and Stryer L. New York: WH Freeman, 2007) and other manuals, all of which are incorporated herein. When determining whether an amino acid can be substituted by conservative amino acids, the following factors can generally be evaluated, such as, but not limited to: (a) the structure of the polypeptide main chain in the substitution region, for example, a folded or helical configuration, (b) the charge or hydrophobicity of the molecule at the target site, and / or (c) the volume of the side chain. If a residue can be substituted by another residue with common characteristics (e.g., similar side chains or similar charge or hydrophobicity), then such a residue is a preferred substituent. For example, the following groups can be identified: (1) nonpolar: Ala (A), Gly (G), Val (V), Leu (L), Ile (I), Pro (P), Phe (F), Trp (W), Met (M); (2) uncharged polar: Ser (S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acidic: Asp (D), Glu (E); (4) basic: Lys (K), Arg (R), His (H). Alternatively, the amino acids can be grouped as follows: (1) aromatic: Phe (F), Trp (W), Tyr (Y); (2) non-polar: Leu (L), Val (V), Ile (I), Ala (A), Met (M); (3) aliphatic: Ala (A), Val (V), Leu (L), Ile (I); (4) acidic: Asp (D), Glu (E); (5) basic: His (H), Lys (K), Arg (R); (6) polar: Gln (Q), Asn (N), Ser (S), Thr (T), Tyr (Y).Alternatively, amino acid residues can be grouped according to their common side chain properties: (1) hydrophobic: Met (M), Ala (A), Val (V), Leu (L), Ile (I); (2) neutral hydrophilic: Cys (C), Ser (S), Thr (T), Asn (N), Gln (Q); (3) acidic: Asp (D), Glu (E); (4) basic: His (H), Lys (K), Arg (R); (5) residues affecting chain orientation: Gly (G), Pro (P); and (6) aromatic: Trp (W), Tyr (Y), Phe (F).

[0105] An amino acid residue is preferably substituted with another amino acid residue present in the same group. Thus, conservative amino acid substitutions may involve exchanging a member of one of these classes for another member of the same class. Typically, the mutation will not or substantially will not result in a loss of binding specificity of the binding domain to its intended target.

[0106] Additional types of amino acid mutations include mutations caused by somatic hypermutation and / or affinity maturation. The binding mutants covered by the present invention include somatic hypermutation or affinity matured heavy chain variable regions, which are derived from heavy chain variable regions of the same VH gene segments as the heavy chain variable regions of the sequences described herein, and the mutants have amino acid mutations, including non-conservative and / or conservative amino acid substitutions in one, two or all three HCDRs. Conventional methods for affinity maturation of antibody binding domains are well known in the art, see, for example, Tabasinezhad M. et al. Immunol Lett. 2019; 212: 106-113.

[0107] Whether an amino acid residue in a CDR and / or framework region can be substituted, for example, with a conservative amino acid residue and without or substantially without loss of binding specificity, can be determined by methods well known in the art. Experimental examples include, but are not limited to, alanine scanning (Cunningham BC, Wells JA. Science. 1989; 244(4908): 1081-5) and deep mutation scanning (Araya CL, Fowler DM. Trends Biotechnol. 2011; 29(9): 435-42). Computer algorithms that can predict the effects of amino acid mutations have also been developed, such as those described in Sruthi CK, Prakash M. PLoS One. 2020; 15(1): e0227621, Choi Y. et al. PLoS One. 2012; 7(10): e46688, and Munro D, Singh M. Bioinformatics. 2020; 36(22-23): 5322-9.

[0108] In certain embodiments, the MUCl binding domains of the present invention also include MUCl binding domain mutants that, in addition to the mutations in the HCDRs described above, also contain one or more mutations in the framework regions. In certain embodiments, the MUCl binding domain of the invention comprises a heavy chain variable region having an amino acid sequence as set forth in any one of SEQ ID NO:1; 5; 9; 13; 17; 21; 25; 29; 33; 37; 41; 45; 49; 53; 57; 62; 66; 70; 74; 78; 82; 86; 90; 94; 98; 102; 106; 110; 114; 118; 122; 126; 130; 134; 138; 142; 146; 150; 154; 158; 162; 166; 170; 174; 178; 182; 186; 190; 194; 198; 202; 206, or having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, the MUCl binding domain mutants of the present invention do not contain mutations in the CDR regions but contain one or more mutations in the framework regions. Such mutants have at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity with the sequences disclosed herein and are expected to retain the binding specificity of MUCl. Thus, in certain embodiments, the MUCl binding domains of the present invention comprise:

[0109] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence shown in SEQ ID NO: 1, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence shown in SEQ ID NO: 2; a HCDR2 of the amino acid sequence shown in SEQ ID NO: 3; and a HCDR3 of the amino acid sequence shown in SEQ ID NO: 4;

[0110] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence shown in SEQ ID NO:5, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence shown in SEQ ID NO:6; a HCDR2 of the amino acid sequence shown in SEQ ID NO:7; and a HCDR3 of the amino acid sequence shown in SEQ ID NO:8;

[0111] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 9, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 10; a HCDR2 of the amino acid sequence of SEQ ID NO: 11; and a HCDR3 of the amino acid sequence of SEQ ID NO: 12;

[0112] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 13, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 14; a HCDR2 of the amino acid sequence of SEQ ID NO: 15; and a HCDR3 of the amino acid sequence of SEQ ID NO: 16;

[0113] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 17, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 18; a HCDR2 of the amino acid sequence of SEQ ID NO: 19; and a HCDR3 of the amino acid sequence of SEQ ID NO: 20;

[0114] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 21, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 22; a HCDR2 of the amino acid sequence of SEQ ID NO: 23; and a HCDR3 of the amino acid sequence of SEQ ID NO: 24;

[0115] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 25, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 26; a HCDR2 of the amino acid sequence of SEQ ID NO: 27; and a HCDR3 of the amino acid sequence of SEQ ID NO: 28;

[0116] - a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 29, wherein the heavy chain variable region comprises HCDR1 of the amino acid sequence shown in SEQ ID NO: 30; HCDR2 of the amino acid sequence shown in SEQ ID NO: 31; and HCDR3 of the amino acid sequence shown in SEQ ID NO: 32;

[0117] - a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:33, wherein the heavy chain variable region comprises HCDR1 of the amino acid sequence shown in SEQ ID NO:34; HCDR2 of the amino acid sequence shown in SEQ ID NO:35; and HCDR3 of the amino acid sequence shown in SEQ ID NO:36;

[0118] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 37, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 38; a HCDR2 of the amino acid sequence of SEQ ID NO: 39; and a HCDR3 of the amino acid sequence of SEQ ID NO: 40;

[0119] - a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:41, wherein the heavy chain variable region comprises HCDR1 of the amino acid sequence shown in SEQ ID NO:42; HCDR2 of the amino acid sequence shown in SEQ ID NO:43; and HCDR3 of the amino acid sequence shown in SEQ ID NO:44;

[0120] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:45, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:46; a HCDR2 of the amino acid sequence of SEQ ID NO:47; and a HCDR3 of the amino acid sequence of SEQ ID NO:48;

[0121] - a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:49, wherein the heavy chain variable region comprises HCDR1 of the amino acid sequence shown in SEQ ID NO:50; HCDR2 of the amino acid sequence shown in SEQ ID NO:51; and HCDR3 of the amino acid sequence shown in SEQ ID NO:52;

[0122] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:53, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:54; a HCDR2 of the amino acid sequence of SEQ ID NO:55; and a HCDR3 of the amino acid sequence of SEQ ID NO:56;

[0123] - a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:57, wherein the heavy chain variable region comprises HCDR1 of the amino acid sequence shown in SEQ ID NO:58; HCDR2 of the amino acid sequence shown in SEQ ID NO:60; and HCDR3 of the amino acid sequence shown in SEQ ID NO:61;

[0124] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:62, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:63; a HCDR2 of the amino acid sequence of SEQ ID NO:64; and a HCDR3 of the amino acid sequence of SEQ ID NO:65;

[0125] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:66, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:67; a HCDR2 of the amino acid sequence of SEQ ID NO:68; and a HCDR3 of the amino acid sequence of SEQ ID NO:69;

[0126] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:70, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:71; a HCDR2 of the amino acid sequence of SEQ ID NO:72; and a HCDR3 of the amino acid sequence of SEQ ID NO:73;

[0127] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:74, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:75; a HCDR2 of the amino acid sequence of SEQ ID NO:76; and a HCDR3 of the amino acid sequence of SEQ ID NO:77;

[0128] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:78, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:79; a HCDR2 of the amino acid sequence of SEQ ID NO:80; and a HCDR3 of the amino acid sequence of SEQ ID NO:81;

[0129] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:82, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:83; a HCDR2 of the amino acid sequence of SEQ ID NO:84; and a HCDR3 of the amino acid sequence of SEQ ID NO:85;

[0130] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO:86, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO:87; a HCDR2 of the amino acid sequence of SEQ ID NO:88; and a HCDR3 of the amino acid sequence of SEQ ID NO:89;

[0131] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 90, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 91; a HCDR2 of the amino acid sequence of SEQ ID NO: 92; and a HCDR3 of the amino acid sequence of SEQ ID NO: 93;

[0132] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 94, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 95; a HCDR2 of the amino acid sequence of SEQ ID NO: 96; and a HCDR3 of the amino acid sequence of SEQ ID NO: 97;

[0133] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 98, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 99; a HCDR2 of the amino acid sequence of SEQ ID NO: 100; and a HCDR3 of the amino acid sequence of SEQ ID NO: 101;

[0134] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 102, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 103; a HCDR2 of the amino acid sequence of SEQ ID NO: 104; and a HCDR3 of the amino acid sequence of SEQ ID NO: 105;

[0135] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 106, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 107; a HCDR2 of the amino acid sequence of SEQ ID NO: 108; and a HCDR3 of the amino acid sequence of SEQ ID NO: 109;

[0136] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 110, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 111; a HCDR2 of the amino acid sequence of SEQ ID NO: 112; and a HCDR3 of the amino acid sequence of SEQ ID NO: 113;

[0137] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 114, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 115; a HCDR2 of the amino acid sequence of SEQ ID NO: 116; and a HCDR3 of the amino acid sequence of SEQ ID NO: 117;

[0138] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 118, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 119; a HCDR2 of the amino acid sequence of SEQ ID NO: 120; and a HCDR3 of the amino acid sequence of SEQ ID NO: 121;

[0139] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 122, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 123; a HCDR2 of the amino acid sequence of SEQ ID NO: 124; and a HCDR3 of the amino acid sequence of SEQ ID NO: 125;

[0140] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 126, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 127; a HCDR2 of the amino acid sequence of SEQ ID NO: 128; and a HCDR3 of the amino acid sequence of SEQ ID NO: 129;

[0141] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 130, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 131; a HCDR2 of the amino acid sequence of SEQ ID NO: 132; and a HCDR3 of the amino acid sequence of SEQ ID NO: 133;

[0142] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 134, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 135; a HCDR2 of the amino acid sequence of SEQ ID NO: 136; and a HCDR3 of the amino acid sequence of SEQ ID NO: 137;

[0143] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 138, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 139; a HCDR2 of the amino acid sequence of SEQ ID NO: 140; and a HCDR3 of the amino acid sequence of SEQ ID NO: 141;

[0144] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 142, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 143; a HCDR2 of the amino acid sequence of SEQ ID NO: 144; and a HCDR3 of the amino acid sequence of SEQ ID NO: 145;

[0145] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 146, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 147; a HCDR2 of the amino acid sequence of SEQ ID NO: 148; and a HCDR3 of the amino acid sequence of SEQ ID NO: 149;

[0146] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 150, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 151; a HCDR2 of the amino acid sequence of SEQ ID NO: 152; and a HCDR3 of the amino acid sequence of SEQ ID NO: 153;

[0147] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 154, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 155; a HCDR2 of the amino acid sequence of SEQ ID NO: 156; and a HCDR3 of the amino acid sequence of SEQ ID NO: 157;

[0148] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 158, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 159; a HCDR2 of the amino acid sequence of SEQ ID NO: 160; and a HCDR3 of the amino acid sequence of SEQ ID NO: 161;

[0149] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 162, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 163; a HCDR2 of the amino acid sequence of SEQ ID NO: 164; and a HCDR3 of the amino acid sequence of SEQ ID NO: 165;

[0150] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 166, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 167; a HCDR2 of the amino acid sequence of SEQ ID NO: 168; and a HCDR3 of the amino acid sequence of SEQ ID NO: 169;

[0151] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 170, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 171; a HCDR2 of the amino acid sequence of SEQ ID NO: 172; and a HCDR3 of the amino acid sequence of SEQ ID NO: 173;

[0152] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 174, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 175; a HCDR2 of the amino acid sequence of SEQ ID NO: 176; and a HCDR3 of the amino acid sequence of SEQ ID NO: 177;

[0153] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 178, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 179; a HCDR2 of the amino acid sequence of SEQ ID NO: 180; and a HCDR3 of the amino acid sequence of SEQ ID NO: 181;

[0154] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 182, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 183; a HCDR2 of the amino acid sequence of SEQ ID NO: 184; and a HCDR3 of the amino acid sequence of SEQ ID NO: 185;

[0155] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 186, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 187; a HCDR2 of the amino acid sequence of SEQ ID NO: 188; and a HCDR3 of the amino acid sequence of SEQ ID NO: 189;

[0156] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 190, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 191; a HCDR2 of the amino acid sequence of SEQ ID NO: 192; and a HCDR3 of the amino acid sequence of SEQ ID NO: 193;

[0157] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 194, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 195; a HCDR2 of the amino acid sequence of SEQ ID NO: 196; and a HCDR3 of the amino acid sequence of SEQ ID NO: 197;

[0158] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence of SEQ ID NO: 198, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence of SEQ ID NO: 199; a HCDR2 of the amino acid sequence of SEQ ID NO: 200; and a HCDR3 of the amino acid sequence of SEQ ID NO: 201;

[0159] - a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO: 202, wherein the heavy chain variable region comprises HCDR1 of the amino acid sequence shown in SEQ ID NO: 203; HCDR2 of the amino acid sequence shown in SEQ ID NO: 204; and HCDR3 of the amino acid sequence shown in SEQ ID NO: 205; or

[0160] - a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% to the amino acid sequence shown in SEQ ID NO: 206, wherein the heavy chain variable region comprises a HCDR1 of the amino acid sequence shown in SEQ ID NO: 207; a HCDR2 of the amino acid sequence shown in SEQ ID NO: 208; and a HCDR3 of the amino acid sequence shown in SEQ ID NO: 209.

