Binding domain to cancer-related MUC1

Abstract

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

Description

[Technical Field]

[0001] This disclosure relates to binding domains and binding moieties, for example, in the field of antibodies. More specifically, this disclosure relates to the field of therapeutic antibodies for the treatment of cancer. More specifically, this disclosure relates to binding domains that bind to cancer-related MUC1, and binding moieties that include such binding domains. [Background technology]

[0002] Mucin 1, or MUC1, is a transmembrane glycoprotein typically expressed primarily in glandular or luminal epithelial cells. MUC1 forms a physical barrier for lubrication and protection and assists in signal transduction (Shimizu M, Yamauchi KJ Biochem. 1982;91(2):515-24). The extracellular domain of MUC1 can extend from the cell surface to 200–500 nm and is highly glycosylated. Its sugar branches are negatively charged and can be oligomerized, thereby creating a physical barrier that protects the cell (Nath S, Mukherjee P. Trends Mol Med. 2014;20(6):332-42).

[0003] The MUC1 gene is located on chromosome 1:155,185,824-155,192,916 (GRCh38:CM000663.2) and has 29 transcripts. MUC1 is also known under several different aliases, including: mucin 1, cell surface-associated, transmembrane, PEM, PUM, EMA, H23Ag, epicyalin, cancer antigen 15-3, ADMCKD(1), CD227, MCKD(1), MCD, tumor-associated epithelial membrane antigen, cancer-associated mucin, tumor-associated epithelial mucin, and tumor-associated mucin. The external IDs for the MUC1 gene are HGNC:7508 NCBI;Entrez Gene:4582;Ensembl:ENSG00000185499;OMIM:158340 and UniProtKB:P15941.

[0004] The MUC1 protein consists of a single polypeptide chain that is post-translationally cleaved into two peptide subunits called MUC1-C and MUC-N by autoproteolysis at the sea urchin sperm protein enterokinase and agrin (SEA) domains, both of which remain associated via stable non-covalent bonds (Figure 1, Gao T. et al. Biomed Pharmacother. 2020;132:110888). MUC1-C consists of a short extracellular domain with 58 amino acids, a hydrophobic transmembrane domain with 28 amino acids, and an intracellular cytoplasmic domain with 69 amino acids. MUC1-N is located extracellularly and contains approximately 20 amino acid sequence repeats with a repeat number that varies from 20 to 120, designated as a variable-number tandem repeat (VNTR) (HGVTSAPDTRPAPGSTAPPA). VNTR is rich in serine and threonine residues, and each repeat contains five potential sites for O-glycosylation, which, under normal conditions, highly glycosylate MUC1 in the normal pattern of O-glycosylation (Bose M, Mukherjee P. Vaccines (Basel). 2020; 8(4): 659, Zhou D. et al. Molecules. 2018; 23(6): 1326).

[0005] During malignant transformation, MUC1 is overexpressed, losing its polarity and apex distribution, with expression occurring both on the surface and in the cytoplasm of epithelial cells. In normal cells, O-glycosylation progresses to mature, elongated, branched O-glycans, but 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 containing cleaved O-glycans known as Tn antigens (GalNAcα-O-serine / threonine) and their sialylated version, the sialyl-Tn antigen (STn antigen), which contains sialic acid α-2,6 bound to GalNAcα-O-serine / threonine (Figure 1, Gao T. et al. Biomed Pharmacother. 2020).

[0006] The expression of tumor-associated carbohydrates (TACAs) is involved in immunomodulation and inhibition of anti-cancer immune responses, and the expression of sialic acid-containing glycans in tumor cells is correlated with metastatic phenotype and poor prognosis in patients (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 typically expressed at high levels in certain tumor cell tissues, 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).

[0007] The development of therapeutic antibodies against Tn / STn antigens has been reported. Gatipotuzumab (formerly known as PankoMAb-GEX) has been reported to exhibit high affinity and specific binding to cancer cells, but it is no longer being investigated in clinical trials (Danielczyk A. et al. Cancer Immunol Immunother. 2006;55(11):1337-47). Its safety was reported in a Phase I trial (Fiedler W. et al. Eur J Cancer. 2016;63:55-63), however, when applied as monotherapy for maintenance in relapsed ovarian cancer (OC), gatipotuzumab did not show beneficial effects in a Phase IIb trial in advanced ovarian cancer (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 that has been reported to exhibit specificity against cancer cells (Sorensen AL. et al. Glycobiology. 2006;16(2):96-107). Using 5E5, a chimeric antigen receptor (CAR) targeting the TnMUC antigen was generated. This CART-TnMUC1 construct consists of a mouse anti-human scFv derived from the monoclonal antibody 5E5 that recognizes the Tn glycan-containing epitope of MUC1, and is being evaluated in clinical trials for the treatment of refractory solid tumors and malignancies.

[0008] Therefore, there remains a need for novel therapeutic interventions that selectively target cancer-related MUC1s. [Overview of the Initiative]

[0009] In certain embodiments, this disclosure provides novel pharmaceuticals for the treatment of human diseases, specifically, cancer.

[0010] In certain embodiments, the present disclosure relates to a binding domain that binds to a MUC1 peptide, the MUC1 peptide comprising the amino acid sequence PAPGSTAPPAHGVTSAPDTRPAPG shown in SEQ ID NO: 249, wherein the threonine (T) residue at position 14 of the MUC1 peptide is α-GalNAc glycosylated or Neu5Ac-α(2-6)-GalNAc glycosylated.

[0011] In certain embodiments, the present disclosure relates to a binding domain that specifically binds to cancer-related MUC1, the binding domain binding to an epitope comprising the 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.

[0012] In certain embodiments, the present disclosure relates to a binding domain having binding specificity for cancer-related MUC1, the binding domain comprising a heavy chain variable region further defined herein.

[0013] In certain embodiments, the present disclosure relates to a binding moiety comprising two binding domains described herein.

[0014] In certain embodiments, the present disclosure relates to a pharmaceutical composition comprising an effective amount of a binding domain or binding moiety described herein.

[0015] In certain embodiments, the present disclosure relates to a binding domain, binding moiety, or pharmaceutical composition described herein for use in therapy.

[0016] In certain embodiments, the present disclosure relates to a binding domain, binding moiety, or pharmaceutical composition described herein for use in the treatment of cancer.

[0017] In certain embodiments, the present disclosure relates to a method for treating a disease, the method comprising administering to an individual in need of treatment an effective amount of a binding domain, binding moiety, or pharmaceutical composition described herein.

[0018] In certain embodiments, the present disclosure relates to a method for treating cancer, the method comprising administering to an individual in need of treatment an effective amount of a binding domain, binding moiety, or pharmaceutical composition described herein.

[0019] In certain embodiments, the present disclosure relates to a nucleic acid sequence encoding a heavy chain variable region as defined herein.

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

[0021] In certain embodiments, the present disclosure relates to a cell that produces a binding domain or binding moiety described herein.

[0022] In certain embodiments, the present disclosure relates to a kit comprising a container containing a binding domain or binding moiety described herein and, optionally, instructions for use.

Best Mode for Carrying Out the Invention

[0023] In certain embodiments, the Disclosure provides novel pharmaceuticals for the diagnosis and treatment of diseases, specifically for the diagnosis and treatment of diseases in humans, specifically for the diagnosis and treatment of cancer. In certain embodiments, the Disclosure provides binding domains that specifically bind to cancer-associated MUC1 but not specifically to MUC1 expressed on non-cancerous cells. Specifically, in certain embodiments, the Disclosure provides for the first time a genus of binding domains that specifically bind to cancer-associated MUC1, comprising Tn(α-GalNAc) glycosylation and / or STn(Neu5Ac-α(2-6)-GalNAc) glycosylation at a specific amino acid residue or a specific amino acid residue of a VTSA motif, which differs from conventional glycosylated MUC1.

[0024] In a particular embodiment, the Disclosure provides a binding domain that binds to the MUC1 peptide, the MUC1 peptide having the amino acid sequence PAPGSTAPPAHGV T SAPDTRPAPG (SEQ ID NO: 249) comprises or consists of, wherein the threonine (T) residue at position 14 of the peptide, as shown in bold and underlined herein, comprises α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

[0025] In certain embodiments, the Disclosure provides a binding domain that specifically binds to cancer-associated MUC-1, the binding domain binding to an epitope containing a VTSA motif in the N-terminal domain of MUC1, the threonine (T) residue being α-GalNAc glycosylated or Neu5Ac-α(2-6)-GalNAc glycosylated.

[0026] In certain embodiments, the binding domain of the Disclosure binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*SAPDTRPAPG (SEQ ID NO: 249), where * represents Tn glycosylation. In certain embodiments, the binding domain of the Disclosure binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*SAPDTRPAPG (SEQ ID NO: 249), where * represents STn glycosylation.

[0027] In certain embodiments, the binding domain of the Disclosure includes Tn or STn glycosylation of the serine (S) residue at position 15 of the MUC1 peptide of SEQ ID NO: 249, in addition to Tn or STn glycosylation of the threonine residue at position 14 of the MUC1 peptide of SEQ ID NO: 249. In certain embodiments, the binding domain of the Disclosure binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*S*APDTRPAPG (SEQ ID NO: 249), where * represents Tn glycosylation. In certain embodiments, the binding domain of the Disclosure binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*S*APDTRPAPG (SEQ ID NO: 249), where * represents STn glycosylation. In certain embodiments, the binding domain of the Disclosure binds to the MUC1 peptide, which comprises the amino acid sequence PAPGSTAPPAHGVT*S ● Contains or consists of APDTRPAPG (SEQ ID NO: 249), where * indicates Tn glycosylation. ● This represents STn glycosylation. In certain embodiments, the binding domain of the present disclosure binds to the MUC1 peptide, which has the amino acid sequence PAPGSTAPPAHGVT*S ● Contains or consists of APDTRPAPG (SEQ ID NO: 249), where * indicates STn glycosylation. ● This represents Tn glycosylation.

[0028] Tn glycosylation refers to the presence of an α-GalNAc group, while STn glycosylation refers to a Neu5Ac-α(2-6)-GalNAc group.

[0029] In certain embodiments, the binding domain of this disclosure binds to human MUC1.

[0030] A “binding domain” refers to a protein molecule, or a portion of a protein molecule, that binds to a target molecule, as can be determined by binding assays well known to those skilled in the art. A binding domain may, for example, be a heavy chain variable region, scFv, or a heavy chain variable region paired with a light chain variable region such as Fab, or a heavy chain variable region linked to a light chain variable region. In certain embodiments, the binding domain of this disclosure is Fab. In certain embodiments, “Fab” means a binding domain comprising a heavy chain variable region and a light chain variable region. In certain embodiments, “Fab” means a binding domain comprising a heavy chain variable region, a light chain variable region, CH1, and a CL region.

[0031] In certain embodiments of this disclosure, the binding domain specifically binds to cancer-related MUC1, meaning that the binding domain does not bind to MUC1 peptides lacking cancer-related aberrant glycosylation patterns at a detectable level, or only at much higher concentrations determined by the glycopeptide array described herein, specifically the glycopeptide array. Examples of cancer-related aberrant glycosylation patterns include Tn glycosylation (α-GalNAc) and STn glycosylation (Neu5Ac-α(2-6)-GalNAc). Therefore, in certain embodiments of this disclosure, the binding domain does not bind to, or substantially does not bind to, MUC1 peptides containing or consisting of the amino acid sequence shown in SEQ ID NO: 249, which does not contain α-GalNAc glycosylation or Neu5Ac-α(2-6)-GalNAc glycosylation.

[0032] In certain embodiments, the binding domain of the Disclosure binds to an epitope containing the VTSA motif of the N-terminal domain of MUC1. In certain embodiments, the binding domain of the Disclosure does not bind to or substantially binds to the MUC1 peptide comprising the amino acid sequence PAPGS*TAPPAHGVTSAPDTRPAPG (SEQ ID NO: 249), where * represents Tn or STn glycosylation. In certain embodiments, the binding domain of the Disclosure does not bind to or substantially binds to the MUC1 peptide comprising the amino acid sequence PAPGS*TAPPAHGVTSAPDTRPAPG (SEQ ID NO: 249), where * represents STn glycosylation. In certain embodiments, the binding domain of the Disclosure does not bind to or substantially binds to the MUC1 peptide comprising the amino acid sequence PAPGST*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), where * represents Tn or STn glycosylation. In certain embodiments, the binding domain of the Disclosure binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGST*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), where * represents Tn glycosylation. In certain embodiments, the binding domain of the Disclosure does not bind to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGST*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249), where * represents STn glycosylation. In certain embodiments, the binding domain of the Disclosure does not bind to or substantially does not bind to the MUC1 peptide comprising or consisting of the amino acid sequence PAPGSTAPPAHGVTS*APDTRPAPG (SEQ ID NO: 249), where * represents Tn or STn glycosylation. In certain embodiments, the binding domain of the present disclosure comprises the amino acid sequence PAPGSTAPPAHGVTS*APDTRPAPG (SEQ ID NO: 249), or does not bind to or substantially does not bind to the MUC1 peptide comprising the same, where * represents Tn glycosylation.In certain embodiments, the binding domain of the Disclosure contains the amino acid sequence PAPGSTAPPAHGVTSAPDT*RPAPG (SEQ ID NO: 249) or does not bind to or substantially binds to the MUC1 peptide consisting of the same, where * represents Tn or STn glycosylation. In certain embodiments, the binding domain of the Disclosure contains the amino acid sequence PAPGSTAPPAHGVTSAPDT*RPAPG (SEQ ID NO: 249) or does not bind to or substantially binds to the MUC1 peptide consisting of the same, where * represents Tn glycosylation. In certain embodiments, the binding domain of the Disclosure contains the amino acid sequence PAPGS*T*APPAHGVTSAPDTRPAPG (SEQ ID NO: 249) or does not bind to or substantially binds to the MUC1 peptide consisting of the same, where * represents Tn or STn glycosylation. In certain embodiments, the binding domain of the Disclosure also binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGST*APPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), where * represents Tn glycosylation. In certain embodiments, the binding domain of the Disclosure does not bind to or substantially binds to the MUC1 peptide comprising or consisting of the amino acid sequence PAPGST*APPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), where * represents STn glycosylation. In certain embodiments, the binding domain of the Disclosure does not bind to or substantially binds to the MUC1 peptide comprising or consisting of the amino acid sequence PAPGS*T*APPAHGVTSAPDT*RPAPG (SEQ ID NO: 249), where * represents Tn or STn glycosylation. In certain embodiments, the binding domain of the present disclosure also binds to the MUC1 peptide, which comprises or consists of the amino acid sequence PAPGSTAPPAHGVT*SAPDT*RPAPG (SEQ ID NO: 249), where * represents Tn or STn glycosylation.