[0161] The MUC1 binding domains of the present invention have been generated using a common light chain, in particular a common light chain called VK1-39 / JK1. The binding domains of the present invention may comprise any suitable light chain, including but not limited to common light chains known in the art. In certain embodiments, the MUC1 binding domains of the present invention comprise the common light chain VK1-39 / JK1, or a mutant thereof having a limited number of non-conservative amino acid substitutions, such as one, two or three, or an unlimited number of conservative amino acid substitutions.

[0162] In certain embodiments, the MUCl binding domain of the present invention comprises a light chain variable region having an amino acid sequence as set forth in SEQ ID NO: 218 or a mutant thereof. In certain embodiments, the MUCl binding domain of the present invention comprises a light chain variable region having an amino acid sequence as set forth in SEQ ID NO: 218, or having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto.

[0163] In certain embodiments, the MUCl binding domain of the present invention comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), having the amino acid sequences shown in SEQ ID NO: 219, SEQ ID NO: 220 and SEQ ID NO: 221, respectively. In certain embodiments, the light chain variable region of the MUCl binding domain of the present invention also includes mutants thereof, wherein each LCDR may comprise up to three, two or one amino acid substitutions. The amino acid substitutions may be, for example, conservative amino acid substitutions.

[0164] In certain embodiments, the MUC1 binding domains of the present invention also include MUC1 binding domain mutants that, in addition to the mutations in the LCDR described above, also include one or more mutations in the framework region. In certain embodiments, the MUC1 binding domain mutants of the present invention do not include mutations in the LCDR region but include one or more mutations in the framework region. Such mutants have at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity with the sequences disclosed herein. Thus, in certain embodiments, the MUC1 binding domains of the present invention include:

[0165] - a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity to the amino acid sequence shown in SEQ ID NO:218, wherein the light chain variable region comprises the LCDR1 amino acid sequence shown in SEQ ID NO:219; the LCDR2 amino acid sequence shown in SEQ ID NO:220; and the LCDR3 amino acid sequence shown in SEQ ID NO:221.

[0166] The light chain or light chain variable region comprising these LCDRs and / or light chain variable regions may be a light chain referred to in the art as VK1-39 / JK1. This is a common light chain. The term "common light chain" of the present invention refers to a light chain that can be paired with multiple different heavy chains (e.g., heavy chains with different antigen or epitope binding specificities). The term "common light chain" encompasses light chains that are identical or have some amino acid sequence differences but the binding specificity of the full-length antibody is not affected. For example, within the definition of the common light chain used herein, light chains that are not identical but still functionally equivalent can be prepared or found, for example, by using established mutations that introduce conservative amino acid changes, when paired with heavy chains, amino acid changes and similar mutations in regions that are known or shown to not or only partially contribute to binding specificity.

[0167] In addition to the common light chains comprising the above-mentioned LCDRs and / or light chain variable regions, other common light chains known in the art can be used. Examples of such common light chains include, but are not limited to, VK1-39 / JK5, ​​which comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3), the light chain variable region having an amino acid sequence as shown in SEQ ID NO: 229. In certain embodiments, the light chain comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3), the light chain variable region having an amino acid sequence as shown in SEQ ID NO: 229, wherein each LCDR may comprise up to three, two, or one amino acid mutation, such as a substitution. In certain embodiments, the light chain comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 229, or having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, the light chain comprises a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), which have the amino acid sequences shown in SEQ ID NO: 230, SEQ ID NO: 231 and SEQ ID NO: 232, respectively; VK3-15 / JK1 comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), which has the amino acid sequence shown in SEQ ID NO: 234. In certain embodiments, the light chain comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), which has the amino acid sequence shown in SEQ ID NO: 234, wherein each LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the light chain comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 234, or having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, the light chain comprises a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3), having amino acid sequences as shown in SEQ ID NO: 235, SEQ ID NO: 236, and SEQ ID NO: 237, respectively; VK3-20 / JK1 comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3), and the light chain variable region has an amino acid sequence as shown in SEQ ID NO: 239.In certain embodiments, the light chain comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), wherein the light chain variable region has an amino acid sequence as shown in SEQ ID NO: 239, wherein each LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the light chain comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 239, or having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, the light chain comprises a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), which have the amino acid sequences shown in SEQ ID NO: 240, SEQ ID NO: 241 and SEQ ID NO: 242, respectively; VL3-21 / JL3 comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), wherein the light chain variable region has the amino acid sequence shown in SEQ ID NO: 244. In certain embodiments, the light chain comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), wherein each LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the light chain comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 244, or having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, the light chain comprises a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2), and a light chain CDR3 (LCDR3), having amino acid sequences as shown in SEQ ID NO: 245, SEQ ID NO: 246, and SEQ ID NO: 247, respectively.

[0168] VK1-39 is the abbreviation of immunoglobulin variable kappa 1-39 gene. This gene is also called immunoglobulin kappa mutant 1-39; IGKV139; IGKV1-39; IgVκ1-39. The external ID of this gene is HGNC:5740; EntrezGene:28930; Ensembl:ENSG00000242371. The amino acid sequence of VK1-39 is shown in SEQ ID NO:227. This is the sequence of the V region. This V region can be combined with one of the five J regions. Suitable VJ region sequences are represented as VK1-39 / JK1 (SEQ ID NO: 228) and VK1-39 / JK5 (SEQ ID NO: 229); another name is IgVκ1-39*01 / IGJκ1*01 or IgVκ1-39*01 / IGJκ5*01 (named according to the IMGT database on imgt.org). These names are exemplary and cover allelic mutants of the gene fragment.

[0169] VK3-15 is the abbreviation of immunoglobulin variable Kappa 3-15 gene. The gene is also called immunoglobulin Kappa mutant 3-15; IGKV315; IGKV3-15; IgVκ3-15. The external ID of the gene is HGNC:5816; EntrezGene:28913; Ensembl:ENSG00000244437. The amino acid sequence of VK3-15 is shown in SEQ ID NO:233. This is the sequence of the V region. The V region can be combined with one of the five J regions. A suitable VJ region sequence is represented by VK3-15 / JK1 (SEQ ID NO:234); another name is Vκ3-15*01 / IGJκ1*01 (named according to the IMGT database on imgt.org). These names are exemplary and cover allelic mutants of the gene fragment.

[0170] VK3-20 is the abbreviation of immunoglobulin variable Kappa 3-20 gene. The gene is also called immunoglobulin Kappa mutant 3-20; IGKV320; IGKV3-20; IgVκ3-20. The external ID of the gene is HGNC:5817; EntrezGene:28912; Ensembl:ENSG00000239951. The amino acid sequence of VK3-20 is shown in SEQ ID NO:238. This is the sequence of the V region. The V region can be combined with one of the five J regions. A suitable VJ region sequence is represented as VK3-20 / JK1 (SEQ ID NO:239); another name is IgVκ3-20*01 / IGJκ1*01 (named according to the IMGT database on imgt.org). These names are exemplary and cover allelic mutants of the gene fragment.

[0171] VL3-21 is the abbreviation of immunoglobulin variable Lambda 3-21 gene. This gene is also called immunoglobulin Kappa mutant 3-21; IGLV321; IGLV3-21; IgVλ3-21. The external ID of this gene is HGNC:5905; EntrezGene:28796; Ensembl:ENSG00000211662.2. The amino acid sequence of VK3-21 is shown in SEQ ID NO:243. This is the sequence of the V region. The V region can be combined with one of the five J regions. A suitable VJ region sequence is represented as VL3-21 / JL3 (SEQID NO:244); another name is IgVλ3-21 / IGJλ3 (named according to the IMGT database on imgt.org). These names are exemplary and cover allelic mutants of the gene fragment.

[0172] Furthermore, any light chain variable region of a MUCl antibody available in the art, or any other light chain variable region readily available, such as by antibody display libraries formed by exhibiting antigen binding activity when paired with a MUCl binding domain of the present invention, may be used.

[0173] In certain embodiments, the MUCl binding domain of the binding moiety of the invention may further comprise a CH1 and CL region. Any CH1 domain may be used, in particular a human CH1 domain. An example of a suitable CH1 domain is provided by the amino acid sequence set forth in SEQ ID NO:223. Any CL domain may be used, in particular a human CL. An example of a suitable CL domain is provided by the amino acid sequence set forth in SEQ ID NO:226.

[0174] In one embodiment, the invention provides a MUCl binding domain comprising:

[0175] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively; and

[0176] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0177] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0178] In one embodiment, the invention provides a MUCl binding domain comprising:

[0179] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively; and

[0180] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0181] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0182] In one embodiment, the invention provides a MUCl binding domain comprising:

[0183] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; and

[0184] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0185] Each HCDR and / or LCDR may contain up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not contain an amino acid mutation.

[0186] In one embodiment, the invention provides a MUCl binding domain comprising:

[0187] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16, respectively; and

[0188] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0189] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0190] In one embodiment, the invention provides a MUCl binding domain comprising:

[0191] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; and

[0192] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0193] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0194] In one embodiment, the invention provides a MUCl binding domain comprising:

[0195] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, respectively; and

[0196] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0197] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0198] In one embodiment, the invention provides a MUCl binding domain comprising:

[0199] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 26, SEQ ID NO: 27 and SEQ ID NO: 28, respectively; and

[0200] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0201] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0202] In one embodiment, the invention provides a MUCl binding domain comprising:

[0203] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32, respectively; and

[0204] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0205] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0206] In one embodiment, the invention provides a MUCl binding domain comprising:

[0207] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36, respectively; and

[0208] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0209] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0210] In one embodiment, the invention provides a MUCl binding domain comprising:

[0211] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40, respectively; and

[0212] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0213] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0214] In one embodiment, the invention provides a MUCl binding domain comprising:

[0215] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:42, SEQ ID NO:43 and SEQ ID NO:44, respectively; and

[0216] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0217] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0218] In one embodiment, the invention provides a MUCl binding domain comprising:

[0219] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48, respectively; and

[0220] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0221] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0222] In one embodiment, the invention provides a MUCl binding domain comprising:

[0223] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 51 and SEQ ID NO: 52, respectively; and

[0224] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0225] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0226] In one embodiment, the invention provides a MUCl binding domain comprising:

[0227] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO: 56, respectively; and

[0228] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0229] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0230] In one embodiment, the invention provides a MUCl binding domain comprising:

[0231] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 58, SEQ ID NO: 60 and SEQ ID NO: 61, respectively; and

[0232] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0233] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0234] In one embodiment, the invention provides a MUCl binding domain comprising:

[0235] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 63, SEQ ID NO: 64 and SEQ ID NO: 65, respectively; and

[0236] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0237] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0238] In one embodiment, the invention provides a MUCl binding domain comprising:

[0239] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 67, SEQ ID NO: 68 and SEQ ID NO: 69, respectively; and

[0240] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0241] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0242] In one embodiment, the invention provides a MUCl binding domain comprising:

[0243] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:73, respectively; and

[0244] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0245] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0246] In one embodiment, the invention provides a MUCl binding domain comprising:

[0247] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 75, SEQ ID NO: 76 and SEQ ID NO: 77, respectively; and

[0248] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0249] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0250] In one embodiment, the invention provides a MUCl binding domain comprising:

[0251] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:79, SEQ ID NO:80 and SEQ ID NO:81, respectively; and

[0252] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0253] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0254] In one embodiment, the invention provides a MUCl binding domain comprising:

[0255] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 83, SEQ ID NO: 84 and SEQ ID NO: 85, respectively; and

[0256] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0257] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0258] In one embodiment, the invention provides a MUCl binding domain comprising:

[0259] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 87, SEQ ID NO: 88 and SEQ ID NO: 89, respectively; and

[0260] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0261] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0262] In one embodiment, the invention provides a MUCl binding domain comprising:

[0263] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:91, SEQ ID NO:92 and SEQ ID NO:93, respectively; and

[0264] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0265] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0266] In one embodiment, the invention provides a MUCl binding domain comprising:

[0267] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:95, SEQ ID NO:96 and SEQ ID NO:97, respectively; and

[0268] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0269] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0270] In one embodiment, the invention provides a MUCl binding domain comprising:

[0271] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:99, SEQ ID NO:100 and SEQ ID NO:101, respectively; and

[0272] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0273] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0274] In one embodiment, the invention provides a MUCl binding domain comprising:

[0275] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 103, SEQ ID NO: 104 and SEQ ID NO: 105, respectively; and

[0276] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0277] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0278] In one embodiment, the invention provides a MUCl binding domain comprising:

[0279] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 107, SEQ ID NO: 108 and SEQ ID NO: 109, respectively; and

[0280] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0281] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0282] In one embodiment, the invention provides a MUCl binding domain comprising:

[0283] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 111, SEQ ID NO: 112 and SEQ ID NO: 113, respectively; and

[0284] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0285] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0286] In one embodiment, the invention provides a MUCl binding domain comprising:

[0287] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 115, SEQ ID NO: 116 and SEQ ID NO: 117, respectively; and

[0288] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0289] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0290] In one embodiment, the invention provides a MUCl binding domain comprising:

[0291] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 119, SEQ ID NO: 120 and SEQ ID NO: 121, respectively; and

[0292] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0293] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0294] In one embodiment, the invention provides a MUCl binding domain comprising:

[0295] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 123, SEQ ID NO: 124 and SEQ ID NO: 125, respectively; and

[0296] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0297] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0298] In one embodiment, the invention provides a MUCl binding domain comprising:

[0299] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128 and SEQ ID NO: 129, respectively; and

[0300] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0301] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0302] In one embodiment, the invention provides a MUCl binding domain comprising:

[0303] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 131, SEQ ID NO: 132 and SEQ ID NO: 133, respectively; and

[0304] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0305] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0306] In one embodiment, the invention provides a MUCl binding domain comprising:

[0307] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 135, SEQ ID NO: 136 and SEQ ID NO: 137, respectively; and

[0308] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0309] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0310] In one embodiment, the invention provides a MUCl binding domain comprising:

[0311] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140 and SEQ ID NO: 141, respectively; and

[0312] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0313] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0314] In one embodiment, the invention provides a MUCl binding domain comprising:

[0315] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 143, SEQ ID NO: 144 and SEQ ID NO: 145, respectively; and

[0316] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0317] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0318] In one embodiment, the invention provides a MUCl binding domain comprising:

[0319] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 147, SEQ ID NO: 148 and SEQ ID NO: 149, respectively; and

[0320] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0321] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0322] In one embodiment, the invention provides a MUCl binding domain comprising:

[0323] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 151, SEQ ID NO: 152 and SEQ ID NO: 153, respectively; and

[0324] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0325] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0326] In one embodiment, the invention provides a MUCl binding domain comprising:

[0327] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 155, SEQ ID NO: 156 and SEQ ID NO: 157, respectively; and

[0328] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0329] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0330] In one embodiment, the invention provides a MUCl binding domain comprising:

[0331] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 159, SEQ ID NO: 160 and SEQ ID NO: 161, respectively; and

[0332] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0333] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0334] In one embodiment, the invention provides a MUCl binding domain comprising:

[0335] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 163, SEQ ID NO: 164 and SEQ ID NO: 165, respectively; and

[0336] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0337] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0338] In one embodiment, the invention provides a MUCl binding domain comprising:

[0339] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 167, SEQ ID NO: 168 and SEQ ID NO: 169, respectively; and

[0340] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0341] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0342] In one embodiment, the invention provides a MUCl binding domain comprising:

[0343] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 171, SEQ ID NO: 172 and SEQ ID NO: 173, respectively; and

[0344] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0345] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0346] In one embodiment, the invention provides a MUCl binding domain comprising:

[0347] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 175, SEQ ID NO: 176 and SEQ ID NO: 177, respectively; and

[0348] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0349] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0350] In one embodiment, the invention provides a MUCl binding domain comprising:

[0351] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 179, SEQ ID NO: 180 and SEQ ID NO: 181, respectively; and

[0352] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0353] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0354] In one embodiment, the invention provides a MUCl binding domain comprising:

[0355] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 183, SEQ ID NO: 184 and SEQ ID NO: 185, respectively; and

[0356] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0357] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0358] In one embodiment, the invention provides a MUCl binding domain comprising:

[0359] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188 and SEQ ID NO: 189, respectively; and

[0360] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0361] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0362] In one embodiment, the invention provides a MUCl binding domain comprising:

[0363] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 191, SEQ ID NO: 192 and SEQ ID NO: 193, respectively; and

[0364] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0365] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0366] In one embodiment, the invention provides a MUCl binding domain comprising:

[0367] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 195, SEQ ID NO: 196 and SEQ ID NO: 197, respectively; and

[0368] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0369] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0370] In one embodiment, the invention provides a MUCl binding domain comprising:

[0371] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200 and SEQ ID NO: 201, respectively; and

[0372] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0373] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0374] In one embodiment, the invention provides a MUCl binding domain comprising:

[0375] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 203, SEQ ID NO: 204 and SEQ ID NO: 205, respectively; and

[0376] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0377] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0378] In one embodiment, the invention provides a MUCl binding domain comprising:

[0379] - heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 207, SEQ ID NO: 208 and SEQ ID NO: 209, respectively; and

[0380] - a light chain CDR1 (LCDR1) having the amino acid sequence shown in SEQ ID NO: 219, a light chain CDR2 (LCDR2) having the amino acid sequence shown in SEQ ID NO: 220, a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221, and

[0381] Each HCDR and / or LCDR may comprise up to three, two or one amino acid mutations, such as substitutions. In certain embodiments, the HCDR and / or LCDR does not comprise an amino acid mutation.

[0382] In one embodiment, the invention provides a MUCl binding domain comprising:

[0383] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 1, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0384] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0385] In one embodiment, the invention provides a MUCl binding domain comprising:

[0386] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 5, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0387] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0388] In one embodiment, the invention provides a MUCl binding domain comprising:

[0389] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 9, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0390] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0391] In one embodiment, the invention provides a MUCl binding domain comprising:

[0392] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 13, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0393] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0394] In one embodiment, the invention provides a MUCl binding domain comprising:

[0395] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 17, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0396] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0397] In one embodiment, the invention provides a MUCl binding domain comprising:

[0398] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 21, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0399] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0400] In one embodiment, the invention provides a MUCl binding domain comprising:

[0401] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 25, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0402] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0403] In one embodiment, the invention provides a MUCl binding domain comprising:

[0404] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 29, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0405] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0406] In one embodiment, the invention provides a MUCl binding domain comprising:

[0407] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 33, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0408] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0409] In one embodiment, the invention provides a MUCl binding domain comprising:

[0410] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 37, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0411] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0412] In one embodiment, the invention provides a MUCl binding domain comprising:

[0413] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 41, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0414] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0415] In one embodiment, the invention provides a MUCl binding domain comprising:

[0416] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 45, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0417] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0418] In one embodiment, the invention provides a MUCl binding domain comprising:

[0419] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 49, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0420] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0421] In one embodiment, the invention provides a MUCl binding domain comprising:

[0422] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 53, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0423] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0424] In one embodiment, the invention provides a MUCl binding domain comprising:

[0425] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 57, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0426] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0427] In one embodiment, the invention provides a MUCl binding domain comprising:

[0428] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 62, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0429] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0430] In one embodiment, the invention provides a MUCl binding domain comprising:

[0431] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 66, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0432] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0433] In one embodiment, the invention provides a MUCl binding domain comprising:

[0434] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 70, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0435] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0436] In one embodiment, the invention provides a MUCl binding domain comprising:

[0437] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 74, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0438] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0439] In one embodiment, the invention provides a MUCl binding domain comprising:

[0440] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 78, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0441] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0442] In one embodiment, the invention provides a MUCl binding domain comprising:

[0443] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 82, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0444] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0445] In one embodiment, the invention provides a MUCl binding domain comprising:

[0446] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 86, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0447] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0448] In one embodiment, the invention provides a MUCl binding domain comprising:

[0449] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 90, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0450] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0451] In one embodiment, the invention provides a MUCl binding domain comprising:

[0452] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 94, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0453] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0454] In one embodiment, the invention provides a MUCl binding domain comprising:

[0455] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 98, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0456] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0457] In one embodiment, the invention provides a MUCl binding domain comprising:

[0458] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 102, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0459] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0460] In one embodiment, the invention provides a MUCl binding domain comprising:

[0461] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 106, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0462] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0463] In one embodiment, the invention provides a MUCl binding domain comprising:

[0464] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 110, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0465] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0466] In one embodiment, the invention provides a MUCl binding domain comprising:

[0467] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 114, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0468] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0469] In one embodiment, the invention provides a MUCl binding domain comprising:

[0470] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 118, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0471] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0472] In one embodiment, the invention provides a MUCl binding domain comprising:

[0473] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 122, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0474] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0475] In one embodiment, the invention provides a MUCl binding domain comprising:

[0476] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 126, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0477] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0478] In one embodiment, the invention provides a MUCl binding domain comprising:

[0479] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 130, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0480] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0481] In one embodiment, the invention provides a MUCl binding domain comprising:

[0482] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 134, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0483] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0484] In one embodiment, the invention provides a MUCl binding domain comprising:

[0485] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 138, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0486] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0487] In one embodiment, the invention provides a MUCl binding domain comprising:

[0488] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 142, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0489] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0490] In one embodiment, the invention provides a MUCl binding domain comprising:

[0491] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 146, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0492] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0493] In one embodiment, the invention provides a MUCl binding domain comprising:

[0494] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 150, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0495] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0496] In one embodiment, the invention provides a MUCl binding domain comprising:

[0497] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 154, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0498] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0499] In one embodiment, the invention provides a MUCl binding domain comprising:

[0500] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 158, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0501] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0502] In one embodiment, the invention provides a MUCl binding domain comprising:

[0503] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 162, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0504] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0505] In one embodiment, the invention provides a MUCl binding domain comprising:

[0506] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 166, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0507] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0508] In one embodiment, the invention provides a MUCl binding domain comprising:

[0509] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 170, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0510] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0511] In one embodiment, the invention provides a MUCl binding domain comprising:

[0512] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 174, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0513] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0514] In one embodiment, the invention provides a MUCl binding domain comprising:

[0515] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 178, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0516] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0517] In one embodiment, the invention provides a MUCl binding domain comprising:

[0518] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 182, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0519] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0520] In one embodiment, the invention provides a MUCl binding domain comprising:

[0521] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 186, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0522] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0523] In one embodiment, the invention provides a MUCl binding domain comprising:

[0524] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 190, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0525] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0526] In one embodiment, the invention provides a MUCl binding domain comprising:

[0527] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 194, or a heavy chain variable region having a sequence identity of at least 80%, or at least 85%, or at least 90%, or at least 95% thereto; and

[0528] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0529] In one embodiment, the invention provides a MUCl binding domain comprising:

[0530] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 198, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0531] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0532] In one embodiment, the invention provides a MUCl binding domain comprising:

[0533] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 202, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0534] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0535] In one embodiment, the invention provides a MUCl binding domain comprising:

[0536] - a heavy chain variable region having an amino acid sequence as shown in SEQ ID NO: 206, or a heavy chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto; and

[0537] - a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each heavy chain variable region and light chain variable region comprises a HCDR and a LCDR, respectively, which do not comprise amino acid mutations. In certain embodiments, each heavy chain variable region and light chain variable region does not comprise amino acid mutations.

[0538] "Percent (%) identity" of nucleic acid or amino acid sequences referred to herein is defined as the percentage of residues in a candidate sequence that are identical to the residues in a selected sequence after the sequences are aligned for optimal comparison purposes. In order to optimize the alignment between the two sequences, gaps may be introduced in either of the two sequences being compared. Such an alignment may be performed over the full length of the sequences being compared. Alternatively, the alignment may be performed over a shorter length, such as about 20, about 50, about 100 or more nucleic acids / bases or amino acids. Sequence identity is the percentage of consistent matches between two sequences in the reported alignment region.

[0539] The comparison of each sequence and the determination of the percentage of sequence identity between the two sequences can be accomplished using a mathematical algorithm. The skilled person will appreciate that there are several different computer programs that can be used to align two sequences and determine the identity between the two sequences (Kruskal JB. SIAM Review. 1983; 25 (2), 201-237). The Needleman and Wunsch algorithms for aligning two sequences can be used to determine the percentage sequence identity between two amino acid sequences or nucleic acid sequences (Needleman SB, Wunsch CD. J Mol Biol. 1970; 48 (3): 443-53). The Needleman-Wunsch algorithm has been implemented in the computer program NEEDLE. For the purposes of the present invention, the NEEDLE program from the EMBOSS suite is used to determine the percentage of identity of amino acid and nucleic acid sequences (version 2.8.0, Rice P. et al. Trends Genet. 2000; 16 (6): 276-7, http: / / emboss.bioinformatics.nl / ). For protein sequences, EBLOSUM62 is used for substitution matrices. For DNA sequences, DNAFULL was used. The parameters used were a gap opening penalty of 10 and a gap extension penalty of 0.5.

[0540] After the above alignment by the program NEEDLE, the percentage of sequence identity between the query sequence and the sequence of the invention is calculated as follows: the number of corresponding positions showing the same amino acid or the same nucleotide in the two sequences in the alignment minus the total number of aligned gaps, divided by the total length of the alignment.

[0541] In certain embodiments, the invention also provides binding moieties comprising a MUC1 binding domain as described herein. In certain embodiments, the binding moieties of the invention comprise two MUC1 binding domains as described herein. In certain embodiments, the binding moieties of the invention consist of two MUC1 binding domains and an Fc region. In certain embodiments, the Fc region comprises a hinge region, a CH2 and a CH3 domain.

[0542] "Binding moiety" refers to a protein molecule comprising a binding domain, and includes, for example, all antibody formats available in the art, such as full-length IgG antibodies, immunoconjugates, diabodies, BiTEs, Fab fragments, scFv, tandem scFv, single domain antibodies (such as VHH and VH), miniantibodies, scFab, scFv-zipper, nanoantibodies, DART molecules, TandAb, Fab-scFv, F(ab)'2, F(ab)'2-scFv2 and intrabody.

[0543] In certain embodiments, the binding moiety of the present invention is a monospecific binding moiety, in particular a monospecific antibody. The monospecific antibody of the present invention is an antibody in any antibody format that comprises one or more binding domains specific for a single target. In certain embodiments, the monospecific binding moiety of the present invention may further comprise an Fc region or a portion thereof. In certain embodiments, the monospecific binding moiety of the present invention is an IgG1 antibody.

[0544] The Fc region mediates the effector functions of antibodies, such as complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). Depending on the application of the therapeutic antibody or Fc fusion protein, it may be desirable to reduce or increase the effector functions.

[0545] In certain embodiments, the binding moiety of the present invention has Fc effector function. In certain embodiments, the binding moiety of the present invention has enhanced Fc effector function. In certain embodiments, the binding moiety of the present invention exhibits antibody-dependent cell-mediated cytotoxicity (ADCC). The binding moiety (e.g., antibody) can be modified to enhance ADCC activity (for review, see Kubota T et al. Cancer Sci. 2009; 100(9): 1566-72). For example, when the antibody itself has low ADCC activity, the ADCC activity of the antibody can be improved by slightly modifying the constant region of the antibody (Junttila TT. et al. Cancer Res. 2010; 70(11): 4481-9). Sometimes changes are also made to improve preservation or production or to remove the C-terminal lysine (Kubota T et al. Cancer Sci. 2009; 100(9): 1566-72). Another method to increase the ADCC activity of antibodies is to interfere with the glycosylation pathway by enzymes, resulting in a reduction of fucose (von orsten HH. et al. Glycobiology. 2010; 20 (12): 1607-18). Alternatively or additionally, various other strategies can be used to achieve ADCC enhancement, including, for example, glycoengineering (Kyowa Hakko / Biowa, GlycArt (Roche) and Eureka Therapeutics) and mutations, which attempt to increase the binding of Fc to the low-affinity activating FcγRIIIa, and / or reduce the binding to the low-affinity inhibitory FcγRIIb. In certain embodiments, the binding moieties of the present invention exhibit enhanced antibody-dependent cell-mediated cytotoxicity (ADCC). In certain embodiments, the binding moieties of the present invention are afucosylated.

[0546] In certain embodiments, the binding portion of the present invention has a higher binding signal than the reference antibody. In certain embodiments, the binding signal is measured using UC1-Tn expressing cells or MUC-1-STn expressing cells in a FACS assay. In certain embodiments, the MUC1-Tn expressing cells are MCF7 or T47D cells, as described herein. In certain embodiments, the MUC1-Tn expressing cells are huMUC1 COSMC KO cells, which can be produced by stably transfecting MC38 cells with human MUC1 and knocking out the C1galt1c1 (COSMC) gene, as described herein. In certain embodiments, the MUC1-STn expressing cells are T47D huGalNAc cells, which can be produced by stably transfecting T47D cells with plasmids encoding ST6GALNAC, as described herein. In certain embodiments, the reference antibody is a bivalent monospecific antibody comprising two heavy chains of the amino acid sequence shown in SEQ ID NO: 210 and two light chains of the amino acid sequence shown in SEQ ID NO: 211.