[0033] In certain embodiments, the binding domain is considered to bind to the MUC1 peptide if, in the bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is at least about twice, or about two to three times, or about three times higher than that of the negative isotype control in the glycopeptide array. In certain embodiments, the binding domain is considered to bind to the MUC1 peptide if, in the bivalent monospecific IgG antibody format, it binds to the peptide with an EC50 (ug / ml) of at least less than about 200 or 100 or 50 or 10 or 1 or 0.5 in the glycopeptide array, or about 200 to 0.001 or 100 to 0.001 or 50 to 0.001 or 10 to 0.001 or 1 to 0.001 or 0.5 to 0.001 or 200 to 0.05 or 100 to 0.05 or 50 to 0.05 or 10 to 0.05 or 1 to 0.05 or 0.5 to 0.05. In certain embodiments, the binding domain of the present disclosure binds to the MUC1 peptide if, in a bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is at least about twice, or about two to three times, or about three times higher than that of a negative isotype control in a glycopeptide array. In certain embodiments, the binding domain is considered to bind to the MUC1 peptide if, in a bivalent monospecific IgG antibody format, it binds to the peptide with an EC50 (ug / ml) of at least less than about 200 or 100 or 50 or 10 or 1 or 0.5 in a glycopeptide array, or about 200 to 0.001 or 100 to 0.001 or 50 to 0.001 or 10 to 0.001 or 1 to 0.001 or 0.5 to 0.001 or 200 to 0.05 or 100 to 0.05 or 50 to 0.05 or 10 to 0.05 or 1 to 0.05 or 0.5 to 0.05.In certain embodiments, the binding domain is considered to bind to the MUC1 peptide if, in a bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is at least about twice, or about two to three times, or about three times higher than that of the negative isotype control in the glycopeptide array, and if, in the glycopeptide array, it has an EC50 (ug / ml) of at least less than about 200, 100, 50, 10, 1, or 0.5, or about 200 to 0.001, 100 to 0.001, 50 to 0.001, 10 to 0.001, 1 to 0.001, 0.5 to 0.001, 200 to 0.05, 100 to 0.05, 50 to 0.05, 10 to 0.05, 1 to 0.05, or 0.5 to 0.05. In certain embodiments, the binding domain of the present disclosure binds to the MUC1 peptide in a bivalent monospecific IgG antibody format with a binding signal at least about twice, or about two to three times, or about three times higher than that of a negative isotype control in a glycopeptide array, and has an EC50 (ug / ml) of at least less than about 200, 100, 50, 10, 1, or 0.5 in the glycopeptide array, or about 200 to 0.001, 100 to 0.001, 50 to 0.001, 10 to 0.001, 1 to 0.001, 0.5 to 0.001, 200 to 0.05, 100 to 0.05, 50 to 0.05, 10 to 0.05, 1 to 0.05, or 0.5 to 0.05.

[0034] In certain embodiments, the binding domain is considered not to bind to the MUC1 peptide or substantially not to bind if, in a bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is less than approximately twice as high, approximately two to three times higher, or approximately three times higher than that of the negative isotype control in the glycopeptide array. In certain embodiments, the binding domain is considered not to bind to the MUC1 peptide or substantially not to bind if, in a bivalent monospecific IgG antibody format, it binds to the peptide with an EC50 (ug / ml) higher than approximately 200 in the glycopeptide array. In certain embodiments, the binding domain of the present disclosure is considered not to bind to the MUC1 peptide or substantially not to bind if, in a bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is less than twice as high, approximately two to three times higher, or approximately three times higher than that of the negative isotype control in the glycopeptide array. In certain embodiments, the binding domain is considered not to bind to the MUC1 peptide or substantially not to bind if, in a bivalent monospecific IgG antibody format, it binds to the peptide with an EC50 (ug / ml) higher than 200 in the glycopeptide array. In certain embodiments, the binding domain is considered not to bind to the MUC1 peptide or substantially not to bind if, in the bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is less than approximately twice as high, approximately two to three times as high, or approximately three times higher than the negative isotype control in the glycopeptide array. In certain embodiments, the binding domain is considered not to bind to the MUC1 peptide or substantially not to bind if, in the bivalent monospecific IgG antibody format, it binds to the peptide with an EC50 (ug / ml) higher than approximately 200 in the glycopeptide array. In certain embodiments, the binding domain is considered not to bind to the MUC1 peptide or substantially not to bind if, in the bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is less than approximately twice as high, approximately two to three times as high, or approximately three times higher than the negative isotype control in the glycopeptide array, and has an EC50 (ug / ml) higher than approximately 200 in the glycopeptide array.In certain embodiments, a binding domain is considered not to bind to the MUC1 peptide, or substantially not to bind, if, in a bivalent monospecific IgG antibody format, it binds to the peptide with a binding signal that is less than approximately twice, approximately two to three times, or approximately three times higher than that of the negative isotype control in the glycopeptide array, and if it has an EC50 (ug / ml) greater than 200 in the glycopeptide array.

[0035] For the purposes of this disclosure, the determination of whether a binding domain binds to the MUC1 peptide is made by using a glycopeptide array. The binding data for the binding domains provided herein are obtained by the glycopeptide array described in Example 5. Thus, in certain embodiments, the binding of the binding domain of this disclosure described herein to the MUC1 peptide, or its interaction with the VTSA epitope, is determined by the glycopeptide array described in Example 5.

[0036] In short, the glycopeptide array described in Example 5 is prepared using the O-glycopeptides listed in Table 2. The array is blocked before adding MUC1 IgG and control antibody in the range of 10 μg / ml to 0.6 ng / ml. The array is covered with an adhesive film and shaken at room temperature. The array is then washed and the detection antibody is applied to the array. It is shielded from light and shaken at room temperature for 1 hour, followed by washing. The array is read using an Innopsys InnoScan 710 microarray scanner and suitable software.

[0037] The binding domains or binding moieties of this disclosure bind to cancer-associated MUC1, meaning they do not bind to normal glycosylated MUC1 on non-tumor cells. As used herein, “binding to cancer-associated MUC1” refers to the typical binding ability of a binding domain or binding site to cancer-associated MUC1 present on tumor cells or cell lines engineered to express cancer-associated MUC1. The binding of an antigen-binding domain or binding moiety can be evaluated in various ways. For the purposes of this disclosure, whether a binding domain or binding moiety binds to cancer-associated MUC1 is determined by incubating the binding domain or binding moiety with cells expressing cancer-associated MUC1, such as MC38 huMUC1 COSMC KO cells, T47D huGalNAc cells stably expressing human MUC1-STn, and / or T47D WT cells expressing moderate levels of MUC-Tn, as described herein. Unbound binding domains or binding portions are removed, and bound binding domains or binding portions are detected, for example, by a labeled antibody that binds to the constant domain of the bound binding domain or binding portion.

[0038] Antigen binding can be described in terms of specificity and affinity. Specificity determines which antigen or its epitope is specifically bound by the binding domain or binding moiety. Affinity is a measure of the strength of binding to a particular antigen or epitope.

[0039] In certain embodiments, the binding domain of this disclosure binds to cancer-associated human MUC1.

[0040] In certain embodiments, the combined domain of this disclosure is a) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4. b) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8. c) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12. d) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16. 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), respectively, having the amino acid sequences shown in SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24. g) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 28. h) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 30, SEQ ID NO: 31, and SEQ ID NO: 32. i) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, j) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, k) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 42, SEQ ID NO: 43, and SEQ ID NO: 44. l) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 46, SEQ ID NO: 47, and SEQ ID NO: 48. m) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO: 52. n) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 54, SEQ ID NO: 55, and SEQ ID NO: 56. o) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 58, SEQ ID NO: 60, and SEQ ID NO: 61. p) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65. q) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 67, SEQ ID NO: 68, and SEQ ID NO: 69. r) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 71, SEQ ID NO: 72, and SEQ ID NO: 73. s) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 75, SEQ ID NO: 76, and SEQ ID NO: 77. t) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 79, SEQ ID NO: 80, and SEQ ID NO: 81. u) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 83, SEQ ID NO: 84, and SEQ ID NO: 85. v) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 87, SEQ ID NO: 88, and SEQ ID NO: 89. w) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 91, SEQ ID NO: 92, and SEQ ID NO: 93. x) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 95, SEQ ID NO: 96, and SEQ ID NO: 97. y) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 99, SEQ ID NO: 100, and SEQ ID NO: 101. z) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 103, SEQ ID NO: 104, and SEQ ID NO: 105. aa) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 107, SEQ ID NO: 108, and SEQ ID NO: 109. bb) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 111, SEQ ID NO: 112, and SEQ ID NO: 113. cc) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 115, SEQ ID NO: 116, and SEQ ID NO: 117. dd) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 119, SEQ ID NO: 120, and SEQ ID NO: 121. ee) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 123, SEQ ID NO: 124, and SEQ ID NO: 125. ff) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO: 129. gg) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 131, SEQ ID NO: 132, and SEQ ID NO: 133. hh) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 135, SEQ ID NO: 136, and SEQ ID NO: 137. ii) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140, and SEQ ID NO: 141, jj) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 143, SEQ ID NO: 144, and SEQ ID NO: 145. kk) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 147, SEQ ID NO: 148, and SEQ ID NO: 149. ll) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 151, SEQ ID NO: 152, and SEQ ID NO: 153. (mm) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 155, SEQ ID NO: 156, and SEQ ID NO: 157. nn) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 159, SEQ ID NO: 160, and SEQ ID NO: 161. oo) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 163, SEQ ID NO: 164, and SEQ ID NO: 165. pp) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 167, SEQ ID NO: 168, and SEQ ID NO: 169. qq) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 171, SEQ ID NO: 172, and SEQ ID NO: 173. rr) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 175, SEQ ID NO: 176, and SEQ ID NO: 177. ss) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 179, SEQ ID NO: 180, and SEQ ID NO: 181. tt) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 183, SEQ ID NO: 184, and SEQ ID NO: 185. uu) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188, and SEQ ID NO: 189. vv) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 191, SEQ ID NO: 192, and SEQ ID NO: 193. ww) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 195, SEQ ID NO: 196, and SEQ ID NO: 197. xx) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200, and SEQ ID NO: 201. yy) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 203, SEQ ID NO: 204, and SEQ ID NO: 205, or Each zz) contains a heavy chain variable region comprising heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 207, SEQ ID NO: 208, and SEQ ID NO: 209.

[0041] The heavy chain variable region of the MUC1-binding domain of this disclosure may include a limited number, for example, one, two, or three non-conservative amino acid substitutions, or an unlimited number of conservative amino acid substitutions.

[0042] In certain embodiments, the MUC1-binding domain of the Disclosure also includes MUC1-binding domain variants, and each HCDR may contain up to three, two, or one amino acid variation, e.g., substitutions. In certain embodiments, only one or two HCDRs may contain up to three, two, or one amino acid variation, e.g., substitutions. In certain embodiments, such variants do not contain amino acid variation in HCDR3. In certain embodiments, the amino acid variation is a conservative amino acid substitution.

[0043] Typically, conserved amino acid substitutions involve variations in amino acids that have homologous amino acid residues, which are residues that share similar characteristics or properties. Homologous amino acids are known in the art, as routine methods for performing amino acid substitutions within antibody-binding domains without significantly affecting antibody binding or function are known; for example, see the Lehninger principle of biochemistry (Nelson DL, Cox MM and Lehninger AL. New York: WH Freeman, 2017) or the handbook of biochemistry (Berg JM, Tymoczko JL and Stryer L. New York: WH Freeman, 2007), which are incorporated herein in their entirety. In determining whether an amino acid can be replaced with a conserved amino acid, evaluation may typically be based on, but is not limited to, factors such as (a) the structure of the polypeptide backbone in the substitution region, e.g., sheet or helical structure, (b) the charge or hydrophobicity of the molecule at the target site, and / or (c) the bulk of the side chain. When a residue can be substituted with a residue having a similar side chain or a common characteristic such as similar charge or hydrophobicity, such residues are preferred as substitutions. For example, the following groups can be determined: (1) Nonpolar: Ala(A), Gly(G), Val(V), Leu(L), Ile(I), Pro(P), Phe(F), Trp(W), Met(M), (2) Non-charged: Ser(S), Thr(T), Cys(C), Tyr(Y), Asn(N), Gln(Q), (3) Acidic: Asp(D), Glu(E), and (4) Basic: Lys(K), Arg(R), His(H). Alternatively, amino acids can be grouped as follows: (1) aromatic: Phe(F), Trp(W), Tyr(Y), (2) nonpolar: 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), and (6) polar: Gln(Q), Asn(N), Ser(S), Thr(T), Tyr(Y). Alternatively, amino acid residues may be divided into groups based on 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 that influence chain orientation: Gly (G), Pro (P), and (6) aromatic: Trp (W), Tyr (Y), Phe (F).