[0547] In certain embodiments, the invention provides a binding moiety comprising a MUCl binding domain of the invention, wherein the binding moiety is selected from the group consisting of:

[0548] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 1 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0549] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 5 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0550] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:9 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0551] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 13 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0552] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 17 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0553] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 21 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0554] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 25 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0555] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 29 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0556] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 33 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0557] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 37 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0558] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:41 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0559] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:45 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0560] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:49 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0561] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 53 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0562] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:57 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0563] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:62 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0564] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:66 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0565] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 70 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0566] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 74 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0567] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 78 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0568] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 82 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0569] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:86 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0570] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:90 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0571] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO:94 and one light chain variable region having the amino acid sequence shown in SEQ ID NO:218;

[0572] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 98 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0573] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 102 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0574] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 106 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0575] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 110 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0576] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 114 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0577] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 118 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0578] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 122 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0579] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 126 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0580] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 130 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0581] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 134 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0582] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 138 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0583] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 142 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0584] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 146 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0585] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 150 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0586] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 154 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0587] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 158 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0588] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 162 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0589] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 166 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0590] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 170 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0591] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 174 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0592] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 178 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0593] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 182 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0594] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 186 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0595] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 190 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0596] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 194 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0597] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 198 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0598] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 202 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0599] An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 206 and one light chain variable region having the amino acid sequence shown in SEQ ID NO: 218;

[0600] Or a binding portion that competes with the antibody for binding to a MUCl peptide comprising the amino acid sequence PAPGSTAPPAHGVT*SAPDTRPAPG (SEQ ID NO: 249), and / or PAPGSTAPPAH GVT*S*APDTRPAPG (SEQ ID NO: 249), wherein * indicates Tn or STn glycosylation.

[0601] Further provided herein are nucleic acids that can be used to generate the MUC1 binding domains of the invention or binding portions comprising the MUC1 binding domains. In certain embodiments, such nucleic acids comprise a nucleic acid sequence encoding a heavy chain variable region of a MUC1 binding domain as described herein. In certain embodiments, the nucleic acids of the invention may further comprise a nucleic acid sequence encoding a CH1 region and optionally a hinge region, a CH2 region, and / or a CH3 region. In certain embodiments, the nucleic acids of the invention may further comprise a nucleic acid sequence encoding at least a light chain variable region and optionally a CL region. In certain embodiments, the light chain variable region may be a common light chain variable region as described herein.

[0602] Further provided herein are vectors that can be used to produce the MUC1 binding domain of the present invention or a binding portion comprising a MUC1 binding domain. In certain embodiments, this expression vector comprises a nucleic acid sequence encoding a heavy chain variable region of a MUC1 binding domain as described herein. In certain embodiments, the vector of the present invention may further comprise a vector encoding a nucleic acid sequence of a CH1 region and optionally a hinge region, a CH2 region, and / or a CH3 region. In certain embodiments, the vector of the present invention may further comprise a nucleic acid sequence encoding at least a light chain variable region and optionally a CL region. In certain embodiments, the light chain variable region may be a common light chain variable region as described herein.

[0603] In certain embodiments, the present invention also provides a cell comprising a nucleic acid sequence encoding a heavy chain variable region of a MUC1 binding domain as described herein. In certain embodiments, the cell of the present invention may further comprise a nucleic acid sequence encoding a CH1 region and optionally a hinge region, a CH2 region, and / or a CH3 region. In certain embodiments, the cell of the present invention may further comprise a nucleic acid sequence encoding at least a light chain variable region and optionally a CL region. In certain embodiments, the light chain variable region may be a common light chain variable region as described herein.

[0604] In certain embodiments, the present invention also provides cells that produce a MUC1 binding domain, or a binding portion comprising a MUC1 binding domain, as described herein. In certain embodiments, the cell may be a recombinant cell that has been transformed with a vector of the present invention. In certain embodiments, the cell of the present invention comprises a nucleic acid sequence encoding a heavy chain variable region of a MUC1 binding domain as described herein. In certain embodiments, the cell of the present invention further comprises a nucleic acid sequence encoding a CH1 region and optionally a hinge region, a CH2 region, and / or a CH3 region. In certain embodiments, the cell of the present invention further comprises a nucleic acid sequence encoding at least a light chain variable region, particularly a light chain variable region as described herein, and optionally a CL region.

[0605] In certain embodiments, the present invention provides a pharmaceutical composition comprising an effective amount of a MUCl binding domain, or a binding portion comprising a MUCl binding domain as described herein, and optionally a pharmaceutically acceptable carrier.

[0606] In certain embodiments, the invention provides a MUCl binding domain, or a binding portion comprising a MUCl binding domain, as described herein, and a pharmaceutical composition as described herein, for use in therapy.

[0607] In certain embodiments, the invention provides a MUCl binding domain as described herein, or a binding portion comprising a MUCl binding domain, or a pharmaceutical composition as described herein, for use in treating cancer.

[0608] In certain embodiments, the invention provides a method of treating a disease comprising administering to an individual in need thereof an effective amount of a MUCl binding domain as described herein, or a binding portion comprising a MUCl binding domain, or a pharmaceutical composition as described herein.

[0609] In certain embodiments, the invention provides a method of treating cancer comprising administering to an individual in need thereof an effective amount of a MUCl binding domain as described herein, or a binding portion comprising a MUCl binding domain, or a pharmaceutical composition as described herein.

[0610] As used herein, the terms "individual," "subject," and "patient" are used interchangeably and refer to mammals, such as humans, mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, monkeys, cows, horses, pigs, and the like, particularly humans with cancer.

[0611] As used herein, the term "treatment" refers to any type of intervention or treatment of a subject, or the administration of an active agent or combination of active agents to an individual, with the purpose of treating or improving a disease or its symptoms or producing a positive therapeutic response. As used herein, "positive therapeutic response" refers to a treatment that produces a beneficial effect, such as reversing, alleviating, improving, inhibiting or slowing down symptoms, complications, conditions or biochemical indicators associated with the disease, and preventing the onset, progression, development, severity or recurrence of related symptoms, complications, conditions or biochemical indicators, such as improving a disease or condition, such as at least one symptom of cancer. The beneficial effect may be in the form of an improvement relative to a baseline period, including an improvement relative to a measurement or observation made before the method begins treatment. For example, a beneficial effect may be in the form of slowing down, stabilizing, stopping or reversing the progression of cancer in a subject at any clinical stage, such as evidence of a reduction or elimination of clinical or diagnostic symptoms of the disease or cancer markers. Effective treatment may be, for example, reducing tumor size, reducing the presence of circulating tumor cells, reducing or preventing tumor metastasis, slowing down or stopping tumor growth, and / or preventing or delaying tumor recurrence or recurrence.

[0612] The term "therapeutic amount" or "effective amount" refers to the amount of an agent or combination of agents that treats a disease such as cancer. In some embodiments, a therapeutic amount is an amount sufficient to delay tumor development. In some embodiments, a therapeutic amount is an amount sufficient to prevent or delay tumor recurrence.

[0613] As used herein, an effective amount of an agent or composition is an amount that can, for example: (i) reduce the number of cancer cells; (ii) reduce tumor size; (iii) inhibit, delay, slow to some extent and possibly prevent cancer cell infiltration into peripheral organs; (iv) inhibit tumor metastasis; (v) inhibit tumor growth; (vi) prevent or delay the occurrence and / or recurrence of tumors; and / or (vii) alleviate one or more symptoms associated with cancer to some extent.

[0614] The effective amount may vary according to factors such as the disease state, age, sex, and weight of the individual to be treated, and the ability of the agent or combination of agents to elicit a desired response in the individual, which can be readily assessed by an ordinary physician or other medical professional.

[0615] An effective amount can be administered to a subject in one or more administrations.

[0616] An effective amount can also include an amount that balances any toxic or detrimental effects of the agent or combination of agents against the beneficial effects.

[0617] The term "agent" refers to a therapeutically active substance, in this case a binding domain or binding moiety of the invention, or a pharmaceutical composition of the invention.

[0618] As used herein, "comprise," "comprising" and its conjugations are used in their non-limiting sense, meaning that items following the word are included, but items not specifically mentioned are not excluding.

[0619] The articles "a" and "an" are used herein to refer to one or more of the grammatical object of the article. For example, "an element" refers to one or more elements.

[0620] The reference herein to a patent document or other matter shall not be construed as an admission that the document or matter is known or that the information it contains was part of the public knowledge at the priority date of any claim relating to the patent application.

[0621] All patent and literature references cited in this specification are incorporated herein by reference in their entirety

[0622] Please note that in this specification, unless otherwise stated, the amino acid positions assigned to the CDRs and frameworks in the variable regions of antibodies or antibody fragments are designated according to the Kabat numbering (see Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md., 1987 and 1991)). Amino acids in the constant region are represented according to the EU numbering system.

[0623] The accession number is provided mainly to provide further target identification methods. The actual sequence bound to the protein may be different, for example due to mutations in the encoding gene, such as mutations that occur in certain cancers. The antigen binding site of the binding domain or binding portion of the present invention can bind to the antigen and its various mutants, such as those expressed by certain antigen-positive immune cells or tumor cells. HGNC stands for HUGO Gene Nomenclature Committee. The number after the abbreviation is the accession number, which can be used to retrieve information about the gene and the protein encoded by the gene from the HGNC database. Entrez Gene provides accession numbers or gene IDs, which can be used to retrieve information about the gene or the protein encoded by the gene from the NCBI (National Center for Biotechnology Information) database. Ensembl provides accession numbers, which can obtain information about the gene or the protein encoded by the gene from the Ensembl database. Ensembl is a joint project between EMBL-EBI and the Wellcome Trust Sanger Institute, which aims to develop software systems that can generate and maintain automatic annotations on selected eukaryotic genomes.

[0624] When a gene or protein is mentioned herein, it is preferably a human form of the gene or protein. When a gene or protein is mentioned herein, it refers to both the native gene or protein and a mutant form of the gene or protein that can be detected in a tumor, cancer or the like or can be detected in a human tumor, cancer or the like.

[0625] Example

[0626] In an example used to illustrate the present invention but not intended to limit the present invention in any way, the MUC binding domains of the present invention are characterized as IgG forms, wherein each MUC1 binding domain comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, and a light chain constant region having an amino acid sequence as shown in SEQ ID NO: 226, and is in Fab form. In an example used to illustrate the present invention but not intended to limit the present invention in any way, unless otherwise stated, MUC1 IgG is screened out as IgG1 form, which comprises CH1 having an amino acid sequence as shown in SEQ ID NO: 223, CH2 having an amino acid sequence as shown in SEQ ID NO: 224, and CH3 having an amino acid sequence as shown in SEQ ID NO: 225.

[0627] The reference antibodies and control antibodies used in the examples include:

[0628] - Reference antibody 5E5, which is an analog of the bivalent monospecific humanized antibody 5E5, comprising two heavy chains with the amino acid sequence shown in SEQ ID NO: 210, and two light chains with the amino acid sequence shown in SEQ ID NO: 211. Humanized 5E5 binds to the MUC1-(s)Tn(GS*T*A) epitope.

[0629] - Reference antibody pankomab, which is an analog of the bivalent monospecific antibody pankomab, comprising two heavy chains of the amino acid sequence shown in SEQ ID NO: 212, and two light chains of the amino acid sequence shown in SEQ ID NO: 213. Pankomab binds to the MUC1-(s)Tn(PDT*RP) epitope.

[0630] - Reference antibody HT186-D11, which is an analog of the bivalent monospecific antibody HT186-D11, comprising two heavy chains with the amino acid sequence shown in SEQ ID NO: 214, and two light chains with the amino acid sequence shown in SEQ ID NO: 215. HT186-D11 binds to the MUC1 core (RPAP) epitope.

[0631] - A bivalent monospecific control anti-STn antibody strain 3F1 (SBH Sciences, catalog number SBH-ASTn), which binds to STn.

[0632] - A bivalent monospecific anti-Tn antibody strain 5F4 (SBH Sciences, catalog number SBH-Tn), which binds to Tn.

[0633] - A bivalent monospecific control antibody, VU4H5 (BioKE, Cat. No. 4538S), which binds to the MUCl core (APDTRPAP) epitope.

[0634] - a bivalent monospecific isotype control IgG comprising two heavy chains having the amino acid sequence shown in SEQ ID NO: 216, and two light chains having the amino acid sequence shown in SEQ ID NO: 217.

[0635] Example 1 - Generation of MUC1 Fab

[0636] comprising a binding domain and an antibody that binds to the heavy chain variable region of MUC-1 associated with cancer, which is expressed by transgenic mice ( Several immunization methods were used, including the use of human MUC1 conjugated to KHL. 40 Tn8 or MUC1 40 Tn 10 Peptides are added to cells overexpressing human MUC1-(S)Tn for immunization. Phage display libraries are generated and screened in ELISA and FACS using MUC1 peptides and various cell lines to identify Fabs that bind to MUC1-Tn and / or MUC1-STn (MUC1-(S)Tn). Binders are then recloned into IgG format for further characterization. Once identified and sequenced by the techniques provided herein, the binding domain sequences described herein can then be obtained by any method known in the art.

[0637] Example 2 - Specificity and relative affinity of MUCl-(S)Tn measured using ELISA

[0638] The MUC1 IgG panel was screened in an ELISA to characterize their binding specificity and relative affinity to different MUC1 peptides. Negative assay controls included only the mimetic peptide and secondary antibody. Isotype control antibodies and analogs of the reference antibody 5E5 served as negative and positive antibody controls, respectively.

[0639] Biotinylated MUC1 and control peptides were captured onto Maxisorp plates coated with neutravidin (ThermoFisher Scientific, catalog number 31000) at the concentrations shown in Table 1 and allowed to stand at room temperature for one hour. MUC1 IgG and control antibodies were diluted in blocking buffer, added at 10, 1, 0.1, and 0.01 ug / ml, and allowed to stand at room temperature for one hour. Afterwards, the samples were incubated with diluted HRP-conjugated secondary antibodies (Bethyl Laboratories, A80-104P, 1:2000) at room temperature for one hour. Three washing steps were performed using PBST after each standing step. The reaction was terminated using 1M H2SO4, and the signal was developed using TMB peroxidase substrate solution (BD Biosciences, catalog number 555214). The plate was read using a microplate reader at A 450 The following analysis is carried out.

[0640] Table 1. Peptides used in ELISA.

[0641] MUC1 peptide Sequence and (s)Tn modification Modification Capture concentration (ug / ml) MUC140 PAPGSTAPPAHGVTSAPDTRPAPGSTAPPAHGVTSAPDTR none 2 MUC140-Tn10 PAPGS*T*APPAHGVT*S*APDT*R PAPGS*T*APPAHGVT*S*APDT*R Tn 2 MUC140-sTn10 PAPGS*T*APPAHGVT*S*APDT*R PAPGS*T*APPAHGVT*S*APDT*R tT 10 Peptide Mimics Non-MUC1 sequences none 2

[0642] The assay recognizes many antibodies in the MUC1 IgG panel that bind to MUC1 40 Tn 10 The binding specificity of MUC1-(S)Tn antibodies can be divided into three groups: 1) MUC1 core peptide binders (binding to MUC1 40 and MUC1 40 Tn 10 and MUC1 40 STn 10 ); 2) MUC1-Tn binder (binds to MUC1 40 Tn 10 But does not bind to MUC1 40 STn 10 ); and 3) MUC1-(S)Tn binder (binds to MUC1 40 Tn 10 and MUC1 40 STn 10 ).

[0643] Example 3 - Specificity and relative affinity of MUCl-(S)Tn measured using FACS

[0644] The MUC1 IgG panel was used to screen MCF7 cells expressing cancer-associated MUC1-Tn and MCF10A cells expressing wild-type MUC1. Reference antibody pankomab analogs were included as controls for relative expression of MUC1 in different cell lines. The 293FF (MUC1 negative) cell line was included as a negative assay control. Isotype control antibodies and analogs of reference antibody 5E5 were used as negative and positive antibody controls, respectively. Starting from 10ug / ml, the MUC1 IgG panel and control antibodies were titrated using an 8-step 4-fold serial dilution method. Binding to 293FF cells was tested at 10ug / ml.

[0645] MCF7 cells (DSMZ, ACC115), MCF10A cells (ATCC, catalog number CRL-10317) or 293FF cells (Invitrogen, p / n51-0029) were incubated on ice for 30 minutes with MUC1 and control antibodies diluted in FACS buffer (PBS + 0.5% BSA + 2mM EDTA), followed by two washing steps with ice-cold FACS buffer and staining with anti-human IgG-PE (Invitrogen, #H10104) at 1:100. Binding was measured using iQue Screener PLUS. The IntelliCyt The mean fluorescence intensity (MFI) values ​​of each MUC1 antibody were plotted against IgG concentration, and the area under the curve (AUC) was calculated, normalized using isotype antibodies and reference antibody 5E5 analogs as negative and positive antibody controls, respectively.

[0646] The results are as follows Figure 2 As shown. Positive and negative control antibodies showed binding to MCF7, MCF10A and 293FF cells as expected. MUC1 core peptide binders (CB) bound to MCF7 and MCF10A cells, with a few showing low binding to 293FF cells. MUC1-(S)Tn binders (SEQ ID NOs) specifically bound to MCF7, but not to MCF10A cells. MUC1-Tn binders (TnB) specifically bound to MCF7, but not to MCF10A cells; however, had lower affinity compared to MUC1-(S)Tn binders. The affinity of MUC1-(S)Tn binders for MCF7 cells was divergent, with many antibodies showing higher binding signals than the reference antibody 5E5 analog.

[0647] Example 4 - Measurement of the effect of MUC1 Fab panels on MUC1 using ELISA 40 Tn 10 and MUC1 40 STn10 Affinity

[0648] MUC1-(S)Tn IgG plate was cleaved by GingisKHAN to generate Fab (>99% monomer). Biotinylated MUC1 40 Tn 10 and MUC1 40 STn 10 Captured on plates coated with neutravidin at a concentration of 2 μg / ml. A Fab fragment of the reference antibody 5E5 analog was included as a positive control antibody, and a Fab fragment of an isotype control antibody was included as a negative control antibody.

[0649] The MUC1-(S)Tn Fabs plate and control Fabs were diluted 10 times in 8 steps, starting from 10ug / ml, and left to stand at room temperature for one hour, followed by standing at room temperature for one hour with diluted HRP-conjugated secondary antibody (Becton Dickinson, catalog number 555788). Three washing steps were performed using PBST after each standing step. The reaction was terminated using 1M H2SO4, and the signal was developed using TMB peroxidase substrate solution (BD Biosciences, catalog number 555214). The plate was analyzed using a microplate reader at A450.

[0650] MUC1-(S)Tn Fab against MUC1 40 Tn 10 and MUC1 40 STn 10 The affinity of the peptide was similar to that of IgG (data not shown). A general correlation was found between the affinity for the MUC1-Tn peptide and the MUC1-STn peptide (see Figure 3 Many clones showed high affinity for MUC1-Tn and MUC1-STn. In this group, many of the Fabs that showed the highest binding affinity for MUC1-Tn and MUC1-STn shared the same HCDR1 sequence (in Figure 3 (circled in the middle).

[0651] Example 5 - Glycopeptide Array

[0652] The MUC1 IgG panel was screened in the glycopeptide array to determine their binding to the O-glycopeptides listed in Table 2. Background buffer (print buffer) was included as a negative assay control. Positive assay controls were human IgG and mouse IgG. Control antibodies included: reference antibody 5E5 analog, reference antibody VU4H5, 5F4 and 3F1 analogs. MUC1 and control antibodies were tested in 8 steps of 4-fold serial dilutions starting at 10ug / ml.

[0653] O-glycan array assay was performed according to the manual instructions (Z Biotech, LLC). The array was blocked for 30 minutes using glycan array blocking buffer (GAAB, Z Biotech Item#10106). Afterwards, the array was washed 3 times using TBS-T-based glycan array assay buffer (GAAB, Z Biotech Item#10107). MUC1 IgG and control antibodies were diluted in GAAB in a range of 10 μg / ml to 0.6 ng / ml and then directly applied to the array. The array was covered with an attachment membrane and shaken at 80 rpm for 1 hour at room temperature, and then the array was washed again 3 times with GAAB. Detection antibodies, Cy3-conjugated anti-human IgG and Cy3-conjugated anti-mouse IgG were diluted in GAAB and applied to the array. Covered and protected from light and shaken at 80 rpm for 1 hour at room temperature, then washed 3 times with GAAB and washed 2 times with MilliQ water. The array was read at 1 PMT using an Innopsys InnoScan 710 microarray scanner with a high power laser. The software was used to detect each spot on the array and calculate the relative fluorescence unit (RFU) intensity of each spot. The background RFU was subtracted from the RFU value of each spot. The median of each glycan spot was determined and plotted as the reported value.

[0654] Table 2. O-glycopeptides used for glycopeptide array

[0655]

[0656] All control antibodies bound to their described epitopes. Results for MUC1 IgG are shown in Figure 4 As shown. MUC1 IgG previously identified as a MUC1-(S)Tn binder showed strong binding to peptides T4, T9, T11, S4, S9, and S11. These peptides all contain modifications of the threonine residue in the VTSA motif, either Tn modification (α-GalNAc glycosylation) or STn modification (Neu5Ac-α(2-6)-GalNAc glycosylation). With the exception of peptide T8, none of the other peptides contained modifications at this position, and the MUC1-(S)Tn binder did not bind to the peptides. The MUC1-(S)Tn binder also bound to the peptides when the peptides contained, in addition to the (S)Tn modification of the threonine residue in the VTSA motif, Tn or STn modifications of the serine residue in the VTSA motif (T9, S9) or Tn or STn modifications of the threonine residue in the PDTR motif (T11, S11).

[0657] MUC1 IgG previously identified as a MUC1-Tn binder showed strong binding to peptides T2, T7, and T10. These peptides all contain a Tn modification (α-GalNAc glycosylation) of a serine residue in the GSTA motif. None of the other peptides contain a modification at this position, and the MUC1-(S)Tn binder does not bind to the peptide. The MUC1-Tn binder also binds to the peptide when the peptide contains, in addition to the Tn modification of the serine residue in the GSTA motif, a Tn modification of a threonine residue in the GSTA motif (T7) or a Tn modification of a threonine residue in the GSTA motif and a Tn modification of a threonine residue in the PDTR motif (T10).

[0658] Example 6 - Affinity of MUC1 IgG for T47D huGalNAc cells

[0659] MUC1 IgG was screened for binding to T47D huGalNAc cells that stably express human MUC1-STn and T47D WT cells that express intermediate levels of MUC-Tn. An isotype control antibody was included as a negative control antibody, and a reference antibody 5E5 analog was included as a positive control antibody. A reference antibody pankomab analog was included as a control for the relative expression of MUC1 in different cell lines. Other control antibodies included 3F1 and 5F4. MUC1, isotype control, 5E5 analog, and pankomab antibody analog were titrated using 8-step 4-fold serial dilutions starting at 10ug / ml; 3F1 and 5F4 were used at 10ug / ml.

[0660] T47D cells (Sigma-Aldrich, catalog number 85102201-1VL) and T47D huGalNAc cells (generated by stably transfecting T47D cells with plasmids encoding ST6GALNAC1 (accession number Q9NSC7; Uniprot)) were placed on ice for 30 minutes with MUC1 IgG and control antibodies diluted in FACS buffer (PBS + 0.5% BSA + 2mM EDTA), followed by two washes with ice-cold FACS buffer and stained with 1:100 anti-human IgG-PE secondary antibody (Invitrogen, catalog number H10104) (for MUC1, isotype control, 5E5 analog and pankomab antibody sample analog), or 1:400 anti-mouse IgG-PE secondary antibody (Invitrogen, catalog number M30004-4) (for 3F1 and 5F4 antibody samples). Binding was measured using iQue Screener PLUS. Using IntelliCyt The MFI values ​​of each MUC1 antibody were plotted against IgG concentrations, and the area under the curve (AUC) was calculated, normalized using isotype antibodies and reference antibody 5E5 analogs as negative and positive antibody controls, respectively.

[0661] The results are shown in Figure 5 The panel of MUC1(S)Tn binders showed divergent binding affinities to T47D huGalNAc cells. Many antibodies showed higher maximal binding than the analogs of the reference antibody 5E5. The affinity to T47D huGalNAc cells was closely correlated with the affinity to the MUC1-STn peptide.

[0662] The panel of MUC1-(S)Tn binders showed divergent binding affinities to T47D WT cells. Most antibodies showed higher maximal binding than the analogs of the reference antibody 5E5, corresponding to the data obtained using MCF7 cells. Affinity for T47D WT cells was similar to that for MUC1 40 Tn 10 The affinity of the peptides was further closely related.

[0663] Example 7 - Affinity of MUC1 IgG for MC38 cells

[0664] MUC1 IgG was screened for binding to MC38 huMUC1COSMC KO cells stably expressing human MUC1-Tn, MC38 huMUC1 cells expressing normal human MUC1 glycoforms, and MC38 WT cells that do not express human MUC1. An isotype control antibody was included as a negative control antibody, and a reference antibody 5E5 analog was included as a positive control antibody. A reference antibody pankomab analog was included as a control for the relative expression of MUC1 in different cell lines. Other control antibodies included 3F1 and 5F4. MUC1 IgG, isotype controls, 5E5 analogs, and pankomab antibody analogs were titrated using 8-step 4-fold serial dilutions starting at 10ug / ml; 3F1 and 5F4 were used at 10ug / ml.

[0665] MC38 huMUC1 cells (accession number CVCL_5138; Uniprot) were stably transfected with human MUC1 to knock out the mouse COSMC (C1galt1c1) gene (accession number ENSMUST00000058265.7; Ensemble) to generate MC38 huMUC1COSMC KO cells. Cells were placed on ice for 30 minutes with MUC1 IgG and control antibodies diluted in FACS buffer (PBS + 0.5% BSA + 2mM EDTA), washed twice with ice-cold FACS buffer, and stained with anti-human IgG-PE secondary antibody (Invitrogen, catalog number H10104) at 1:100 (for MUC1, isotype control, 5E5 analog, and analogs of pankomab antibody samples), or anti-mouse IgG-PE secondary antibody (Invitrogen, catalog number M30004-4) at 1:400 (for 3F1 and 5F4 antibody samples). Binding was measured using iQue Screener PLUS. The cells were screened using IntelliCyt The MFI values ​​of each HuMUC1-(S)Tn replicate were plotted against IgG concentration and the area under the curve (AUC) was calculated, normalized using isotype antibodies and analogs of the reference antibody 5E5 as negative and positive antibody controls, respectively.

[0666] The results are as follows Figure 6 As shown. Expression of positive and negative control antibodies on MC38 huMUC1 COSMC KO cells, MC38huMUC1 cells, and MC38 WT cells was consistent with expectations. None of the MUC1 antibodies, except for the antibody comprising a heavy chain variable region as shown in SEQ ID NO:178, bound to MC38 WT cells. However, this antibody did not bind to the CALR WT negative control peptide or 293FF WT cells. The plate of MUC1-(S)Tn binders showed a high binding signal to MC38 huMUC1 COSMC KO cells. The plate of MUC1-(S)Tn binders showed a very low binding signal to MC38 huMUC1 cells, demonstrating specificity for MUC1-(S)Tn.