[0044] Substitution with another amino acid residue present in the same group is preferred. Therefore, conservative amino acid substitutions may involve exchanging one member of these classes with another member of the same class. Typically, the variation does not result in a loss, or substantially no loss, of the binding specificity of the binding domain to its intended target.

[0045] Additional types of amino acid variations include variations resulting from somatic hypermutation and / or affinity maturation. Binding variants included in this disclosure include somatic hypermutation or affinity-matured heavy chain variable regions that are heavy chain variable regions derived from the same VH gene segment as the heavy chain variable regions sequenced herein, and the variants have amino acid variations including non-conservative and / or conserved amino acid substitutions in one, two, or all three HCDRs. Routine methods for affinity maturation of antibody-binding domains are widely known in the art; see, for example, Tabasinezhad M. et al. Immunol Lett. 2019;212:106-113.

[0046] Whether amino acid residues within the CDR and / or framework region can be substituted, for example, with conserved amino acid residues, and whether such substitution can be done without or substantially without loss of binding specificity, can be determined by methods well known in the art. Examples of experimental methods 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). For example, computational methods have been developed that can predict the effects of amino acid mutations, as 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.

[0047] In certain embodiments, the MUC1-binding domain of the Disclosure also includes MUC1-binding domain variants that include one or more variations in the framework region in addition to the variations in the HCDR described above. In certain embodiments, the MUC1-binding domain of the present disclosure includes a heavy chain variable region having an amino acid sequence represented by any one of SEQ ID NOs: 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%, at least 85%, at least 90%, or at least 95% sequence identity to them. In certain embodiments, the MUC1-binding domain variant of the Disclosure includes one or more variations in the framework region, but does not include any variation in the CDR region. Such variants are expected to have at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity with respect to the sequences disclosed herein, and to retain MUC1-binding specificity. Therefore, in certain embodiments, the MUC1-binding domain of the Disclosure is - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with respect to the amino acid sequence shown in SEQ ID NO: 1, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 2, the HCDR2 amino acid sequence shown in SEQ ID NO: 3, and the HCDR3 amino acid sequence shown in SEQ ID NO: 4. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 5, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 6, the HCDR2 amino acid sequence shown in SEQ ID NO: 7, and the HCDR3 amino acid sequence shown in SEQ ID NO: 8. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 9, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 10, the HCDR2 amino acid sequence shown in SEQ ID NO: 11, and the HCDR3 amino acid sequence shown in SEQ ID NO: 12. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 13, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 14, the HCDR2 amino acid sequence shown in SEQ ID NO: 15, and the HCDR3 amino acid sequence shown in SEQ ID NO: 16. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 17, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 18, the HCDR2 amino acid sequence shown in SEQ ID NO: 19, and the HCDR3 amino acid sequence shown in SEQ ID NO: 20. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 21, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 22, the HCDR2 amino acid sequence shown in SEQ ID NO: 23, and the HCDR3 amino acid sequence shown in SEQ ID NO: 24. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 25, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 26, the HCDR2 amino acid sequence shown in SEQ ID NO: 27, and the HCDR3 amino acid sequence shown in SEQ ID NO: 28. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 29, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 30, the HCDR2 amino acid sequence shown in SEQ ID NO: 31, and the HCDR3 amino acid sequence shown in SEQ ID NO: 32. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 33, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 34, the HCDR2 amino acid sequence shown in SEQ ID NO: 35, and the HCDR3 amino acid sequence shown in SEQ ID NO: 36. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 37, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 38, the HCDR2 amino acid sequence shown in SEQ ID NO: 39, and the HCDR3 amino acid sequence shown in SEQ ID NO: 40. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 41, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 42, the HCDR2 amino acid sequence shown in SEQ ID NO: 43, and the HCDR3 amino acid sequence shown in SEQ ID NO: 44. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 45, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 46, the HCDR2 amino acid sequence shown in SEQ ID NO: 47, and the HCDR3 amino acid sequence shown in SEQ ID NO: 48. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 49, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 50, the HCDR2 amino acid sequence shown in SEQ ID NO: 51, and the HCDR3 amino acid sequence shown in SEQ ID NO: 52. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 53, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 54, the HCDR2 amino acid sequence shown in SEQ ID NO: 55, and the HCDR3 amino acid sequence shown in SEQ ID NO: 56. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 57, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 58, the HCDR2 amino acid sequence shown in SEQ ID NO: 60, and the HCDR3 amino acid sequence shown in SEQ ID NO: 61. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 62, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 63, the HCDR2 amino acid sequence shown in SEQ ID NO: 64, and the HCDR3 amino acid sequence shown in SEQ ID NO: 65. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 66, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 67, the HCDR2 amino acid sequence shown in SEQ ID NO: 68, and the HCDR3 amino acid sequence shown in SEQ ID NO: 69. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 70, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 71, the HCDR2 amino acid sequence shown in SEQ ID NO: 72, and the HCDR3 amino acid sequence shown in SEQ ID NO: 73. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 74, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 75, the HCDR2 amino acid sequence shown in SEQ ID NO: 76, and the HCDR3 amino acid sequence shown in SEQ ID NO: 77. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 78, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 79, the HCDR2 amino acid sequence shown in SEQ ID NO: 80, and the HCDR3 amino acid sequence shown in SEQ ID NO: 81. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with respect to the amino acid sequence shown in SEQ ID NO: 82, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 83, the HCDR2 amino acid sequence shown in SEQ ID NO: 84, and the HCDR3 amino acid sequence shown in SEQ ID NO: 85. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 86, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 87, the HCDR2 amino acid sequence shown in SEQ ID NO: 88, and the HCDR3 amino acid sequence shown in SEQ ID NO: 89. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 90, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 91, the HCDR2 amino acid sequence shown in SEQ ID NO: 92, and the HCDR3 amino acid sequence shown in SEQ ID NO: 93. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 94, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 95, the HCDR2 amino acid sequence shown in SEQ ID NO: 96, and the HCDR3 amino acid sequence shown in SEQ ID NO: 97. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 98, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 99, the HCDR2 amino acid sequence shown in SEQ ID NO: 100, and the HCDR3 amino acid sequence shown in SEQ ID NO: 101. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 102, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 103, the HCDR2 amino acid sequence shown in SEQ ID NO: 104, and the HCDR3 amino acid sequence shown in SEQ ID NO: 105. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 106, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 107, the HCDR2 amino acid sequence shown in SEQ ID NO: 108, and the HCDR3 amino acid sequence shown in SEQ ID NO: 109. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with respect to the amino acid sequence shown in SEQ ID NO: 110, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 111, the HCDR2 amino acid sequence shown in SEQ ID NO: 112, and the HCDR3 amino acid sequence shown in SEQ ID NO: 113. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 114, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 115, the HCDR2 amino acid sequence shown in SEQ ID NO: 116, and the HCDR3 amino acid sequence shown in SEQ ID NO: 117. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 118, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 119, the HCDR2 amino acid sequence shown in SEQ ID NO: 120, and the HCDR3 amino acid sequence shown in SEQ ID NO: 121. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 122, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 123, the HCDR2 amino acid sequence shown in SEQ ID NO: 124, and the HCDR3 amino acid sequence shown in SEQ ID NO: 125. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 126, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 127, the HCDR2 amino acid sequence shown in SEQ ID NO: 128, and the HCDR3 amino acid sequence shown in SEQ ID NO: 129. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 130, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 131, the HCDR2 amino acid sequence shown in SEQ ID NO: 132, and the HCDR3 amino acid sequence shown in SEQ ID NO: 133. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 134, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 135, the HCDR2 amino acid sequence shown in SEQ ID NO: 136, and the HCDR3 amino acid sequence shown in SEQ ID NO: 137. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 138, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 139, the HCDR2 amino acid sequence shown in SEQ ID NO: 140, and the HCDR3 amino acid sequence shown in SEQ ID NO: 141. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 142, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 143, the HCDR2 amino acid sequence shown in SEQ ID NO: 144, and the HCDR3 amino acid sequence shown in SEQ ID NO: 145. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 146, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 147, the HCDR2 amino acid sequence shown in SEQ ID NO: 148, and the HCDR3 amino acid sequence shown in SEQ ID NO: 149. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 150, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 151, the HCDR2 amino acid sequence shown in SEQ ID NO: 152, and the HCDR3 amino acid sequence shown in SEQ ID NO: 153. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 154, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 155, the HCDR2 amino acid sequence shown in SEQ ID NO: 156, and the HCDR3 amino acid sequence shown in SEQ ID NO: 157. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 158, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 159, the HCDR2 amino acid sequence shown in SEQ ID NO: 160, and the HCDR3 amino acid sequence shown in SEQ ID NO: 161. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 162, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 163, the HCDR2 amino acid sequence shown in SEQ ID NO: 164, and the HCDR3 amino acid sequence shown in SEQ ID NO: 165. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 166, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 167, the HCDR2 amino acid sequence shown in SEQ ID NO: 168, and the HCDR3 amino acid sequence shown in SEQ ID NO: 169. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 170, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 171, the HCDR2 amino acid sequence shown in SEQ ID NO: 172, and the HCDR3 amino acid sequence shown in SEQ ID NO: 173. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 174, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 175, the HCDR2 amino acid sequence shown in SEQ ID NO: 176, and the HCDR3 amino acid sequence shown in SEQ ID NO: 177. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 178, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 179, the HCDR2 amino acid sequence shown in SEQ ID NO: 180, and the HCDR3 amino acid sequence shown in SEQ ID NO: 181. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 182, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 183, the HCDR2 amino acid sequence shown in SEQ ID NO: 184, and the HCDR3 amino acid sequence shown in SEQ ID NO: 185. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 186, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 187, the HCDR2 amino acid sequence shown in SEQ ID NO: 188, and the HCDR3 amino acid sequence shown in SEQ ID NO: 189. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 190, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 191, the HCDR2 amino acid sequence shown in SEQ ID NO: 192, and the HCDR3 amino acid sequence shown in SEQ ID NO: 193. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 194, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 195, the HCDR2 amino acid sequence shown in SEQ ID NO: 196, and the HCDR3 amino acid sequence shown in SEQ ID NO: 197. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 198, and comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 199, the HCDR2 amino acid sequence shown in SEQ ID NO: 200, and the HCDR3 amino acid sequence shown in SEQ ID NO: 201. - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 202, and including the HCDR1 amino acid sequence shown in SEQ ID NO: 203, the HCDR2 amino acid sequence shown in SEQ ID NO: 204, and the HCDR3 amino acid sequence shown in SEQ ID NO: 205, or - A heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with the amino acid sequence shown in SEQ ID NO: 206, comprising the HCDR1 amino acid sequence shown in SEQ ID NO: 207, the HCDR2 amino acid sequence shown in SEQ ID NO: 208, and the HCDR3 amino acid sequence shown in SEQ ID NO: 209.

[0048] The MUC1-binding domains of this disclosure are generated using a common light chain, in particular a common light chain called VK1-39 / JK1. The binding domains of this disclosure include, but are not limited to, common light chains known in the art, and may include any suitable light chain. In certain embodiments, the MUC1-binding domains of this disclosure include the common light chain VK1-39 / JK1, or variants thereof having a limited number, e.g., one, two, or three non-conservative amino acid substitutions, or an unlimited number of conservative amino acid substitutions.

[0049] In certain embodiments, the MUC1-binding domain of the Disclosure comprises a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a variant thereof. In certain embodiments, the MUC1-binding domain of the Disclosure comprises a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto.

[0050] In certain embodiments, the MUC1-binding domain of the Disclosure includes a light chain variable region comprising light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), each having the amino acid sequences shown in SEQ ID NOs. 219, SEQ ID NOs. 220, and SEQ ID NOs. In certain embodiments, the light chain variable region of the MUC1-binding domain of the Disclosure also includes its variants, and each LCDR may contain up to three, two, or one amino acid substitutions. These amino acid substitutions are, for example, conservative amino acid substitutions.

[0051] In certain embodiments, the MUC1-binding domain of the Disclosure also includes MUC1-binding domain variants that include one or more variations in the framework region in addition to the variations in the LCDR described above. In certain embodiments, the MUC1-binding domain variant of the Disclosure does not include variations in the LCDR region but includes one or more variations in the framework region. Such variants have at least 80%, or at least 85%, or at least 90%, or at least 95% sequence identity with respect to the sequences disclosed herein. Therefore, in certain embodiments, the MUC1-binding domain of the Disclosure is -A light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity with respect to the amino acid sequence shown in SEQ ID NO: 218, comprising 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.

[0052] These LCDRs and / or light chain variable regions, including the light chain or light chain variable region, may be the light chain referred to in the art as VK1-39 / JK1. This is a common light chain. The term “common light chain” as used herein refers to a light chain that can pair with multiple different heavy chains, such as heavy chains having different antigen or epitope binding specificities. The term “common light chain” encompasses light chains that are identical or have some amino acid sequence differences, while the binding specificity of the full-length antibody is not affected. For example, it is possible, within the scope of the definition of a common light chain as used herein, to prepare or find light chains that are not identical but are still functionally equivalent by using well-established variations that introduce conservative amino acid changes, changes in amino acids in regions known to not contribute to or only partially contribute to binding specificity when paired with a heavy chain.

[0053] Apart from the common light chains including the LCDRs and / or light chain variable regions described above, other common light chains known in the art may be used. Examples of such common light chains include, but are not limited to, VK1-39 / JK5, ​​which includes a light chain variable region comprising light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3) having the amino acid sequence shown in SEQ ID NO: 229. In certain embodiments, the light chain includes a light chain variable region comprising light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3) having the amino acid sequence shown in SEQ ID NO: 229, where each LCDR may contain up to three, two, or one amino acid variation, e.g., substitutions. In certain embodiments, the light chain includes a light chain variable region having the amino acid sequence shown in SEQ ID NO: 229, 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 light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), each having the amino acid sequences shown in SEQ ID NOs. 230, 231, and 232, respectively, and VK3-15 / JK1 comprises light chain variable regions having the amino acid sequence shown in SEQ ID NOs. In certain embodiments, the light chain comprises light chain variable regions including light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), each of which may contain up to three, two, or one amino acid variation, e.g., substitutions. In certain embodiments, the light chain includes a light chain variable region having the amino acid sequence shown in SEQ ID NO: 234, 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 light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), each having the amino acid sequences shown in SEQ ID NO: 235, SEQ ID NO: 236, and SEQ ID NO: 237, respectively, and VK3-20 / JK1 comprises light chain variable regions having the amino acid sequence shown in SEQ ID NO: 239, comprising light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3). In certain embodiments, the light chain comprises light chain variable regions comprising light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), each of which may contain up to three, two, or one amino acid variation, e.g., substitutions. In certain embodiments, the light chain includes a light chain variable region having the amino acid sequence shown in SEQ ID NO: 239, or having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, the light chain includes light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), each having the amino acid sequences shown in SEQ ID NO: 240, SEQ ID NO: 241, and SEQ ID NO: 242, respectively, and VL3-21 / JL3 includes light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), which are light chain variable regions having the amino acid sequence shown in SEQ ID NO: 244. In certain embodiments, the light chain comprises a light chain variable region having the amino acid sequence shown in SEQ ID NO: 244, including light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), where each LCDR may contain up to three, two, or one amino acid variation, such as a substitution. In certain embodiments, the light chain comprises a light chain variable region having the amino acid sequence 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 a particular embodiment, the light chain comprises light chain CDR1(LCDR1), light chain CDR2(LCDR2), and light chain CDR3(LCDR3), each having the amino acid sequences shown in SEQ ID NO: 245, SEQ ID NO: 246, and SEQ ID NO: 247, respectively.

[0054] VK1-39 is an abbreviation for the immunoglobulin variable kappa 1-39 gene. This gene is also known as immunoglobulin variable kappa 1-39, IGKV139, IGKV1-39, and IgVκ1-39. The external ID for this gene is HGNC:5740;Entrez Gene:28930;Ensembl:ENSG00000242371. The amino acid sequence for VK1-39 is given as SEQ ID NO: 227. This is the sequence of the V region. The V region can be combined with one of the five J regions. Preferred VJ region sequences are indicated as VK1-39 / JK1 (SEQ ID NO: 228) and VK1-39 / JK5 (SEQ ID NO: 229), with alternative names being IgVκ1-39*01 / IGJκ1*01 or IgVκ1-39*01 / IGJκ5*01 (named by the IMGT database at imgt.org). These names are illustrative and encompass allele variants of the gene segment.

[0055] VK3-15 is an abbreviation for the immunoglobulin variable kappa 3-15 gene. This gene is also known as immunoglobulin variable kappa 3-15, IGKV315, IGKV3-15, and IgVκ3-15. The external ID for this gene is HGNC:5816;Entrez Gene:28913;Ensembl:ENSG00000244437. The amino acid sequence for VK3-15 is given as SEQ ID NO: 233. This is the sequence of the V region. The V region can be combined with one of five J regions. A preferred VJ region sequence is indicated as VK3-15 / JK1 (SEQ ID NO: 234), with the alternative name Vκ3-15*01 / IGJκ1*01 (named by the IMGT database World Wide Web on imgt.org). This name is illustrative and encompasses allele variants of the gene segment.

[0056] VK3-20 is an abbreviation for the immunoglobulin variable kappa 3-20 gene. This gene is also known as immunoglobulin variable kappa 3-20, IGKV320, IGKV3-20, and IgVκ3-20. The external ID for this gene is HGNC:5817;Entrez Gene:28912;Ensembl:ENSG00000239951. The amino acid sequence for VK3-20 is like SEQ ID NO: 238. This is the sequence of the V region. The V region can be combined with one of five J regions. A preferred VJ region sequence is indicated as VK3-20 / JK1 (SEQ ID NO: 239), with the alternative name IgVκ3-20*01 / IGJκ1*01 (named by the IMGT database World Wide Web on imgt.org). This name is illustrative and encompasses allele variants of the gene segment.

[0057] VL3-21 is an abbreviation for the immunoglobulin variable lambda 3-21 gene. This gene is also known as immunoglobulin variable lambda 3-21, IGLV321, IGLV3-21, and IgVλ3-21. The external ID for this gene is HGNC:5905;Entrez Gene:28796;Ensembl:ENSG00000211662.2. The amino acid sequence for VL3-21 is given as SEQ ID NO: 243. This is the sequence of the V region. The V region can be combined with one of five J regions. A preferred VJ region sequence is indicated as VL3-21 / JL3 (SEQ ID NO: 244), with the alternative name IgVλ3-21 / IGJλ3 (named by the IMGT database World Wide Web on imgt.org). This name is illustrative and encompasses allele variants of the gene segment.

[0058] Furthermore, any light chain variable region of a MUC1 antibody available in the art, or any other light chain variable region readily obtainable from an antibody display library by exhibiting antigen-binding activity when paired with the MUC1-binding domain of the present disclosure, may be used.

[0059] In certain embodiments, the MUC1-binding domain of the binding portion of the present disclosure may further include CH1 and CL regions. Any CH1 domain, specifically a human CH1 domain, may be used. An example of a preferred CH1 domain is provided by the amino acid sequence provided as SEQ ID NO: 223. Any CL domain, specifically a human CL domain, may be used. An example of a preferred CL domain is provided by the amino acid sequence provided as SEQ ID NO: 226.

[0060] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0061] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 6, SEQ ID NO: 7, and SEQ ID NO: 8, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0062] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 10, SEQ ID NO: 11, and SEQ ID NO: 12, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0063] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 16, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0064] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0065] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO: 24, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0066] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 28, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0067] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 30, SEQ ID NO: 31, and SEQ ID NO: 32, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0068] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0069] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO: 40, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0070] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 42, SEQ ID NO: 43, and SEQ ID NO: 44, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0071] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 46, SEQ ID NO: 47, and SEQ ID NO: 48, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0072] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO: 52, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0073] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 54, SEQ ID NO: 55, and SEQ ID NO: 56, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0074] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 58, SEQ ID NO: 60, and SEQ ID NO: 61, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0075] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO: 65, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0076] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 67, SEQ ID NO: 68, and SEQ ID NO: 69, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0077] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 71, SEQ ID NO: 72, and SEQ ID NO: 73, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0078] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 75, SEQ ID NO: 76, and SEQ ID NO: 77, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0079] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 79, SEQ ID NO: 80, and SEQ ID NO: 81, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0080] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 83, SEQ ID NO: 84, and SEQ ID NO: 85, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0081] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 87, SEQ ID NO: 88, and SEQ ID NO: 89, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0082] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 91, SEQ ID NO: 92, and SEQ ID NO: 93, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0083] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 95, SEQ ID NO: 96, and SEQ ID NO: 97, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0084] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 99, SEQ ID NO: 100, and SEQ ID NO: 101, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0085] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 103, SEQ ID NO: 104, and SEQ ID NO: 105, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0086] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 107, SEQ ID NO: 108, and SEQ ID NO: 109, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0087] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 111, SEQ ID NO: 112, and SEQ ID NO: 113, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0088] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 115, SEQ ID NO: 116, and SEQ ID NO: 117, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0089] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 119, SEQ ID NO: 120, and SEQ ID NO: 121, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0090] In one embodiment, this disclosure, - Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 123, SEQ ID NO: 124, and SEQ ID NO: 125, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0091] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO: 129, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0092] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 131, SEQ ID NO: 132, and SEQ ID NO: 133, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0093] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 135, SEQ ID NO: 136, and SEQ ID NO: 137, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0094] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140, and SEQ ID NO: 141, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0095] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 143, SEQ ID NO: 144, and SEQ ID NO: 145, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0096] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 147, SEQ ID NO: 148, and SEQ ID NO: 149, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0097] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 151, SEQ ID NO: 152, and SEQ ID NO: 153, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0098] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 155, SEQ ID NO: 156, and SEQ ID NO: 157, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0099] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 159, SEQ ID NO: 160, and SEQ ID NO: 161, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0100] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 163, SEQ ID NO: 164, and SEQ ID NO: 165, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0101] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 167, SEQ ID NO: 168, and SEQ ID NO: 169, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0102] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 171, SEQ ID NO: 172, and SEQ ID NO: 173, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0103] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 175, SEQ ID NO: 176, and SEQ ID NO: 177, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0104] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 179, SEQ ID NO: 180, and SEQ ID NO: 181, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0105] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 183, SEQ ID NO: 184, and SEQ ID NO: 185, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0106] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188, and SEQ ID NO: 189, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0107] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 191, SEQ ID NO: 192, and SEQ ID NO: 193, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0108] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 195, SEQ ID NO: 196, and SEQ ID NO: 197, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0109] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200, and SEQ ID NO: 201, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0110] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 203, SEQ ID NO: 204, and SEQ ID NO: 205, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0111] In one embodiment, this disclosure, -Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 207, SEQ ID NO: 208, and SEQ ID NO: 209, The present invention provides an MUC1-binding domain comprising 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, and a light chain CDR3 (LCDR3) having the amino acid sequence shown in SEQ ID NO: 221. Each of the HCDR and / or LCDR may contain up to three, two, or one amino acid variation, such as substitution. In certain embodiments, the HCDR and / or LCDR do not contain any amino acid variations.

[0112] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 1, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0113] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 5, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0114] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 9, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0115] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 13, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0116] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 17, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0117] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 21, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0118] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 25, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0119] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 29, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0120] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 33, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0121] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 37, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0122] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 41, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0123] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 45, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0124] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 49, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0125] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 53, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0126] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 57, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0127] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 62, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0128] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 66, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0129] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 70, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0130] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 74, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0131] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 78, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0132] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 82, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0133] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 86, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0134] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 90, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0135] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 94, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0136] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 98, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0137] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 102, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0138] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 106, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0139] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 110, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0140] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 114, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0141] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 118, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0142] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 122, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0143] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 126, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0144] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 130, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0145] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 134, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0146] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 138, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0147] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 142, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0148] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 146, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0149] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 150, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0150] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 154, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0151] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 158, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0152] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 162, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0153] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 166, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0154] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 170, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0155] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 174, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0156] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 178, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0157] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 182, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0158] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 186, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0159] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 190, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0160] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 194, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0161] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 198, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - Provides a MUC1 binding domain comprising a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, or a light chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises an HCDR and an LCDR that do not contain amino acid variations, respectively. In certain embodiments, each of the heavy chain variable region and the light chain variable region does not contain amino acid variations.

[0162] In one embodiment, this disclosure, - A heavy chain variable region having the amino acid sequence shown in Sequence ID No. 202, or a heavy chain variable region having at least 80%, at least 85%, at least 90%, or at least 95% sequence identity thereto, - A light chain variable region having the amino acid sequence 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, and a MUC1 binding domain comprising the same are provided. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises HCDRs and LCDRs without amino acid variations. In certain embodiments, each of the heavy chain variable region and the light chain variable region has no amino acid variations.

[0163] In one embodiment, the present disclosure - A heavy chain variable region having the amino acid sequence 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 - A light chain variable region having the amino acid sequence 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, and a MUC1 binding domain comprising the same are provided. In certain embodiments, each of the heavy chain variable region and the light chain variable region comprises HCDRs and LCDRs without amino acid variations. In certain embodiments, each of the heavy chain variable region and the light chain variable region has no amino acid variations.

[0164] "Percent identity (%)" with respect to a nucleic acid sequence or an amino acid sequence 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. Gaps may be introduced into either of the two sequences being compared to optimize the alignment between these two sequences. Such alignment can be performed over the entire length of the sequences being compared. Alternatively, the alignment can be performed over a shorter length, e.g., over about 20, about 50, about 100 or more nucleic acids / bases or amino acids. Sequence identity is the percentage of exact matches between these two sequences over the reported aligned regions.

[0165] The comparison of arrays and the determination of the percentage of array identity between two arrays can be achieved using mathematical algorithms. A person skilled in the art will recognize the fact that several different computer programs are available for aligning two arrays and determining the identity between the two arrays (Kruskal JB. SIAM Review. 1983;25(2),201-237). The percentage of sequence identity between two amino acid sequences or nucleic acid sequences can be determined using the Needleman-Wunsch algorithm for alignment of the two sequences. (Needleman SB, Wunsch CD. J Mol Biol. 1970;48(3):443-53). The Needleman-Wunsch algorithm is implemented in the computer program NEEDLE. For the purposes of the present invention, the NEEDLE program from the EMBOSS package 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 as the substitution matrix. For DNA sequences, DNAFULL is used. The parameters used are a gap open penalty of 10 and a gap extension penalty of 0.5.

[0166] After alignment by the program NEEDLE as described above, the percentage of sequence identity between the query sequence and the sequence of the present invention is calculated as follows: The number of corresponding positions in the alignment represents the number of identical amino acids or identical nucleotides in both sequences, divided by the total length of the alignment after subtracting the total number of gaps in the alignment.