[0667] sequence

[0668] SEQ ID NO: 1—Heavy chain variable region

[0669] QVQLVESGAEVKKPGASVRLSCKASGYSFIHYYLHYVRQAPGQGLEWMGVINPFDGSTDYAQKFHGRVTLTRDTSASTVFMELSSLGSEDTAVYFCTRDNMIGYVLFDYWGLGTLVTVSS

[0670] SEQ ID NO:2 - Heavy chain CDR1

[0671] HYYLH

[0672] SEQ ID NO:3 - Heavy chain CDR2

[0673] VINPFDGSTDYAQKFHG

[0674] SEQ ID NO:4 - Heavy chain CDR3

[0675] DNMIGYVLFDY

[0676] SEQ ID NO:5 - Heavy chain variable region

[0677] QVQLVQSGAEVKKPGASVKVSCKTSGYTFTRYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQKFQGRVTVTRDTSTFTVYMELSSLRSEDTAVYYCARDYMRGYVIFDYWGQGTLVTVSS

[0678] SEQ ID NO:6 - Heavy chain CDR1

[0679] RYYIQ

[0680] SEQ ID NO:7 - Heavy chain CDR2

[0681] VINPYDGSTNYAQKFQG

[0682] SEQ ID NO:8 - Heavy chain CDR3

[0683] DYMRGYVIFDY

[0684] SEQ ID NO:9 - Heavy chain variable region

[0685] QVQLVQSGAEVKKPGASVKVSCKASGYTFNRHYIQWVRQAPGQGLEWMGTINPYDGSTHYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFDYWGQGTLVTVSS

[0686] SEQ ID NO:10—Heavy chain CDR1

[0687] HYIQ

[0688] SEQ ID NO:11—Heavy chain CDR2

[0689] TINPYDGSTHYAQKFQG

[0690] SEQ ID NO:12—Heavy chain CDR3

[0691] DYMIGYVVFDY

[0692] SEQ ID NO:13—Heavy chain variable region

[0693] QVQLVQSGAEVKKPGASLKVSCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQKFQGRVTLTRDTSTTTVYMELSSLRSDDTAVYYCARDYMIGYVVFDYWGQGTLVTVSS

[0694] SEQ ID NO:14—Heavy chain CDR1

[0695] NYYIQ

[0696] SEQ ID NO:15—Heavy chain CDR2

[0697] VINPYDGSTNYAQKFQG

[0698] SEQ ID NO:16—Heavy chain CDR3

[0699] DYMIGYVVFDY

[0700] SEQ ID NO: 17—Heavy chain variable region

[0701] QVQLVQSGAEVKKPGASVRVSCKASGYPFINYYIQWVRQAPGQGLEWMGVVNPYDGSTNYAQKFQGRVSITRDASTSTVFMELNSLRSEDTAVYYCARDSMLGHVIFDFWGQGTLVTVSS

[0702] SEQ ID NO:18—Heavy chain CDR1

[0703] NYYIQ

[0704] SEQ ID NO:19—Heavy chain CDR2

[0705] VVNPYDGSTNYAQKFQG

[0706] SEQ ID NO:20—Heavy chain CDR3

[0707] DSMLGHVIFDF

[0708] SEQ ID NO:21—Heavy chain variable region

[0709] QVQLVQSGAEVRKPGASVKFSCKASGYSFTHYYVHWVRQAPGQGLEWMGVINPFDSSTNYARKFQGRVTMTRDTSTTTVYMDLSSLRSEDTAVYFCARDAMEGDSYFDYWGQGTLVTVSS

[0710] SEQ ID NO:22—Heavy chain CDR1

[0711] HkDJ

[0712] SEQ ID NO:23—Heavy chain CDR2

[0713] VINPFDSSTNYARKFQG

[0714] SEQ ID NO:24—Heavy chain CDR3

[0715] DAMEGDSYFDY

[0716] SEQ ID NO:25—Heavy chain variable region

[0717] QVQLVESGAEVKKPGASVKVSCKASGYTFARYYVHWVRQAPGQGLEWMGLINPYGGATNYAQNFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDAMGGDSYFDYWGQGTLVTVSS

[0718] SEQ ID NO:26—Heavy chain CDR1

[0719] YV

[0720] SEQ ID NO:27—Heavy chain CDR2

[0721] LINPYGGATNYAQNFQG

[0722] SEQ ID NO:28—Heavy chain CDR3

[0723] DAMGGDSYFDY

[0724] SEQ ID NO:29—Heavy chain variable region

[0725] QVQLVQSGAEVKKPGASVKVSCKASGYSFIHYYIHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQDRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDDWGQGTLVTVSS

[0726] SEQ ID NO:30—Heavy chain CDR1

[0727] HYYI

[0728] SEQ ID NO:31—Heavy chain CDR2

[0729] VINPFDGSTNYAQKFQD

[0730] SEQ ID NO:32—Heavy chain CDR3

[0731] DNMIGYVLFDD

[0732] SEQ ID NO:33—Heavy chain variable region

[0733] QVQLVQSGAEVKKPGASVRVSCKASGYPFINYYIQWVRQAPGQGLEWMGVINPFDGSTNYAQKFQDRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLLDYWGQGTLVTVSS

[0734] SEQ ID NO:34—Heavy chain CDR1

[0735] NYYIQ

[0736] SEQ ID NO:35—Heavy chain CDR2

[0737] VINPFDGSTNYAQKFQD

[0738] SEQ ID NO:36—Heavy chain CDR3

[0739] DNMIGYVLLDY

[0740] SEQ ID NO:37—Heavy chain variable region

[0741] QVQLVQSGAEVKKPGASVTVSCKTSGYTFTSYYIHWLRQAPGQGLEWMGVINPFGGATNYAQKFQGRVTMTRDTSTITVYMELSSLRSEDTAVYYCARDAMGGDSHFDHWGRGTLVTVSS

[0742] SEQ ID NO:38—Heavy chain CDR1

[0743] YYI

[0744] SEQ ID NO:39—Heavy chain CDR2

[0745] VINPFGGATNYAQKFQG

[0746] SEQ ID NO:40—Heavy chain CDR3

[0747] DAMGGDSHFDH

[0748] SEQ ID NO:41—Heavy chain variable region

[0749] QVQLVQSGAEVKEPGASMTLSCKTSGYTFISNYIHWLRQAPGQGLEWLGVINPFGGATNYAQKFQGRVTMTRDTSTITVYMELSSLRSEDTAVYYCARDAMGGDSHFDYWGQGTLVTVSS

[0750] SEQ ID NO:42—Heavy chain CDR1

[0751] SNYI

[0752] SEQ ID NO:43—Heavy chain CDR2

[0753] VINPFGGATNYAQKFQG

[0754] SEQ ID NO:44—Heavy chain CDR3

[0755] DAMGGDSHFDY

[0756] SEQ ID NO:45—Heavy chain variable region

[0757] QVQLVESGTEVKKPGASVKVSCKASGYSFIHYYIHWVRQAPGRGLEWMGVINPFDGSTNSAQNFQGRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMLGYVLFDHWGQGTLVTVSS

[0758] SEQ ID NO:46—Heavy chain CDR1

[0759] HYYI

[0760] SEQ ID NO:47—Heavy chain CDR2

[0761] VINPFDGSTNSAQNFQG

[0762] SEQ ID NO:48—Heavy chain CDR3

[0763] DNMLGYVLFDH

[0764] SEQ ID NO:49—Heavy chain variable region

[0765] QVQLVESGAEVKKPGASVKVSCKASGYTFTRYYLHWVRQAPGQGLEWMGVINPYGGATNYAQKFQGRVTLTRDTSTSTVYMELNSLRSEDTAVYFCARDAMEGNSYFDHWGPGTLVTVSS

[0766] SEQ ID NO:50—Heavy chain CDR1

[0767] RYYL

[0768] SEQ ID NO:51—Heavy chain CDR2

[0769] VINPYGGATNYAQKFQG

[0770] SEQ ID NO:52—Heavy chain CDR3

[0771] DAMEGNSYFDH

[0772] SEQ ID NO:53—Heavy chain variable region

[0773] QVQLVQSGAEVKKPGASMTVSCKTSGYTFISYYIHWLRQAPGQGLEWLGVINPFGGATNYAQKFQGRVTMTRDTSTITVYMELSSLRSEDTAVYYCARDAMGGDSHFDFWGQGTLVTVSS

[0774] SEQ ID NO:54—Heavy chain CDR1

[0775] YYI

[0776] SEQ ID NO:55—Heavy chain CDR2

[0777] VINPFGGATNYAQKFQG

[0778] SEQ ID NO:56—Heavy chain CDR3

[0779] DAMGGDSHFDF

[0780] SEQ ID NO:57—Heavy chain variable region

[0781] QVQLVESGTEVKKPGASVKVSCKASGYSFIHYYIHWVRQAPGRGLEWMGVINPFDGSTNYAQNFQGRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDHWGQGTLVTVSS

[0782] SEQ ID NO:58—Heavy chain CDR1

[0783] HYYI

[0784] SEQ ID NO:60—Heavy chain CDR2

[0785] VINPFDGSTNYAQNFQG

[0786] SEQ ID NO:61—Heavy chain CDR3

[0787] DNMIGYVLFDH

[0788] SEQ ID NO:62—Heavy chain variable region

[0789] QVQLVESGAEVKKPGASVKVSCKASGYSFTSYYIQWVRQAPGQGLEWLAVINPIDGSTNYAQKFQGRVTVTRDTSTSTVYMELSSLRSEDTAVYYCARDYMRGFVVFDYWGQGTLVTVSS

[0790] SEQ ID NO:63—Heavy chain CDR1

[0791] Q

[0792] SEQ ID NO:64—Heavy chain CDR2

[0793] VINPIDGSTNYAQKFQG

[0794] SEQ ID NO:65—Heavy chain CDR3

[0795] DYMRGFVVFDY

[0796] SEQ ID NO:66—Heavy chain variable region

[0797] QVQLVQSGAEVKKPGASVRVSCKASGYTFTNYYIHWVRQAPGHGLTWMGVINPFDSSTDYAQKFQGRVTMTRDTSTSTVYMDLSSLRSEDTAVYYCARDYMRGHVVFDYWGQGTLVTVSS

[0798] SEQ ID NO:67—Heavy chain CDR1

[0799] NYYIH

[0800] SEQ ID NO:68—Heavy chain CDR2

[0801] VINPFDSSTDYAQKFQG

[0802] SEQ ID NO:69—Heavy chain CDR3

[0803] DYMRGHVVFDY

[0804] SEQ ID NO:70—Heavy chain variable region

[0805] QVQLVQSGAEVKKPGASVKVSCKASGYAFPNYYIHWVRQAPGQGLEWMGVINPYGGSTNYAQKFQGRVTMTRGTSTSTVYLELSSLRSEDTAVYYCARDAMKGDSFFDYWGQGTLVTVSS

[0806] SEQ ID NO:71—Heavy chain CDR1

[0807] NYYIH

[0808] SEQ ID NO:72—Heavy chain CDR2

[0809] VINPYGGSTNYAQKFQG

[0810] SEQ ID NO:73—Heavy chain CDR3

[0811] DAMKGDSFFDY

[0812] SEQ ID NO:74—Heavy chain variable region

[0813] QVQLVQSGAEVKKPGASVQVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYEQNFQGRVTMTRDTSTNTVYMELNGLRSEDTAVYYCARDHMIGYILFDYWGQGTTVTVSS

[0814] SEQ ID NO:75—Heavy chain CDR1

[0815] NYLQ

[0816] SEQ ID NO:76—Heavy chain CDR2

[0817] TINPFDGSTHYEQNFQG

[0818] SEQ ID NO:77—Heavy chain CDR3

[0819] DHMIGYILFDY

[0820] SEQ ID NO:78—Heavy chain variable region

[0821] QVQLVQSGAEVKKPGTSVKVSCKASGYSFIHYYIHWVRQAPGQGLEWVGVINPFDGSTNYAQKFQGRVTVTRDTSTSAVFMDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS

[0822] SEQ ID NO:79—Heavy chain CDR1

[0823] HYYI

[0824] SEQ ID NO:80—Heavy chain CDR2

[0825] VINPFDGSTNYAQKFQG

[0826] SEQ ID NO:81—Heavy chain CDR3

[0827] DNMIGYVLFDY

[0828] SEQ ID NO:82—Heavy chain variable region

[0829] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQGLEWMGVINPYDSSTNYAQKFQGRVTMTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS

[0830] SEQ ID NO:83—Heavy chain CDR1

[0831] YYI

[0832] SEQ ID NO:84—Heavy chain CDR2

[0833] VINPYDSSTNYAQKFQG

[0834] SEQ ID NO:85—Heavy chain CDR3

[0835] DNMIGYVLFDY

[0836] SEQ ID NO:86—Heavy chain variable region

[0837] QVQLVQSGAEVKKPGASVKVSCKASGYPFINYYIQWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS

[0838] SEQ ID NO:87—Heavy chain CDR1

[0839] NYYIQ

[0840] SEQ ID NO:88—Heavy chain CDR2

[0841] VINPFDGSTNYAQKFQG

[0842] SEQ ID NO:89—Heavy chain CDR3

[0843] DNMIGYVLFDY

[0844] SEQ ID NO:90—Heavy chain variable region

[0845] QVQLVQSGAEVKKPGASVKVSCKASGYTFINHYIHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFLDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS

[0846] SEQ ID NO:91—Heavy chain CDR1

[0847] NHYIH

[0848] SEQ ID NO:92—Heavy chain CDR2

[0849] VINPFDGSTNYAQKFQG

[0850] SEQ ID NO:93—Heavy chain CDR3

[0851] DNMIGYVLFDY

[0852] SEQ ID NO:94—Heavy chain variable region

[0853] QVQLVQSGAEVKKPGSSVKVSCKASGYTFNSYYIHWVRQAPGQGLTWMGVINPFDGSTDYAQKFQGRVTMTRDTSTSTVYMDLSSLRSEDTAVYYCARDYMRGYVVFDYWGRGTLVTVSS

[0854] SEQ ID NO:95—Heavy chain CDR1

[0855] YYI

[0856] SEQ ID NO:96—Heavy chain CDR2

[0857] VINPFDGSTDYAQKFQG

[0858] SEQ ID NO:97—Heavy chain CDR3

[0859] DYMRGYVVFDY

[0860] SEQ ID NO:98—Heavy chain variable region

[0861] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIQWLRQAPGQGLEWMGVINPTGGATNYAQKFQGRVTMTRETSTSMVHMELSSLRSEDTAVYYCARDYMRGYVVFDYWGQGTLVTVSS

[0862] SEQ ID NO:99—Heavy chain CDR1

[0863] Q

[0864] SEQ ID NO: 100—Heavy chain CDR2

[0865] VINPTGGATNYAQKFQG

[0866] SEQ ID NO: 101—Heavy chain CDR3

[0867] DYMRGYVVFDY

[0868] SEQ ID NO: 102—Heavy chain variable region

[0869] QVQLVQSGAEVKKSGASVKVSCKASGYPFISYYIHWLRQAPGQGLTWMGVINPFDGSTDYAQKFQGRVTVTRDTSTSTVYMDLSSLRADDTAIYYCARDYMRGYVVFDYWGQGTLVTVSS

[0870] SEQ ID NO: 103—Heavy chain CDR1

[0871] YYI

[0872] SEQ ID NO: 104—Heavy chain CDR2

[0873] VINPFDGSTDYAQKFQG

[0874] SEQ ID NO: 105—Heavy chain CDR3

[0875] DYMRGYVVFDY

[0876] SEQ ID NO: 106—Heavy chain variable region

[0877] QVQLVQSGAEMKKPGASVKISCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQRFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFDFWGQGTLVTVSS

[0878] SEQ ID NO: 107—Heavy chain CDR1

[0879] NYYIQ

[0880] SEQ ID NO: 108—Heavy chain CDR2

[0881] VINPYDGSTNYAQRFQG

[0882] SEQ ID NO: 109—Heavy chain CDR3

[0883] DYMIGYVVFDF

[0884] SEQ ID NO: 110—Heavy chain variable region

[0885] QVQLVQSGAEMKKPGASVKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGVINPYDGSTNYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFDFWGQGTLVTVSS

[0886] SEQ ID NO:111 - Heavy chain CDR1

[0887] NYYIH

[0888] SEQ ID NO:112 - Heavy chain CDR2

[0889] VINPYDGSTNYAQKFQG

[0890] SEQ ID NO:113 - Heavy chain CDR3

[0891] DYMIGYVVFDF

[0892] SEQ ID NO:114 - Heavy chain variable region

[0893] QVQLVQSGAEMKKPGASVTVSCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQRFQGRVTLTRDTTTTTVNMELSGLRSEDTAVYYCARDYMIGYVVFDFWGQGTLVTVSS

[0894] SEQ ID NO:115 - Heavy chain CDR1

[0895] NYYIQ

[0896] SEQ ID NO:116 - Heavy chain CDR2

[0897] VINPYDGSTNYAQRFQG

[0898] SEQ ID NO:117 - Heavy chain CDR3

[0899] DYMIGYVVFDF

[0900] SEQ ID NO:118 - Heavy chain variable region

[0901] QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQRFQGKITLTRDTSTTTVYMELSSLGSEDTAVYYCARDYMIGYVVFDYWGQGTMVTVSS

[0902] SEQ ID NO: 119—Heavy chain CDR1

[0903] NYYIQ

[0904] SEQ ID NO: 120—Heavy chain CDR2

[0905] VINPYDGSTNYAQRFQG

[0906] SEQ ID NO: 121—Heavy chain CDR3

[0907] DYMIGYVVFDY

[0908] SEQ ID NO:122—Heavy chain variable region

[0909] EVQLLESGAEVKKPGASVKVSCKASGYTFNRYYIHWVRQAPGQGLEWMGVINPYDGSTDYAQKFRDRITMTRDTSTNTVYMDLSSLRSEDTAVYFCARDAMEGDSYFDNWGRGTLVTVSS

[0910] SEQ ID NO:123—Heavy chain CDR1

[0911] YYI

[0912] SEQ ID NO: 124—Heavy chain CDR2

[0913] VINPYDGSTDYAQKFRD

[0914] SEQ ID NO: 125—Heavy chain CDR3

[0915] DAMEGDSYFDN

[0916] SEQ ID NO: 126—Heavy chain variable region

[0917] QVQLVQSGAEVKKPGASVKVSCKASGYTFNRYYLHWVRQAPGQGLEWMGVINPYDGSTNYAQKFRGRVTMTRDTSTNTVYMELSSLRSEDTAVYFCARDAMEGDSYFDNWGQGTTVTVSS

[0918] SEQ ID NO: 127—Heavy chain CDR1

[0919] RYYL

[0920] SEQ ID NO: 128—Heavy chain CDR2

[0921] VINPYDGSTNYAQKFRG

[0922] SEQ ID NO: 129—Heavy chain CDR3

[0923] DAMEGDSYFDN

[0924] SEQ ID NO: 130—Heavy chain variable region

[0925] QVQLVQSGAEVKKPGASVRVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYAQKFQGRVTLTRDTSTSTVYMELNSLRSDDTAVYYCARDYMIGYVLFDYWGQGTLVTVSS

[0926] SEQ ID NO: 131—Heavy chain CDR1

[0927] NYLQ

[0928] SEQ ID NO: 132—Heavy chain CDR2

[0929] TINPFDGSTHYAQKFQG

[0930] SEQ ID NO: 133—Heavy chain CDR3

[0931] DYMIGYVLFDY

[0932] SEQ ID NO: 134—Heavy chain variable region

[0933] QVQLVQSGAEVKKPGASVEVSCKASGYTFINYYIHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFLDLSSLRPEDTAVYYCARDSMIGYVLFDYWGQGTLVTVSS

[0934] SEQ ID NO: 135—Heavy chain CDR1

[0935] NYYIH

[0936] SEQ ID NO: 136—Heavy chain CDR2

[0937] VINPFDGSTNYAQKFQG

[0938] SEQ ID NO: 137—Heavy chain CDR3

[0939] DSMIGYVLFDY

[0940] SEQ ID NO: 138 - Heavy chain variable region

[0941] QVQLVQSGAEVKKPGASVRVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSMIGYVLFDYWGQGTLVTVSS

[0942] SEQ ID NO: 139—Heavy chain CDR1

[0943] NYLQ

[0944] SEQ ID NO: 140—Heavy chain CDR2

[0945] TINPFDGSTHYAQKFQG

[0946] SEQ ID NO: 141 - Heavy chain CDR3

[0947] DSMIGYVLFDY

[0948] SEQ ID NO: 142—Heavy chain variable region

[0949] QVQLVQSGAEVKKPGASVMLSKASGYIFTSYYIQWVRQAPGQGLEWMGVVNPYDGSTNYAQKFQGRVSITRDTSTSTVFMELNSLRSEDTAVYYCARDSMLGHVIFDFWGQGTLVTVSS

[0950] SEQ ID NO: 143—Heavy chain CDR1

[0951] Q

[0952] SEQ ID NO: 144—Heavy chain CDR2

[0953] VVNPYDGSTNYAQKFQG

[0954] SEQ ID NO: 145—Heavy chain CDR3

[0955] DSMLGHVIFDF

[0956] SEQ ID NO: 146—Heavy chain variable region

[0957] QVQLVQSGAEVKKPGASVKVSCKASGYAFTSYYIHWVRQAPGQGLEWMGVINPYGGSTNYAQKFQGRVTMTRDTSTRIVYMDLSSLRSEDTAVYYCARDAMEGDSYFDYWGQGTLVTVSS

[0958] SEQ ID NO:147 - Heavy chain CDR1

[0959] SYYIH

[0960] SEQ ID NO:148 - Heavy chain CDR2

[0961] VINPYGGSTNYAQKFQG

[0962] SEQ ID NO:149 - Heavy chain CDR3

[0963] DAMEGDSYFDY

[0964] SEQ ID NO:150 - Heavy chain variable region

[0965] QVQLVQSGAEVKTPGASVKVSCKASGYTFTNYYIQWVRQAPGQALEWMGVINPYDGSTHYAQKFQGRVTLTRDTSTTTVYMDLSSLRSEDAAVYYCARDYMLGYVVFDYWGQGTLVTVSS

[0966] SEQ ID NO:151 - Heavy chain CDR1

[0967] NYYIQ

[0968] SEQ ID NO:152 - Heavy chain CDR2

[0969] VINPYDGSTHYAQKFQG

[0970] SEQ ID NO:153 - Heavy chain CDR3

[0971] DYMLGYVVFDY

[0972] SEQ ID NO:154 - Heavy chain variable region

[0973] QVQLVQSGAEVKKSGASVKVSCKASGYIFTSYYIQWVRQAPGQGLEWMGVINPFDGSTDYAQKFQGRVTVTRDTSTTTVFMELSSLRSEDTAVYYCARDNMLGYVLFDYWGQGTLVTVSS

[0974] SEQ ID NO: 155—Heavy chain CDR1

[0975] Q

[0976] SEQ ID NO: 156—Heavy chain CDR2

[0977] VINPFDGSTDYAQKFQG

[0978] SEQ ID NO: 157—Heavy chain CDR3

[0979] DNMLGYVLFDY

[0980] SEQ ID NO: 158—Heavy chain variable region

[0981] QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTATTTVFMDLSSLRSEDTAVYYCARDNMLGYVLFDYWGQGTLVTVSS

[0982] SEQ ID NO: 159—Heavy chain CDR1

[0983] YY

[0984] SEQ ID NO: 160—Heavy chain CDR2

[0985] VINPFDGSTNYAQKFQG

[0986] SEQ ID NO: 161—Heavy chain CDR3

[0987] DNMLGYVLFDY

[0988] SEQ ID NO: 162—Heavy chain variable region

[0989] QVQLVQSGAEVKKPGASVMLSKASGYIFTSYYIQWVRQAPGQGLEWMGVVNPYDGSTNYAQKFQGRVTITRDTSTSTVFMELSSLRSEDTAVYYCARDNMLGHVIFDFWGQGTLVTVSS

[0990] SEQ ID NO: 163—Heavy chain CDR1

[0991] Q

[0992] SEQ ID NO: 164—Heavy chain CDR2

[0993] VVNPYDGSTNYAQKFQG

[0994] SEQ ID NO: 165—Heavy chain CDR3

[0995] DNMLGHVIFDF

[0996] SEQ ID NO: 166—Heavy chain variable region

[0997] QVQLVESGAEVKKPGASVKVSCKASGYTFNRYYIHWVRQAPGQGLEWMGVINPSDGSTNYAQKFRGRITMTRDTSTNIVYMELSSLRSEDTAVYFCARDAMGGDSYFDNWGQGTLVTVSS

[0998] SEQ ID NO: 167—Heavy chain CDR1

[0999] YYI

[1000] SEQ ID NO: 168—Heavy chain CDR2

[1001] VINPSDGSTNYAQKFRG

[1002] SEQ ID NO: 169—Heavy chain CDR3

[1003] DAMGGDSYFDN

[1004] SEQ ID NO: 170—Heavy chain variable region

[1005] QVQLVQSGAEVKKPGASVQVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYAQKFQGRVTMTRDTSTNTVYMELNSLRSEDTAVYYCARDRMIGYVLFDYWGQGTLVTVSS

[1006] SEQ ID NO: 171—Heavy chain CDR1

[1007] NYLQ

[1008] SEQ ID NO: 172—Heavy chain CDR2

[1009] TINPFDGSTHYAQKFQG

[1010] SEQ ID NO: 173—Heavy chain CDR3

[1011] DRMIGYVLFDY

[1012] SEQ ID NO: 174—Heavy chain variable region

[1013] EVQLVESGAEVKKPGASVKVSCKASGYTFNRYYLHWVRQAPGQGLEWMGVINPYDGSTNYAQKFRGRITMTRDTSTNTVYMELSSLRSEDTAVYFCARDAMEGDSYFDKWGQGTLVTVSS

[1014] SEQ ID NO: 175—Heavy chain CDR1

[1015] RYYL

[1016] SEQ ID NO: 176—Heavy chain CDR2

[1017] VINPYDGSTNYAQKFRG

[1018] SEQ ID NO: 177—Heavy chain CDR3

[1019] DAMEGDSYFDK

[1020] SEQ ID NO: 178—Heavy chain variable region

[1021] QVQLVQSGAEVKKPGASVKVSCKASGYTFNRHYIQWVRQAPGQGLEWMGTINPYDGSTFYAQKFQDRVTMTRDTSTSTVYMDLNSLKSEDTAVYYCARDYMRGHVLFDYWGQGTLVTVSS

[1022] SEQ ID NO: 179—Heavy chain CDR1

[1023] HYIQ

[1024] SEQ ID NO: 180—Heavy chain CDR2

[1025] TINPYDGSTFYAQKFQD

[1026] SEQ ID NO: 181 - Heavy chain CDR3

[1027] DYMRGHVLFDY

[1028] SEQ ID NO: 182—Heavy chain variable region

[1029] QVQLVQSGAEVKKPGASVKVSCKASGYSFTYSYIQWVRLAPGRGLEWMGVINPYDGSTDYAQKFQGRVTLTRDTSTTTVYMELSSLTSEDTAVYYCARDFMIGYVVFDYWGQGTLVTVSS

[1030] SEQ ID NO: 183—Heavy chain CDR1

[1031] YQ

[1032] SEQ ID NO: 184—Heavy chain CDR2

[1033] VINPYDGSTDYAQKFQG

[1034] SEQ ID NO: 185—Heavy chain CDR3

[1035] DFMIGYVVFDY

[1036] SEQ ID NO: 186—Heavy chain variable region

[1037] QVQLVQSGAEVKKPGASVKVSCKASGYTFNRYYIHWVRQAPGQGLEWMGVINPYDGSTNYAQKFRGRITMTRDTSTNTVYMDLSSLRSEDTAVYFCARDAMEGDSYLDNWGRGTLVTVSS

[1038] SEQ ID NO: 187—Heavy chain CDR1

[1039] YYI

[1040] SEQ ID NO: 188—Heavy chain CDR2

[1041] VINPYDGSTNYAQKFRG

[1042] SEQ ID NO: 189—Heavy chain CDR3

[1043] DAMEGDSYLDN

[1044] SEQ ID NO: 190—Heavy chain variable region

[1045] QVQLVQSGAEVKKPGASVKVSCKASGYAFTNYYIHWVRQAPGHGLTWMGVINPFDSSTDFAQKFQGRLTMTRDTSTSTVYMDLSSLRSEDTAVYYCARDYMRGHVVFDNWGQGTLVTVSS

[1046] SEQ ID NO: 191—Heavy chain CDR1

[1047] NYYIH

[1048] SEQ ID NO: 192—Heavy chain CDR2

[1049] VINPFDSSTDFAQKFQG

[1050] SEQ ID NO: 193—Heavy chain CDR3

[1051] DYMRGHVVFDN

[1052] SEQ ID NO: 194—Heavy chain variable region

[1053] QVQLVQSGAEVKKPGTSVKVSCTASGYTFTNYYIQWVRQAPGQGLEWLGVINPYDGSTDYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMLGYVVFDFWGQGTLVTVSS

[1054] SEQ ID NO: 195—Heavy chain CDR1

[1055] NYYIQ

[1056] SEQ ID NO: 196—Heavy chain CDR2

[1057] VINPYDGSTDYAQKFQG

[1058] SEQ ID NO: 197—Heavy chain CDR3

[1059] DYMLGYVVFDF

[1060] SEQ ID NO: 198—Heavy chain variable region

[1061] QVQLVQSGAEVKKPGASVKVSCKASGYTFINNYIQWVRQAPGQGLEWMGVINPYDGSIHYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFEYWGQGTLVTVSS

[1062] SEQ ID NO: 199—Heavy chain CDR1

[1063] NNYIQ

[1064] SEQ ID NO:200—Heavy chain CDR2

[1065] VINPYDGSIHYAQKFQG

[1066] SEQ ID NO:201—Heavy chain CDR3

[1067] DYMIGYVVFEY

[1068] SEQ ID NO:202—Heavy chain variable region

[1069] QVQLVQSGAEVKNPGASVKFSCKASGYTFINYYLQWLRQAPGQGLEWMGTINPFDGSTSYAQNFQGRVTMTRDTSSTTVYMELNSLKSEDTAVYYCARDSMIGYVLFDSWGQGTLVTVSS

[1070] SEQ ID NO:203—Heavy chain CDR1

[1071] NYLQ

[1072] SEQ ID NO:204—Heavy chain CDR2

[1073] TINPFDGSTSYAQNFQG

[1074] SEQ ID NO:205—Heavy chain CDR3

[1075] DSMIGYVLFDS

[1076] SEQ ID NO:206—Heavy chain variable region

[1077] QVQLVQSGAEVKKPGASVRVSCKASGYPFINYYIQWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFMDLNSLRSEDTAVYYCARDSMIGYVLFDFWGQGTLVTVSS

[1078] SEQ ID NO:207—Heavy chain CDR1

[1079] NYYIQ

[1080] SEQ ID NO:208—Heavy chain CDR2

[1081] VINPFDGSTNYAQKFQG

[1082] SEQ ID NO:209—Heavy chain CDR3

[1083] DSMIGYVLFDF

[1084] SEQ ID NO: 210 - Heavy chain 5E5 analog

[1085] QVQLVQSGAEVKKTGSSVKVSCKASGYTFTDHAIHWVRQAPGQALEWMGHFSPGNTDIKYNDKFKGRVTLTVDRSMSTAYMELSSLRSEDTAMYYCKTSTFFFDYWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1086] SEQ ID NO: 211 - Light chain 5E5 analog

[1087] DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGDQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYSYPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1088] SEQ ID NO: 212 - Heavy chain pankomab analog

[1089] EVKLVESGGGLVQPGGSMKLSCVASGFTFSNYWMNWVRQSPEKGLEWVAEIRLKSNNYTTHYAESVKGRFTISRDDSKSSVSLQMNNLRVEDTGIYYCTRHYYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1090] SEQ ID NO:213 - Light chain pankomab analog

[1091] DIVMTQAAFSNPVTLGTSASISCRSSKSLLHSNGITYFFWYLQKPGLSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLELPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1092] SEQ ID NO:214 - Heavy chain HT186 - D11 analog

[1093] QMQLVQSEAELKKPGASVKVSCKASGYSFTGHYMHWVRQAPGQGLEWMGWIDPVTGGTKYAQNFQGWVTMTRDTSIRTAYLELSRLRSDDTAMYYCAREVTGDRGQFDKWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1094] SEQ ID NO:215 - Light chain HT186 - D11 analog

[1095] QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPALVIYYGSNRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDWVFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1096] SEQ ID NO:216 - Heavy chain isotype control IgG

[1097] EVQLVETGAEVKKPGASVKVSCKASDYIFTKYDINWVRQAPGQGLEWMGWMSANTGNTGYAQKFQGRVTMTRDTSINTAYMELSSLTSGDTAVYFCARSSLFKTETAPYYHFALDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[1098] SEQ ID NO:217 - Light chain isotype control IgG

[1099] DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1100] SEQ ID NO:218 - Light chain variable region

[1101] DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIK

[1102] SEQ ID NO:219 - Light chain CDR1 according to IMGT

[1103] QSISSY

[1104] SEQ ID NO:220 - Light chain CDR2 according to IMGT

[1105] AAS

[1106] SEQ ID NO:221 - Light chain CDR3 according to IMGT

[1107] QQSYSTPPT

[1108] SEQ ID NO:222 - Hinge region

[1109] EPKSCDKTHTCPPCP

[1110] SEQ ID NO:223 - CH1 region

[1111] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRV

[1112] SEQ ID NO:224 - CH2 region

[1113] APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK

[1114] SEQ ID NO:225 - CH3 region

[1115] GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG

[1116] SEQ ID NO:226 - CL region

[1117] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[1118] SEQ ID NO:227 - V region VK1-39

[1119] DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYA ASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTP

[1120] SEQ ID NO:228 - VK1-39 / JK1

[1121] DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIK

[1122] SEQ ID NO:229 - VK1-39 / JK5

[1123] DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK

[1124] SEQ ID NO:230 - Light chain CDR1 according to IMGT

[1125] QSISSY

[1126] SEQ ID NO:231 - Light chain CDR2 according to IMGT

[1127] AAS

[1128] SEQ ID NO:232 - Light chain CDR3 according to IMGT

[1129] QQSYSTPPIT

[1130] SEQ ID NO:233 - V region VK3-15

[1131] EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYG ASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWP

[1132] SEQ ID NO:234 - VK3-15 / JK1

[1133] EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPWTFGQGTKVEIK

[1134] SEQ ID NO:235—Light chain CDR1 according to IMGT

[1135] QSVSSN

[1136] SEQ ID NO:236—Light chain CDR2 according to IMGT

[1137] GAS

[1138] SEQ ID NO:237—Light chain CDR3 according to IMGT

[1139] QQYNNWPWT

[1140] SEQ ID NO:238—V region VK3-20

[1141] EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIY GASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSP

[1142] SEQ ID NO:239-VK3-20 / JK1

[1143] EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK

[1144] SEQ ID NO:240—Light chain CDR1 according to IMGT

[1145] QSVSSSY

[1146] SEQ ID NO:241—Light chain CDR2 according to IMGT

[1147] GAS

[1148] SEQ ID NO:242—Light chain CDR3 according to IMGT

[1149] QYGSSPWT

[1150] SEQ ID NO:243—V region VL3-21

[1151] SYVLTQPPSVSVAPGETARITCGGDNIGRKSVYWYQQKSGQAPVLVIYY DSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDGSSDH

[1152] SEQ ID NO:244-VL3-21 / JL3

[1153] SYVLTQPPSVSVAPGETARITCGGDNIGRKSVYWYQQKSGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDGSSDHWVFGGGTKLTVL

[1154] SEQ ID NO:245—Light chain CDR1 according to IMGT

[1155] NIGRKS

[1156] SEQ ID NO:246—Light chain CDR2 according to IMGT

[1157] YDS

[1158] SEQ ID NO:247—Light chain CDR3 according to IMGT

[1159] QVWDGSSDHWV

[1160] SEQ ID NO:248—Light chain CDR3 according to IMGT

[1161] QQSYSTPPT

[1162] SEQ ID NO:249

[1163] PAPGSTAPPAHGVTSAPDTRPAPG

Claims

1. A binding domain that binds to a MUC1 peptide, wherein the MUC1 peptide comprises an amino acid sequence as shown in SEQ ID NO: 249, wherein the threonine (T) residue at position 14 of the MUC1 peptide comprises α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation. 2 . The binding domain of claim 1 , wherein additionally, the serine (S) residue at position 15 of the MUCl peptide comprises α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

3. The binding domain of claim 1 or 2, which does not bind to a MUC1 peptide comprising the amino acid sequence as shown in SEQ ID NO: 249 but does not comprise α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

4. The binding domain of any one of claims 1 to 3, wherein binding to the MUC-1 peptide is determined in a glycopeptide array.

5. A binding domain that specifically binds to MUC-1 associated with cancer, wherein the binding domain binds to an epitope on MUC1, the epitope comprising a VTSA motif of the N-terminal domain of MUC1, wherein the threonine (T) residue is α-GalNAc glycosylated or Neu5Ac-α(2-6)-GalNAc glycosylated.

6. The binding domain of claim 5, wherein additionally, the serine (S) residue is α-GalNAc glycosylated or Neu5Ac-α(2-6)-GalNAc glycosylated.

7. The binding domain of claim 5 or 6, wherein binding to an epitope on MUC-1 is determined in a glycopeptide array.

8. A binding domain having binding specificity for MUCl associated with cancer, wherein the binding domain comprises a heavy chain variable region comprising: a) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3 and SEQ ID NO: 4, respectively; b) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8, respectively; c) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; d) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 14, SEQ ID NO: 15 and SEQ ID NO: 16, respectively; e) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20, respectively; f) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 22, SEQ ID NO: 23 and SEQ ID NO: 24, respectively; g) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 26, SEQ ID NO: 27 and SEQ ID NO: 28, respectively; h) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 30, SEQ ID NO: 31 and SEQ ID NO: 32, respectively; i) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36, respectively; j) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 38, SEQ ID NO: 39 and SEQ ID NO: 40, respectively; k) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:42, SEQ ID NO:43 and SEQ ID NO:44, respectively; 1) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:46, SEQ ID NO:47 and SEQ ID NO:48, respectively; m) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 51 and SEQ ID NO: 52, respectively; n) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:54, SEQ ID NO:55 and SEQ ID NO:56, respectively; o) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:58, SEQ ID NO:60 and SEQ ID NO:61, respectively; p) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 63, SEQ ID NO: 64 and SEQ ID NO: 65, respectively; q) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:67, SEQ ID NO:68 and SEQ ID NO:69, respectively; r) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:73, respectively; s) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:75, SEQ ID NO:76 and SEQ ID NO:77, respectively; t) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:79, SEQ ID NO:80 and SEQ ID NO:81, respectively; u) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:83, SEQ ID NO:84 and SEQ ID NO:85, respectively; v) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:87, SEQ ID NO:88 and SEQ ID NO:89, respectively; w) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:91, SEQ ID NO:92 and SEQ ID NO:93, respectively; x) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:95, SEQ ID NO:96 and SEQ ID NO:97, respectively; y) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO:99, SEQ ID NO:100 and SEQ ID NO:101, respectively; z) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 103, SEQ ID NO: 104 and SEQ ID NO: 105, respectively; aa) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 107, SEQ ID NO: 108 and SEQ ID NO: 109, respectively; bb) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 111, SEQ ID NO: 112 and SEQ ID NO: 113, respectively; cc) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 115, SEQ ID NO: 116 and SEQ ID NO: 117, respectively; dd) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 119, SEQ ID NO: 120 and SEQ ID NO: 121, respectively; ee) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 123, SEQ ID NO: 124 and SEQ ID NO: 125, respectively; ff) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128 and SEQ ID NO: 129, respectively; gg) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 131, SEQ ID NO: 132 and SEQ ID NO: 133, respectively; hh) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 135, SEQ ID NO: 136 and SEQ ID NO: 137, respectively; ii) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140 and SEQ ID NO: 141, respectively; jj) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 143, SEQ ID NO: 144 and SEQ ID NO: 145, respectively; kk) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 147, SEQ ID NO: 148 and SEQ ID NO: 149, respectively; 11) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 151, SEQ ID NO: 152 and SEQ ID NO: 153, respectively; mm) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 155, SEQ ID NO: 156 and SEQ ID NO: 157, respectively; nn) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 159, SEQ ID NO: 160 and SEQ ID NO: 161, respectively; oo) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 163, SEQ ID NO: 164 and SEQ ID NO: 165, respectively; pp) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 167, SEQ ID NO: 168 and SEQ ID NO: 169, respectively; qq) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 171, SEQ ID NO: 172 and SEQ ID NO: 173, respectively; rr) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 175, SEQ ID NO: 176 and SEQ ID NO: 177, respectively; ss) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 179, SEQ ID NO: 180 and SEQ ID NO: 181, respectively; tt) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 183, SEQ ID NO: 184 and SEQ ID NO: 185, respectively; uu) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188 and SEQ ID NO: 189, respectively; vv) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 191, SEQ ID NO: 192 and SEQ ID NO: 193, respectively; ww) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 195, SEQ ID NO: 196 and SEQ ID NO: 197, respectively; xx) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) having the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200 and SEQ ID NO: 201, respectively; yy) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 203, SEQ ID NO: 204 and SEQ ID NO: 205, respectively; or zz) heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2) and heavy chain CDR3 (HCDR3) have the amino acid sequences shown in SEQ ID NO: 207, SEQ ID NO: 208 and SEQ ID NO: 209, respectively; Each HCDR may contain up to three, two or one amino acid substitution.

9. The binding domain of claim 8, wherein the binding domain comprises a heavy chain variable region having an amino acid sequence as shown in any one of SEQ ID NO: 1; 5; 9; 13; 17; 21; 25; 29; 33; 37; 41; 45; 49; 53; 57; 62; 66; 70; 74; 78; 82; 86; 90; 94; 98; 102; 106; 110; 114; 118; 122; 126; 130; 134; 138; 142; 146; 150; 154; 158; 162; 166; 170; 174; 178; 182; 186; 190; 194; 198; 202 or 206, or having a sequence identity of at least 80%, or 85%, or 90%, or 95% thereof.

10. The binding domain of claim 8 or 9, wherein the binding domain comprises a light chain variable region comprising a light chain CDR1 (LCDR1), a light chain CDR2 (LCDR2) and a light chain CDR3 (LCDR3), having the amino acid sequences shown in SEQ ID NO: 219, SEQ ID NO: 220 and SEQ ID NO: 221, respectively, wherein each LCDR may contain up to three, two or one amino acid substitutions.

11. The binding domain of any one of claims 8 to 10, wherein the binding domain comprises a light chain variable region having an amino acid sequence as shown in SEQ ID NO: 218, or having a sequence identity of at least 80%, or 85%, or 90%, or 95% thereto.

12. The binding domain of any one of claims 1 to 11, wherein the binding domain is a Fab.

13. A binding moiety comprising two binding domains according to any one of claims 1 to 12.

14. The binding moiety of claim 13, wherein the binding moiety is an IgG1 or IgG4 antibody.

15. A binding moiety according to claim 13 or 14, wherein binding of the binding moiety to an Fc receptor is abolished or reduced, or wherein binding of the binding moiety to an Fc receptor is enhanced.

16. The binding moiety according to any one of claims 13 to 15, wherein the binding moiety exhibits a higher binding signal than a reference antibody, the reference antibody comprising two heavy chains having the amino acid sequence shown in SEQ ID NO: 210 and a light chain having the amino acid sequence shown in SEQ ID NO:

211.

17. A pharmaceutical composition comprising an effective amount of a binding domain according to any one of claims 1 to 12, or a binding moiety according to any one of claims 13 to 16, and a pharmaceutically acceptable carrier.

18. A binding domain according to any one of claims 1 to 12, a binding moiety according to any one of claims 13 to 16, or a pharmaceutical composition according to claim 17 for use in therapy, in particular for the treatment of cancer.

19. A method for treating a disease, in particular for treating cancer, comprising administering to an individual in need thereof an effective amount of a binding domain according to any one of claims 1 to 12, a binding moiety according to any one of claims 13 to 16, or a pharmaceutical composition according to claim 17.

20. A nucleic acid comprising a nucleic acid sequence encoding a heavy chain variable region as defined in claim 8 or 9, in particular wherein the nucleic acid further comprises a nucleic acid sequence encoding a CH1 region and optionally a hinge region, a CH2 region and a CH3 region.

21. A cell comprising a nucleic acid sequence encoding a heavy chain variable region as defined in claim 8 or 9, in particular wherein the cell further comprises a nucleic acid sequence encoding a CH1 region and optionally a hinge region, a CH2 region and a CH3 region.

22. The cell according to claim 21, wherein the cell further comprises at least one nucleic acid sequence encoding a light chain variable region, in particular wherein the light chain variable region comprises a light chain variable region as defined in claim 10 or 11 and optionally a CL region.

23. A cell which produces a binding domain according to any one of claims 1 to 12, or a binding moiety according to any one of claims 13 to 16.

24. A kit comprising a container comprising a binding domain according to any one of claims 1 to 12, or a binding moiety according to any one of claims 13 to 16, and optionally instructions for use.

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