[0167] In certain embodiments, the disclosure also provides a binding portion including the MUC1 binding domains described herein. In certain embodiments, the binding portion of the disclosure includes two MUC1 binding domains described herein. In certain embodiments, the binding portion of the disclosure consists of two MUC1 binding domains and an Fc region. In certain embodiments, the Fc region includes hinge, CH2, and CH3 domains.

[0168] The "binding region" refers to a proteinaceous molecule containing a binding domain. Examples include full-length IgG antibodies, immune complexes, diabodies, BiTEs, Fab fragments, scFvs, tandem scFvs, and single-domain antibodies (V). HH and V H This includes all antibody formats available in the art, such as minibodies, scFab, scFv-zippers, nanobodies, DART molecules, TandAb, Fab-scFv, F(ab)'2, F(ab)'2-scFv2, and intrabodies.

[0169] In certain embodiments, the binding portion of the Disclosure is a monospecific binding portion, in particular a monospecific antibody. A monospecific antibody according to the Disclosure is an antibody comprising one or more binding domains having specificity for a single target in any antibody format. In certain embodiments, the monospecific binding portion of the Disclosure may further comprise an Fc region or a portion thereof. In certain embodiments, the monospecific binding portion of the Disclosure is an IgG1 antibody.

[0170] The Fc region mediates antibody effector functions such as complement-dependent cell-mediated cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC), and antibody-dependent phagocytosis (ADCP). Depending on the application of therapeutic antibodies or Fc fusion proteins, it may be desirable to reduce or increase effector function.

[0171] In certain embodiments, the binding portion of the Disclosure has Fc effector function. In certain embodiments, the binding portion of the Disclosure has enhanced Fc effector function. In certain embodiments, the binding portion of the Disclosure exhibits antibody-dependent cell-mediated cytotoxicity (ADCC). Binding portions, such as antibodies, can be manipulated to enhance ADCC activity (see Kubota T et al. Cancer Sci. 2009;100(9):1566-72 for a review). For example, if an 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). Modifications may also be made to improve storage or production, or to remove C-terminal lysine (Kubota T et al. Cancer Sci. 2009;100(9):1566-72). Another method to improve the ADCC activity of an antibody is by enzymatically interfering with the glycosylation pathway that results in reduced fucose (von Horsten HH. et al. Glycobiology. 2010;20(12):1607-18). Alternatively or additionally, several other strategies can be used to achieve enhancement of ADCC, including, for example, glycotechnology (Kyowa Hakko / Biowa, GlycArt (Roche) and Eureka Therapeutics) and mutagenesis, all of which aim to improve Fc binding to low-affinity activated FcγRIIIa and / or reduce binding to low-affinity inhibitory FcγRIIb. In certain embodiments, the binding moiety of this disclosure exhibits enhanced antibody-dependent cell-mediated cytotoxicity (ADCC). In certain embodiments, the binding moiety of this disclosure is afucosylated.

[0172] In certain embodiments, the binding portion of this disclosure has a higher binding signal than the reference antibody. In certain embodiments, the binding signal is such that it is measured by a FACS assay using MUC1-Tn expressing cells or MUC-1-STn expressing cells. 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 generated 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 generated by stably transfecting T47D cells with a plasmid encoding ST6GALNAC, as described herein. In a particular embodiment, the reference antibody is a bivalent monospecific antibody comprising two heavy chains having the amino acid sequence shown in SEQ ID NO: 210 and two light chains having the amino acid sequence shown in SEQ ID NO: 211.

[0173] In certain embodiments, the Disclosure provides a binding portion comprising the MUC1 binding domain of the Disclosure, -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 1, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 5, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 9, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 13, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 17, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 21, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 25, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 29, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 33, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 37, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 41, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 45, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 49, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 53, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 57, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 62, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 66, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 70, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 74, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 78, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 82, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 86, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 90, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 94, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 98, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 102, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 106, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 110, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 114, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 118 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 122 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 126 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 130 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 134 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 138 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 142 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 146 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 150 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 154 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 158 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, - An antibody comprising two heavy-chain variable regions having the amino acid sequence shown in SEQ ID NO: 162 and a light-chain variable region having the amino acid sequence shown in SEQ ID NO: 218, -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 166, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 170, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 174, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 178, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 182, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 186, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 190, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 194, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 198, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 202, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, and -An antibody comprising two heavy chain variable regions having the amino acid sequence shown in SEQ ID NO: 206, and a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218. Alternatively, the following group is selected: a binding site that competes with the antibody for binding to the MUC1 peptide containing the amino acid sequence PAPGSTAPPAHGVT*SAPDTRPAPG (SEQ ID NO: 249) and / or PAPGSTAPPAHGVT*S*APDTRPAPG (SEQ ID NO: 249), where * represents Tn or STn glycosylation.

[0174] This specification further provides nucleic acids useful for producing the MUC1 binding domain of the Disclosure, or binding moieties containing the MUC1 binding domain. In certain embodiments, such nucleic acids include a nucleic acid sequence encoding the heavy chain variable region of the MUC1 binding domain described herein. In certain embodiments, the nucleic acids of the Disclosure may further include a CH1 region and, optionally, a nucleic acid sequence encoding the hinge, CH2, and / or CH3 regions. In certain embodiments, the nucleic acids of the Disclosure may further include a light chain variable region and, optionally, at least one nucleic acid sequence encoding the CL region. In certain embodiments, the light chain variable region may be the common light chain variable region described herein.

[0175] This specification further provides vectors useful for producing the MUC1 binding domain of this disclosure, or binding moieties containing the MUC1 binding domain. In certain embodiments, such expression vectors include a nucleic acid sequence encoding the heavy chain variable region of the MUC1 binding domain described herein. In certain embodiments, the vectors of this disclosure may further include a CH1 region, and optionally a nucleic acid sequence encoding the hinge, CH2, and / or CH3 regions. In certain embodiments, the vectors of this disclosure may further include a light chain variable region, and optionally at least one nucleic acid sequence encoding the CL region. In certain embodiments, the light chain variable region may be the common light chain variable region described herein.

[0176] In certain embodiments, the Disclosure also provides cells comprising nucleic acid sequences encoding the heavy chain variable region of the MUC1 binding domain described herein. In certain embodiments, the cells of the Disclosure may further comprise the CH1 region and, optionally, the hinge, CH2, and / or CH3 regions. In certain embodiments, the cells of the Disclosure may further comprise the light chain variable region and, optionally, at least one nucleic acid sequence encoding the CL region. In certain embodiments, the light chain variable region may be the common light chain variable region described herein.

[0177] In certain embodiments, the Disclosure also provides cells that produce the MUC1 binding domain described herein, or binding moieties comprising the MUC1 binding domain. In certain embodiments, such cells may be recombinant cells transformed with the vector of the Disclosure. In certain embodiments, the cells of the Disclosure include a nucleic acid sequence encoding the heavy chain variable region of the MUC1 binding domain described herein. In certain embodiments, the cells of the Disclosure further include a CH1 region, and optionally a nucleic acid sequence encoding the hinge, CH2, and / or CH3 regions. In certain embodiments, the cells of the Disclosure further include a light chain variable region, in particular the light chain variable region described herein, and optionally at least one nucleic acid sequence encoding the CL region.

[0178] In certain embodiments, the Disclosure provides a pharmaceutical composition comprising an effective amount of a MUC1-binding domain or a binding moiety containing a MUC1-binding domain as described herein, and optionally a pharmaceutically acceptable carrier.

[0179] In certain embodiments, this disclosure provides a MUC1-binding domain as described herein, or a binding moiety comprising a MUC1-binding domain as described herein, and a pharmaceutical composition as described herein, for use in therapy.

[0180] In certain embodiments, this disclosure provides a MUC1-binding domain described herein, or a binding moiety comprising a MUC1-binding domain, or a pharmaceutical composition described herein, for use in the treatment of cancer.

[0181] In certain embodiments, the present disclosure provides a method for treating a disease, comprising administering to an individual in need of treatment an effective amount of the MUC1-binding domain described herein, or a binding moiety comprising the MUC1-binding domain, or a pharmaceutical composition described herein.

[0182] In certain embodiments, the present disclosure provides a method for treating cancer, comprising administering to an individual in need of treatment an effective amount of the MUC1-binding domain described herein, or a binding moiety containing the MUC1-binding domain, or a pharmaceutical composition described herein.

[0183] As used herein, the terms “individual,” “subject,” and “patient” are interchangeable and refer to mammals such as humans, mice, rats, hamsters, guinea pigs, rabbits, cats, dogs, monkeys, cattle, horses, and pigs, in particular humans with cancer.

[0184] As used herein, the terms “to treat,” “to treat,” and “treatment” refer to any type of intervention or process performed on or involving the administration of an active agent or combination of active agents to a subject for the purpose of curing or improving a disease or condition of the subject, or for the purpose of producing a positive therapeutic response. As used herein, “positive therapeutic response” refers to a beneficial effect, such as reversing, alleviating, improving, suppressing, or delaying a disease-related symptom, complication, condition, or biochemical sign, and preventing the onset, progression, development, worsening, or recurrence of a disease-related symptom, complication, condition, or biochemical sign, such as a treatment that produces improvement in at least one symptom of a disease or disorder, such as cancer. A beneficial effect can take the form of improvement above baseline, including improvement beyond measurements or observations made before initiating treatment according to the method. For example, a beneficial effect can take the form of delaying, stabilizing, stopping, or reversing the progression of cancer in a subject at any clinical stage, as demonstrated by a reduction or elimination of the clinical or diagnostic symptoms of the disease, or a marker of cancer. Effective treatments may, for example, reduce tumor size, decrease the presence of circulating tumor cells, mitigate or prevent tumor metastasis, delay or halt tumor growth, and / or prevent or delay tumor recurrence or recurrence.

[0185] The terms "therapeutic dose" or "effective dose" refer to the amount of a drug or combination of drugs used to treat a disease such as cancer. In some embodiments, the therapeutic dose is an amount sufficient to delay tumor development. In some embodiments, the therapeutic dose is an amount sufficient to prevent or delay tumor recurrence.

[0186] As used herein, an effective amount of a drug or composition can, for example, (i) reduce the number of cancer cells, (ii) reduce tumor size, (iii) inhibit, delay, slow to some extent, and stop the invasion of cancer cells into peripheral organs, (iv) inhibit tumor metastasis, (v) inhibit tumor growth, (vi) prevent or delay tumor development and / or recurrence, and / or (vii) alleviate to some extent one or more of the symptoms associated with cancer.

[0187] The effective dose may vary depending on factors such as the disease state, age, sex, and weight of the individual being treated, as well as the ability of the drug or drug combination to induce the desired response in the individual, which can be easily assessed by a regular, experienced physician or other healthcare worker.

[0188] An effective dose can be administered to the subject in one or more doses.

[0189] The effective dose may also include the amount that balances any toxic or harmful effects of the drug or combination of drugs with the beneficial effects.

[0190] The term "pharmaceutical" refers to a therapeutically active substance, in this case, the binding domain or binding portion of the Disclosure, or the pharmaceutical composition of the Disclosure.

[0191] As used herein, “includes” and its conjugations are used in their non-restrictive sense, meaning that the item preceding the word is included, but items not specifically mentioned are not excluded.

[0192] The articles "a" and "an" are used herein to refer to one or more of the grammatical objects of the article. For example, "element" means one or more elements.

[0193] No reference in this specification to patent documents or other matters shall be deemed to be an admission that such documents or matters were known, or that the information contained herein was part of the common general knowledge as of the priority date of any of the claims.

[0194] All patent and document references cited herein are incorporated herein by reference in their entirety.

[0195] In this specification, unless otherwise specified, amino acid positions assigned to the CDR and framework within the variable region of an antibody or antibody fragment are designated by Kabat numbering (see Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md., 1987 and 1991)). Amino acids within the constant region are indicated by the EU numbering system.

[0196] Acceptance numbers are primarily provided to offer a method for further target identification, and the actual sequence of the bound protein may change due to mutations in the coding gene, such as mutations that occur in some cancers. The antigen-binding sites of the binding domain or binding portion of this disclosure can bind to antigens and their various variants, for example, those expressed by some antigen-positive immune cells or tumor cells. HGNC is an abbreviation for the Human Gene Nomenclature Committee (HUGO). The number following the abbreviation is an acceptance number that allows information about the gene and the protein encoded by that gene to be retrieved from the HGNC database. Entrez Gene provides an acceptance number or gene ID that allows information about the gene or the protein encoded by that gene to be retrieved from the NCBI (National Center for Biotechnology Information) database. Ensembl provides an acceptance number that allows information about the gene or the protein encoded by that gene to be obtained from the Ensemble database. Ensembl is a joint project between EMBL-EBI and the Wellcome Trust Sanger Institute to develop a software system that generates and maintains automated annotations for selected eukaryotic genomes.

[0197] Where a gene or protein is referred to herein, preferably the human form of that gene or protein is referred to. Where a gene or protein is referred to herein, both native genes or proteins and variants of that gene or protein that can be detected in tumors, cancers, etc., or human tumors, cancers, etc., are referred to. [Brief explanation of the drawing]

[0198] The following nomenclature rules are used herein as follows: In the figures, the reference bivalent monospecific antibody is shown in the format of Sequence ID A / B, where Sequence ID A refers to the heavy chain of both binding domains and Sequence ID B refers to the light chain of both binding domains. The bivalent monospecific MUC1 IgG of this disclosure as illustrated herein includes the light chain of Sequence ID 217 and the CH1, hinge, CH2, and CH3 regions of Sequence IDs 223, 222, 224, and 225, respectively, and is shown in the format of Sequence ID A, referring to the heavy chain variable region of both binding domains.

[0199] [Figure 1] This is a schematic diagram of the MUC1 structure. A) is the structure of normal MUC1, and B) is the structure of abnormal MUC1 (extracted from Figure 1 in Gao T. et al. Biomed Pharmacother. 2020). Cancer-associated (CA) MUC1 epitopes include the exposed core peptide, MUC1-Tn, and MUC1-sTn. [Figure 2-1] FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-2] FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-3] FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-4] FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-5]FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-6] FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-7] FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 2-8]FACS data (Graphs A, C, E, G, I, K, M, and O) showing the specificity and relative affinity of MUC1 IgG to MCF7 cells compared to MCF10A cells (Graphs B, D, F, H, J, L, N, and P). MUC1 IgG includes a core conjugate (CB), a Tn conjugate (TnB), and a (S)Tn conjugate (SEQ ID NO). Negative control antibodies are SEQ ID NOs. 216 / 217, and positive control antibodies are SEQ ID NOs. 210 / 211 (AP) and 212 / 213 (OP). X-axis: IgG concentration (μg / ml), Y-axis: median PE signal × 10⁴. [Figure 3] This graph plots the affinity of MUC1-(S)Tn for MUC1-STn against the affinity of the MUC1-(S)Tn conjugate. X-axis: IgG affinity for MUC1-STn, measured by ELISA and expressed at AUC normalized to reference antibody 5E5 and isotype control analogs. Y-axis: Fab affinity for MUC1-Tn, measured by ELISA and expressed at AUC normalized to reference antibody 5E5 and isotype control analogs. The squares within the circles represent two very similar HCDR1-based antibody groups: 1) SEQ ID NO: 1, SEQ ID NO: 29, SEQ ID NO: 45, SEQ ID NO: 57, SEQ ID NO: 78; 2) SEQ ID NO: 5, SEQ ID NO: 9, SEQ ID NO: 178. [Figure 4-1] This is glycopeptide array data providing EC50 values ​​(ug / ml) of MUC1 IgG binding to MUC1-Tn glycopeptides T1-T12 and S1-S12. No values ​​were found indicating that binding was not detected. [Figure 4-2] This is glycopeptide array data providing EC50 values ​​(ug / ml) of MUC1 IgG binding to MUC1-Tn glycopeptides T1-T12 and S1-S12. No values ​​were found indicating that binding was not detected. [Figure 5-1]These are FACS data showing the affinity of MUC1 IgG to T47D huGalNAc cells expressing MUC1-STn (Graphs A-D) and T47D cells expressing MUC1-Tn (Graphs E-H). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AH) and SEQ ID NO: 212 / 213 (D;H). The X axis represents the concentration of IgG (μg / ml), and the Y axis represents the median fluorescence value × 10⁴. [Figure 5-2] These are FACS data showing the affinity of MUC1 IgG to T47D huGalNAc cells expressing MUC1-STn (Graphs A-D) and T47D cells expressing MUC1-Tn (Graphs E-H). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AH) and SEQ ID NO: 212 / 213 (D;H). The X axis represents the concentration of IgG (μg / ml), and the Y axis represents the median fluorescence value × 10⁴. [Figure 5-3] These are FACS data showing the affinity of MUC1 IgG to T47D huGalNAc cells expressing MUC1-STn (Graphs A-D) and T47D cells expressing MUC1-Tn (Graphs E-H). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AH) and SEQ ID NO: 212 / 213 (D;H). The X axis represents the concentration of IgG (μg / ml), and the Y axis represents the median fluorescence value × 10⁴. [Figure 5-4] These are FACS data showing the affinity of MUC1 IgG to T47D huGalNAc cells expressing MUC1-STn (Graphs A-D) and T47D cells expressing MUC1-Tn (Graphs E-H). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AH) and SEQ ID NO: 212 / 213 (D;H). The X axis represents the concentration of IgG (μg / ml), and the Y axis represents the median fluorescence value × 10⁴. [Figure 6-1]These are FACS data showing the affinity of MUC1 IgG for MC38 huMUC1 COSMC KO cells (Graphs A, C, D, F, G, I, and J) compared to MC38 huMUC1 cells (Graphs B, E, H, and K). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). The X-axis represents IgG concentration (μg / ml), and the Y-axis represents the median fluorescence value × 10⁴. [Figure 6-2] These are FACS data showing the affinity of MUC1 IgG for MC38 huMUC1 COSMC KO cells (Graphs A, C, D, F, G, I, and J) compared to MC38 huMUC1 cells (Graphs B, E, H, and K). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). The X-axis represents IgG concentration (μg / ml), and the Y-axis represents the median fluorescence value × 10⁴. [Figure 6-3] These are FACS data showing the affinity of MUC1 IgG for MC38 huMUC1 COSMC KO cells (Graphs A, C, D, F, G, I, and J) compared to MC38 huMUC1 cells (Graphs B, E, H, and K). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). The X-axis represents IgG concentration (μg / ml), and the Y-axis represents the median fluorescence value × 10⁴. [Figure 6-4] These are FACS data showing the affinity of MUC1 IgG for MC38 huMUC1 COSMC KO cells (Graphs A, C, D, F, G, I, and J) compared to MC38 huMUC1 cells (Graphs B, E, H, and K). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). The X-axis represents IgG concentration (μg / ml), and the Y-axis represents the median fluorescence value × 10⁴. [Figure 6-5]These are FACS data showing the affinity of MUC1 IgG for MC38 huMUC1 COSMC KO cells (Graphs A, C, D, F, G, I, and J) compared to MC38 huMUC1 cells (Graphs B, E, H, and K). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). The X-axis represents IgG concentration (μg / ml), and the Y-axis represents the median fluorescence value × 10⁴. [Figure 6-6] These are FACS data showing the affinity of MUC1 IgG for MC38 huMUC1 COSMC KO cells (Graphs A, C, D, F, G, I, and J) compared to MC38 huMUC1 cells (Graphs B, E, H, and K). The negative control antibody is SEQ ID NO: 216 / 217, and the positive control antibodies are SEQ ID NO: 210 / 211 (AK) and SEQ ID NO: 212 / 213 (JK). The X-axis represents IgG concentration (μg / ml), and the Y-axis represents the median fluorescence value × 10⁴. [Examples]

[0200] In the examples used to illustrate the disclosure, but not intended to limit the disclosure in any way, the MUC1 binding domains of the disclosure are characterized in IgG format, and each MUC1 binding domain includes a light chain variable region having the amino acid sequence shown in SEQ ID NO: 218, a light chain constant region having the amino acid sequence shown in SEQ ID NO: 226, and a Fab format. In the examples used to illustrate the disclosure, but not intended to limit the disclosure in any way unless otherwise specified, MUC1 IgG was screened in IgG1 format, including CH1 having the amino acid sequence shown in SEQ ID NO: 223, CH2 having the amino acid sequence shown in SEQ ID NO: 224, and CH3 having the amino acid sequence shown in SEQ ID NO: 225.

[0201] The reference antibody and control antibody used in the examples are: - This is the reference antibody 5E5, an analog of the bivalent monospecific humanized antibody 5E5, and contains two heavy chains having the amino acid sequence shown in SEQ ID NO: 210 and two light chains having the amino acid sequence shown in SEQ ID NO: 211. Humanized 5E5 binds to the MUC1-(s)Tn(GS*T*A) epitope.

[0202] - The reference antibody is pankomab, an analog of the bivalent monospecific antibody pankomab, and comprises two heavy chains having the amino acid sequence shown in SEQ ID NO: 212 and two light chains having the amino acid sequence shown in SEQ ID NO: 213. Pankomab binds to the MUC1-(s)Tn(PDT*RP) epitope.

[0203] - The reference antibody is HT186-D11, an analog of the bivalent monospecific antibody HT186-D11, and contains two heavy chains having the amino acid sequence shown in SEQ ID NO: 214 and two light chains having the amino acid sequence shown in SEQ ID NO: 215. HT186-D11 binds to the MUC1 core (RPAP) epitope.

[0204] -STn-binding, bivalent, monospecific control anti-STn antibody clone 3F1 (SBH Sciences, catalog number SBH-ASTn).

[0205] -A bivalent, monospecific anti-Tn antibody clone 5F4 (SBH Sciences, catalog number SBH-Tn) that binds to Tn.

[0206] - A bivalent monospecific control antibody, VU4H5 (BioKE, catalog number 4538S), that binds to the MUC1 core (APDTRPAP) epitope.

[0207] A divalent 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.

[0208] Example 1 - Generation of MUC1 Fab Antibodies comprising a binding domain and a heavy chain variable region that binds cancer-related MUC-1 were obtained by immunizing transgenic mice (MeMo® mice) comprising a common IGKV1-39 light chain. Several immunization methods were used, including immunization with a KHL conjugate of human MUC1 40 Tn8 or MUC1 40 Tn 10 peptide, and additionally immunization with cells overexpressing human MUC1-(S)Tn. A phage display library was generated and screened in ELISA and FACS using MUC1 peptides and cell lines to identify Fabs that bind MUC1-Tn and / or MUC1-STn (MUC1-(S)Tn). Subsequently, for further characterization, the binders were recloned into an IgG format. The binding domain sequences of the present specification, once characterized and sequenced by the techniques provided herein, can then be obtained by any method known in the art.

[0209] Example 2 - Specificity and relative affinity of MUC1-(S)Tn measured using ELISA A panel of MUC1 IgGs was screened by ELISA to characterize their binding specificity and relative affinity for different MUC1 peptides. Only mock peptides and secondary antibody were included as negative assay controls. Isotype control antibodies and an analog of the reference antibody 5E5 were included as negative and positive antibody controls, respectively.

[0210] Biotinylated MUC1 and control peptides were captured at the concentrations shown in Table 1 on Maxisorp plates coated with neutravidin (ThermoFisher Scientific, catalog number 31000) and incubated at room temperature for 1 hour. MUC1 IgG and control antibody were diluted in blocking buffer and added at 10, 1, 0.1, and 0.01 ug / ml, and incubated at room temperature for 1 hour. Subsequently, the samples were incubated with diluted HRP-conjugated secondary antibody (Bethyl Laboratories, A80-104P, 1:2000) for 1 hour at room temperature. After each incubation step, three washing steps were performed using PBST. The reaction was stopped using 1 M H2SO4, and the signal was developed using TMB peroxidase substrate solution (BD Biosciences, catalog number 555214). The plates were then scanned using a microplate reader. 450 This was analyzed. [Table 1]

[0211] The assay is MUC1 40 Tn 10 Many antibodies within the MUC1 IgG panel were identified that bound to the core peptide but not to the mock peptide (data not shown). The binding specificity of MUC1-(S)Tn antibodies was classified into three groups: 1) MUC1 core peptide binder (MUC1 40 and MUC1 40 Tn 10 and MUC1 40 STn 10 1) MUC1-Tn binder (MUC1 40 Tn 10 It binds to MUC1 40 STn 10 3) MUC1-(S)Tn binder (MUC1 40 Tn 10 and MUC1 40 STn 10 Binning was performed (to join to).

[0212] Example 3 - Specificity and relative affinity for MUC1-(S)Tn measured using FACS A panel of MUC1 IgG was screened on MCF7 cells expressing cancer-associated MUC1-Tn and MCF10A cells expressing wild-type MUC1. An analog of the reference antibody pancomab was included as a control for relative MUC1 expression in different cell lines. 293FF (MUC1-negative) cells were included as a negative assay control. Isotype control antibodies and analogs of the reference antibody 5E5 were included as negative and positive antibody controls, respectively. The MUC1 IgG panel and control antibodies were titrated using eight 4-fold serial dilutions starting at 10 ug / ml. Binding to 293FF cells was tested at 10 ug / ml.

[0213] MCF7 cells (DSMZ, ACC115), MCF10A cells (ATCC, catalog no. CRL-10317), or 293FF cells (Invitrogen, p / n51-0029) were incubated with MUC1 and FACS buffer (PBS + 0.5% BSA + 2 mM EDTA) with a control antibody diluted on ice for 30 minutes, followed by two washes with ice-cold FACS buffer and staining with anti-human IgG-PE (Invitrogen, #H10104) at 1:100. Binding was measured using iQue Screener PLUS. Analysis was performed using IntelliCyt ForeCyt® software. The mean fluorescence intensity (MFI) value of each MUC1 antibody was plotted against IgG concentration, and the area under the curve (AUC) was calculated based on normalization using isotype antibodies and analogues of reference antibody 5E5 as negative and positive antibody controls, respectively.

[0214] The results are shown in Figure 2. As expected, the positive and negative control antibodies bound to MCF7, MCF10A, and 293FF cells. The MUC1 core peptide conjugate (CB) bound to both MCF7 and MCF10A cells, with some showing low binding to 293FF cells. The MUC1-(S)Tn conjugate (SEQ ID NO) specifically bound to MCF7 but not to MCF10A cells. The MUC1-Tn conjugate (TnB) specifically bound to MCF7 but not to MCF10A cells, although it bound with lower affinity compared to the MUC1-(S)Tn conjugate. The affinity of the MUC1-(S)Tn conjugate to MCF7 cells varied, with many antibodies showing a higher binding signal than analogues of the reference antibody 5E5.

[0215] Example 4 - MUC1 measured using ELISA 40 Tn 10 and MUC1 40 STn 10 Affinity of the MUC1 Fab panel A panel of MUC1-(S)Tn IgG was subjected to GingisKHAN digestion to produce Fab (>99% monomer). Biotinylated MUC1 40 Tn 10 and MUC1 40 STn 10 The Fab fragments were captured on a 2 μg / ml nutravidine-coated plate. A Fab fragment of an analog of the reference antibody 5E5 was included as the positive control antibody, and a Fab fragment of the isotype control antibody was included as the negative control antibody.

[0216] The MUC1-(S)Tn Fab panel and control Fab were serially diluted using eight 10-fold serial dilutions starting from 10 ug / ml and incubated at room temperature for 1 hour. Subsequently, diluted HRP conjugate secondary antibody (Becton Dickinson, catalog number 555788) was incubated at room temperature for 1 hour. After each incubation step, three washing steps were performed using PBST. The reaction was stopped using 1 M H2SO4, and the signal was developed using TMB peroxidase substrate solution (BD Biosciences, catalog number 555214). The plates were then scanned using a microplate reader. 450 This was analyzed.

[0217] MUC1-(S)Tn Fab's MUC1 40 Tn 10 and MUC1 40 STn 10 The affinity for MUC1-Tn is similar to that for IgG (data not shown). A general correlation in affinity was found for MUC1-Tn peptide and MUC1-STn peptide (see Figure 3). Several clones show high affinity for MUC1-Tn and MUC1-STn. Within this group, many of the Fabs showing the best binding affinity for MUC1-Tn and MUC1-STn share the same HCDR1 sequence (circled in Figure 3).

[0218] Example 5 - Glycopeptide Array A panel of MUC1 IgG was screened using a glycopeptide array to determine their binding to the O-glycopeptides listed in Table 2. Print buffer was included as a negative assay control. Human IgG and mouse IgG were used as positive assay controls. Control antibodies included analogues of reference antibody 5E5, and analogues of reference antibodies VU4H5, 5F4, and 3F1. MUC1 and control antibodies were titrated using eight 4-fold serial dilutions starting at 10 ug / ml.

[0219] O-glycan array assays were performed according to manual instructions (Z Biotech, LLC). The array was blocked for 30 minutes using glycan array blocking buffer (GAAB, Z Biotech item number 10106). It was then washed three times with TBS-T based glycan array assay buffer (GAAB, Z Biotech item number 10107). MUC1 IgG and control antibody were diluted in GAAB in the range of 10 μg / ml to 0.6 ng / ml and then applied directly to the array. The array was covered with adhesive film and shaken at 80 rpm at room temperature for 1 hour. The array was then washed three times again with GAAB. Detection antibody, Cy3 conjugate anti-human IgG, and Cy3 conjugate anti-mouse IgG were diluted in GAAB and applied to the array. It was covered with light and shaken at 80 rpm at room temperature for 1 hour, then washed three times with GAAB and twice with MilliQ water. The array was read using an Innopsys InnoScan 710 microarray scanner with a 1PMT high-power laser. Software was used to detect each spot on the array and calculate the relative fluorescence unit (RFU) intensity for each spot. Background RFU was subtracted from the RFU value for each spot. The median RFU for each glycan spot was determined and graphed in the given report. [Table 2]

[0220] All control antibodies bound to the described epitopes. The results for MUC1 IgG are shown in Figure 4. MUC1 IgG, previously identified as a MUC1-(S)Tn binder, showed strong binding to peptides T4, T9, T11, S4, S9, and S11. All of these peptides contain either a modification of the threonine residue in the VTSA motif, a Tn modification (α-GalNAc glycosylation), or an STn modification (Neu5Ac-α(2-6)-GalNAc glycosylation). None of the other peptides contain a modification at this position, and the MUC1-(S)Tn binder does not bind to these peptides, except for peptide T8. The MUC1-(S)Tn binder also binds to peptides when, in addition to (S)Tn modification of threonine residues in the VTSA motif, the peptide contains Tn or STn modification of serine residues in the VTSA motif (T9, S9) or Tn or STn modification of threonine residues in the PDTR motif (T11, S11).

[0221] MUC1 IgG, previously identified as a MUC1-Tn binder, showed strong binding to peptides T2, T7, and T10. All of these peptides contain Tn modification (α-GalNAc glycosylation) of a serine residue in the GSTA motif. None of the other peptides contain modification at this position, and the MUC1-(S)Tn binder does not bind to those peptides. If, in addition to Tn modification of a serine residue in the GSTA motif, the peptide also contains Tn modification of a threonine residue in the GSTA motif (T7), or Tn modification of a threonine residue in the GSTA motif and Tn modification of a threonine residue in the PDTR motif (T10), the MUC1-Tn binder will also bind to the peptide.

[0222] Example 6 - Affinity of MUC1 IgG for T47D huGalNAc cells MUC1 IgG was screened for binding to T47D huGalNAc cells stably expressing human MUC1-STn and T47D WT cells expressing moderate levels of MUC-TN. Isotype control antibodies were included as negative controls, and an analog of reference antibody 5E5 was included as a positive control. An analog of the reference antibody pancomab was included as a control for the relative expression of MUC1 in different cell lines. Further control antibodies included 3F1 and 5F4. MUC1, isotype controls, the 5E5 analog, and the pancomab analog were titrated using eight sequential 4-fold dilutions starting at 10 ug / ml, with 3F1 and 5F4 used at 10 ug / ml.

[0223] T47D cells (Sigma-Aldrich, catalog number 85102201-1VL) and T47D huGalNAc cells (generated by stably transfecting T47D cells with a plasmid encoding ST6GALNAC1 (acceptance number Q9NSC7; Uniprot)) were incubated with MUC1 IgG and control antibodies diluted on ice for 30 minutes in FACS buffer (PBS + 0.5% BSA + 2 mM EDTA), followed by two washes with ice-cold FACS buffer. The cells were then stained with anti-human IgG-PE (Invitrogen, catalog number H10104) at a ratio of 1:100 (for MUC1, isotype control, 5E5 analogue, and pancomab antibody sample analogue) or anti-mouse IgG-PE (Invitrogen, catalog number M30004-4) at a ratio of 1:400 (for 3F1 and 5F4 antibody samples). Binding was measured using iQue Screener PLUS. Analysis was performed using IntelliCyt ForeCyt® software. The MFI values ​​of each MUC1 antibody were plotted against IgG concentration, and the area under the curve (AUC) was calculated based on normalization using isotype control antibodies and analogues of the reference antibody 5E5 as negative and positive antibody controls, respectively.

[0224] The results are shown in Figure 5. The panel of MUC1(S)Tn conjugates exhibits diverse binding affinities to T47D huGalNAc cells. Several antibodies show higher maximum binding than analogues of the reference antibody 5E5. Affinity for T47D huGalNAc cells correlates well with affinity for the MUC1-STn peptide.

[0225] A panel of MUC1-(S)Tn conjugates exhibits diverse binding affinities to T47D WT cells. Most antibodies show higher maximum binding than analogues of reference antibody 5E5, corresponding to data obtained in MCF7 cells. Affinity for T47D WT cells is MUC1 40 Tn 10 It correlates even better with affinity for peptides.

[0226] Example 7 - Affinity of MUC1 IgG for MC38 cells MUC1 IgG was screened for binding to MC38 huMUC1 COSMC KO cells expressing human MUC1-Tn, MC38 huMUC1 cells expressing normal human MUC1 glycosphatids, and MC38 WT cells that do not express human MUC1. Isotype control antibodies were included as negative controls, and an analog of reference antibody 5E5 was included as a positive control. An analog of the reference antibody pancomab was included as a control for the relative expression of MUC1 in different cell lines. Further control antibodies included 3F1 and 5F4. MUC1 IgG, isotype controls, 5E5 analogs, and pancomab analogs were titrated using eight sequential 4-fold dilutions starting at 10 ug / ml, with 3F1 and 5F4 used at 10 ug / ml.

[0227] MC38 huMUC1 COSMC KO cells were generated by stably transfecting MC38 huMUC1 cells (acceptance number: CVCL_5l38; Uniprot) with human MUC1 and knocking out the mouse COSMC (C1galt1c1) gene (acceptance number: ENSMUST00000058265.7; Ensemble). Cells were incubated with MUC1 IgG or control antibody diluted in FACS buffer (PBS + 0.5% BSA + 2 mM EDTA) on ice for 30 minutes, followed by two washes with ice-cold FACS buffer. Secondary antibodies were stained at 1:100 with anti-human IgG-PE (Invitrogen, catalog no. H10104) (for MUC1, isotype control, 5E5 analogue, and pancomab antibody sample analogue) or 1:400 with anti-mouse IgG-PE (Invitrogen, catalog no. M30004-4) (for 3F1 and 5F4 antibody samples). Binding was measured using iQue Screener PLUS. Analysis was performed using IntelliCyt ForeCyt® software. The MFI values ​​of each huMUC1-(S)Tn clone were plotted against IgG concentration, and the area under the curve (AUC) was calculated based on normalization using isotype control antibodies and analogues of the reference antibody 5E5 as negative and positive antibody controls, respectively.

[0228] The results are shown in Figure 6. Positive and negative control antibodies function as expected in MC38 huMUC1 COSMC KO cells, MC38 huMUC1 cells, and MC38 WT cells. Except for the antibody containing the heavy chain variable region with SEQ ID NO: 178, the MUC1 antibodies did not bind to MC38 WT cells. However, this antibody did not bind to the CALR WT negative control peptide or 293FF WT cells. The panel of MUC1-(S)Tn conjugates showed a high binding signal to MC38 huMUC1 COSMC KO cells. The panel of MUC1-(S)Tn conjugates showed a very low binding signal to MC38 huMUC1 cells, indicating specificity for MUC1-(S)Tn. array Sequence ID 1 - Heavy Chain Variable Region QVQLVESGAEVKKPGASVRLSCKASGYSFIHYYLHYVRQAPGQGLEWMGVINPFDGSTDYAQKFHGRVTLTRDTSASTVFMELSSLGSEDTAVYFCTRDNMIGYVLFDYWGLGTLVTVSS Sequence ID 2 - Heavy Chain CDR1 HYYLH Sequence ID 3 - Heavy Chain CDR2 VINPFDGSTDYAQKFHG Sequence ID 4 - Heavy Chain CDR3 DNMIGYVLFDY Sequence ID 5 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKTSGYTFTRYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQKFQGRVTVTRDTSTFTVYMELSSLRSEDTAVYYCARDYMRGYVIFDYWGQGTLVTVSS Sequence ID 6 - Heavy Chain CDR1 RYYIQ Sequence ID 7 - Heavy Chain CDR2 VINPYDGSTNYAQKFQG Sequence ID 8 - Heavy Chain CDR3 DYMRGYVIFDY Sequence ID 9 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFNRHYIQWVRQAPGQGLEWMGTINPYDGSTHYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFDYWGQGTLVTVSS Sequence ID 10 - Heavy Chain CDR1 RHYIQ Sequence ID 11 - Heavy Chain CDR2 TINPYDGSTHYAQKFQG Sequence ID 12 - Heavy Chain CDR3 DYMIGYVVFDY Sequence ID 13 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASLKVSCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQKFQGRVTLTRDTSTTTVYMELSSLRSDDTAVYYCARDYMIGYVVFDYWGQGTLVTVSS Sequence ID 14 - Heavy Chain CDR1 NYYIQ Sequence ID 15 - Heavy Chain CDR2 VINPYDGSTNYAQKFQG Sequence ID 16 - Heavy Chain CDR3 DYMIGYVVFDY Sequence ID 17 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVRVSCKASGYPFINYYIQWVRQAPGQGLEWMGVVNPYDGSTNYAQKFQGRVSITRDASTSTVFMELNSLRSEDTAVYYCARDSMLGHVIFDFWGQGTLVTVSS Sequence ID 18 - Heavy Chain CDR1 NYYIQ Sequence ID 19 - Heavy Chain CDR2 VVNPYDGSTNYAQKFQG Sequence ID 20 - Heavy Chain CDR3 DSMLGHVIFDF Sequence ID 21 - Heavy Chain Variable Region QVQLVQSGAEVRKPGASVKFSCKASGYSFTHYYVHWVRQAPGQGLEWMGVINPFDSSTNYARKFQGRVTMTRDTSTTTVYMDLSSLRSEDTAVYFCARDAMEGDSYFDYWGQGTLVTVSS Sequence ID 22 - Heavy Chain CDR1 HYYVH Sequence ID 23 - Heavy Chain CDR2 VINPFDSSTNYARKFQG Sequence ID 24 - Heavy Chain CDR3 DAMEGDSYFDY Sequence ID 25 - Heavy Chain Variable Region QVQLVESGAEVKKPGASVKVSCKASGYTFARYYVHWVRQAPGQGLEWMGLINPYGGATNYAQNFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDAMGGDSYFDYWGQGTLVTVSS Sequence ID 26 - Heavy Chain CDR1 RYYVH Sequence ID 27 - Heavy Chain CDR2 LINPYGGATNYAQNFQG Sequence ID 28 - Heavy Chain CDR3 DAMGGDSYFDY Sequence ID 29 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYSFIHYYIHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQDRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDDWGQGTLVTVSS Sequence ID 30 - Heavy Chain CDR1 HYYIH Sequence ID 31 - Heavy Chain CDR2 VINPFDGSTNYAQKFQD Sequence ID 32 - Heavy Chain CDR3 DNMIGYVLFDD Sequence ID 33 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVRVSCKASGYPFINYYIQWVRQAPGQGLEWMGVINPFDGSTNYAQKFQDRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLLDYWGQGTLVTVSS Sequence ID 34 - Heavy Chain CDR1 NYYIQ Sequence ID 35 - Heavy Chain CDR2 VINPFDGSTNYAQKFQD Sequence ID 36 - Heavy Chain CDR3 DNMIGYVLLDY Sequence ID 37 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVTVSCKTSGYTFTSYYIHWLRQAPGQGLEWMGVINPFGGATNYAQKFQGRVTMTRDTSTITVYMELSSLRSEDTAVYYCARDAMGGDSHFDHWGRGTLVTVSS Sequence ID 38 - Heavy Chain CDR1 SYYIH Sequence ID 39 - Heavy Chain CDR2 VINPFGGATNYAQKFQG Sequence ID 40 - Heavy Chain CDR3 DAMGGDSHFDH Sequence ID 41 - Heavy Chain Variable Region QVQLVQSGAEVKEPGASMTLSCKTSGYTFISNYIHWLRQAPGQGLEWLGVINPFGGATNYAQKFQGRVTMTRDTSTITVYMELSSLRSEDTAVYYCARDAMGGDSHFDYWGQGTLVTVSS Sequence ID 42 - Heavy Chain CDR1 SNYIH Sequence ID 43 - Heavy Chain CDR2 VINPFGGATNYAQKFQG Sequence ID 44 - Heavy Chain CDR3 DAMGGDSHFDY Sequence ID 45 - Heavy Chain Variable Region QVQLVESGTEVKKPGASVKVSCKASGYSFIHYYIHWVRQAPGRGLEWMGVINPFDGSTNSAQNFQGRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMLGYVLFDHWGQGTLVTVSS Sequence ID 46 - Heavy Chain CDR1 HYYIH Sequence ID 47 - Heavy Chain CDR2 VINPFDGSTNSAQNFQG Sequence ID 48 - Heavy Chain CDR3 DNMLGYVLFDH Sequence ID 49 - Heavy Chain Variable Region QVQLVESGAEVKKPGASVKVSCKASGYTFTRYYLHWVRQAPGQGLEWMGVINPYGGATNYAQKFQGRVTLTRDTSTSTVYMELNSLRSEDTAVYFCARDAMEGNSYFDHWGPGTLVTVSS Sequence ID 50 - Heavy Chain CDR1 RYYLH Sequence ID 51 - Heavy Chain CDR2 VINPYGGATNYAQKFQG Sequence ID 52 - Heavy Chain CDR3 DAMEGNSYFDH Sequence ID 53 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASMTVSCKTSGYTFISYYIHWLRQAPGQGLEWLGVINPFGGATNYAQKFQGRVTMTRDTSTITVYMELSSLRSEDTAVYYCARDAMGGDSHFDFWGQGTLVTVSS Sequence ID 54 - Heavy Chain CDR1 SYYIH Sequence ID 55 - Heavy Chain CDR2 VINPFGGATNYAQKFQG Sequence ID 56 - Heavy Chain CDR3 DAMGGDSHFDF Sequence ID 57 - Heavy Chain Variable Region QVQLVESGTEVKKPGASVKVSCKASGYSFIHYYIHWVRQAPGRGLEWMGVINPFDGSTNYAQNFQGRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDHWGQGTLVTVSS Sequence ID 58 - Heavy Chain CDR1 HYYIH Sequence ID 60 - Heavy Chain CDR2 VINPFDGSTNYAQNFQG Sequence ID 61 - Heavy Chain CDR3 DNMIGYVLFDH Sequence ID 62 - Heavy Chain Variable Region QVQLVESGAEVKKPGASVKVSCKASGYSFTSYYIQWVRQAPGQGLEWLAVINPIDGSTNYAQKFQGRVTVTRDTSTSTVYMELSSLRSEDTAVYYCARDYMRGFVVFDYWGQGTLVTVSS Sequence ID 63 - Heavy Chain CDR1 SYYIQ Sequence ID 64 - Heavy Chain CDR2 VINPIDGSTNYAQKFQG Sequence ID 65 - Heavy Chain CDR3 DYMRGFVVFDY Sequence ID 66 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVRVSCKASGYTFTNYYIHWVRQAPGHGLTWMGVINPFDSSTDYAQKFQGRVTMTRDTSTSTVYMDLSSLRSEDTAVYYCARDYMRGHVVFDYWGQGTLVTVSS Sequence ID 67 - Heavy Chain CDR1 NYYIH Sequence ID 68 - Heavy Chain CDR2 VINPFDSSTDYAQKFQG Sequence ID 69 - Heavy Chain CDR3 DYMRGHVVFDY Sequence ID 70 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYAFPNYYIHWVRQAPGQGLEWMGVINPYGGSTNYAQKFQGRVTMTRGTSTSTVYLELSSLRSEDTAVYYCARDAMKGDSFFDYWGQGTLVTVSS Sequence ID 71 - Heavy Chain CDR1 NYYIH Sequence ID 72 - Heavy Chain CDR2 VINPYGGSTNYAQKFQG Sequence ID 73 - Heavy Chain CDR3 DAMKGDSFFDY Sequence ID 74 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVQVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYEQNFQGRVTMTRDTSTNTVYMELNGLRSEDTAVYYCARDHMIGYILFDYWGQGTTVTVSS Sequence ID 75 - Heavy Chain CDR1 NYYLQ Sequence ID 76 - Heavy Chain CDR2 TINPFDGSTHYEQNFQG Sequence ID 77 - Heavy Chain CDR3 DHMIGYILFDY Sequence ID 78 - Heavy Chain Variable Region QVQLVQSGAEVKKPGTSVKVSCKASGYSFIHYYIHWVRQAPGQGLEWVGVINPFDGSTNYAQKFQGRVTVTRDTSTSAVFMDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS Sequence ID 79 - Heavy Chain CDR1 HYYIH Sequence ID 80 - Heavy Chain CDR2 VINPFDGSTNYAQKFQG Sequence ID 81 - Heavy Chain CDR3 DNMIGYVLFDY Sequence ID 82 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIHWVRQAPGQGLEWMGVINPYDSSTNYAQKFQGRVTMTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS Sequence ID 83 - Heavy Chain CDR1 SYYIH Sequence ID 84 - Heavy Chain CDR2 VINPYDSSTNYAQKFQG Sequence ID 85 - Heavy Chain CDR3 DNMIGYVLFDY Sequence ID 86 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYPFINYYIQWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFMDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS Sequence ID 87 - Heavy Chain CDR1 NYYIQ Sequence ID 88 - Heavy Chain CDR2 VINPFDGSTNYAQKFQG Sequence ID 89 - Heavy Chain CDR3 DNMIGYVLFDY Sequence ID 90 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFINHYIHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFLDLSSLRSEDTAVYYCARDNMIGYVLFDYWGQGTLVTVSS Sequence ID 91 - Heavy Chain CDR1 NHYIH Sequence ID 92 - Heavy Chain CDR2 VINPFDGSTNYAQKFQG Sequence ID 93 - Heavy Chain CDR3 DNMIGYVLFDY Sequence ID 94 - Heavy Chain Variable Region QVQLVQSGAEVKKPGSSVKVSCKASGYTFNSYYIHWVRQAPGQGLTWMGVINPFDGSTDYAQKFQGRVTMTRDTSTSTVYMDLSSLRSEDTAVYYCARDYMRGYVVFDYWGRGTLVTVSS Sequence ID 95 - Heavy Chain CDR1 SYYIH Sequence ID 96 - Heavy Chain CDR2 VINPFDGSTDYAQKFQG Sequence ID 97 - Heavy Chain CDR3 DYMRGYVVFDY Sequence ID 98 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYIQWLRQAPGQGLEWMGVINPTGGATNYAQKFQGRVTMTRETSTSMVHMELSSLRSEDTAVYYCARDYMRGYVVFDYWGQGTLVTVSS Sequence ID 99 - Heavy Chain CDR1 SYYIQ Sequence ID 100 - Heavy Chain CDR2 VINPTGGATNYAQKFQG Sequence ID 101 - Heavy Chain CDR3 DYMRGYVVFDY Sequence ID 102 - Heavy Chain Variable Region QVQLVQSGAEVKKSGASVKVSCKASGYPFISYYIHWLRQAPGQGLTWMGVINPFDGSTDYAQKFQGRVTVTRDTSTSTVYMDLSSLRADDTAIYYCARDYMRGYVVFDYWGQGTLVTVSS Sequence ID 103 - Heavy Chain CDR1 SYYIH Sequence ID 104 - Heavy Chain CDR2 VINPFDGSTDYAQKFQG Sequence ID 105 - Heavy Chain CDR3 DYMRGYVVFDY Sequence ID 106 - Heavy Chain Variable Region QVQLVQSGAEMKKPGASVKISCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQRFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFDFWGQGTLVTVSS Sequence ID 107 - Heavy Chain CDR1 NYYIQ Sequence ID 108 - Heavy Chain CDR2 VINPYDGSTNYAQRFQG Sequence ID 109 - Heavy Chain CDR3 DYMIGYVVFDF Sequence ID 110 - Heavy Chain Variable Region QVQLVQSGAEMKKPGASVKVSCKASGYTFTNYYIHWVRQAPGQGLEWMGVINPYDGSTNYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFDFWGQGTLVTVSS Sequence ID 111 - Heavy Chain CDR1 NYYIH Sequence ID 112 - Heavy Chain CDR2 VINPYDGSTNYAQKFQG Sequence ID 113 - Heavy Chain CDR3 DYMIGYVVFDF Sequence ID 114 - Heavy Chain Variable Region QVQLVQSGAEMKKPGASVTVSCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQRFQGRVTLTRDTTTTTVNMELSGLRSEDTAVYYCARDYMIGYVVFDFWGQGTLVTVSS Sequence ID 115 - Heavy Chain CDR1 NYYIQ Sequence ID 116 - Heavy Chain CDR2 VINPYDGSTNYAQRFQG Sequence ID 117 - Heavy Chain CDR3 DYMIGYVVFDF Sequence ID 118 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYYIQWVRQAPGQGLEWMGVINPYDGSTNYAQRFQGKITLTRDTSTTTVYMELSSLGSEDTAVYYCARDYMIGYVVFDYWGQGTMVTVSS Sequence ID 119 - Heavy Chain CDR1 NYYIQ Sequence ID 120 - Heavy Chain CDR2 VINPYDGSTNYAQRFQG Sequence ID 121 - Heavy Chain CDR3 DYMIGYVVFDY Sequence ID 122 - Heavy Chain Variable Region EVQLLESGAEVKKPGASVKVSCKASGYTFNRYYIHWVRQAPGQGLEWMGVINPYDGSTDYAQKFRDRITMTRDTSTNTVYMDLSSLRSEDTAVYFCARDAMEGDSYFDNWGRGTLVTVSS Sequence ID 123 - Heavy Chain CDR1 RYYIH Sequence ID 124 - Heavy Chain CDR2 VINPYDGSTDYAQKFRD Sequence ID 125 - Heavy Chain CDR3 DAMEGDSYFDN Sequence ID 126 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFNRYYLHWVRQAPGQGLEWMGVINPYDGSTNYAQKFRGRVTMTRDTSTNTVYMELSSLRSEDTAVYFCARDAMEGDSYFDNWGQGTTVTVSS Sequence ID 127 - Heavy Chain CDR1 RYYLH Sequence ID 128 - Heavy Chain CDR2 VINPYDGSTNYAQKFRG Sequence ID 129 - Heavy Chain CDR3 DAMEGDSYFDN Sequence ID 130 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVRVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYAQKFQGRVTLTRDTSTSTVYMELNSLRSDDTAVYYCARDYMIGYVLFDYWGQGTLVTVSS Sequence ID 131 - Heavy Chain CDR1 NYYLQ Sequence ID 132 - Heavy Chain CDR2 TINPFDGSTHYAQKFQG Sequence ID 133 - Heavy Chain CDR3 DYMIGYVLFDY Sequence ID 134 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVEVSCKASGYTFINYYIHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFLDLSSLRPEDTAVYYCARDSMIGYVLFDYWGQGTLVTVSS Sequence ID 135 - Heavy Chain CDR1 NYYIH Sequence ID 136 - Heavy Chain CDR2 VINPFDGSTNYAQKFQG Sequence ID 137 - Heavy Chain CDR3 DSMIGYVLFDY Sequence ID 138 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVRVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARDSMIGYVLFDYWGQGTLVTVSS Sequence ID 139 - Heavy Chain CDR1 NYYLQ Sequence ID 140 - Heavy Chain CDR2 TINPFDGSTHYAQKFQG Sequence ID 141 - Heavy Chain CDR3 DSMIGYVLFDY Sequence ID 142 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVMLSCKASGYIFTSYYIQWVRQAPGQGLEWMGVVNPYDGSTNYAQKFQGRVSITRDTSTSTVFMELNSLRSEDTAVYYCARDSMLGHVIFDFWGQGTLVTVSS Sequence ID 143 - Heavy Chain CDR1 SYYIQ Sequence ID 144 - Heavy Chain CDR2 VVNPYDGSTNYAQKFQG Sequence ID 145 - Heavy Chain CDR3 DSMLGHVIFDF Sequence ID 146 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYAFTSYYIHWVRQAPGQGLEWMGVINPYGGSTNYAQKFQGRVTMTRDTSTRIVYMDLSSLRSEDTAVYYCARDAMEGDSYFDYWGQGTLVTVSS Sequence ID 147 - Heavy Chain CDR1 SYYIH Sequence ID 148 - Heavy Chain CDR2 VINPYGGSTNYAQKFQG Sequence ID 149 - Heavy Chain CDR3 DAMEGDSYFDY Sequence ID 150 - Heavy Chain Variable Region QVQLVQSGAEVKTPGASVKVSCKASGYTFTNYYIQWVRQAPGQALEWMGVINPYDGSTHYAQKFQGRVTLTRDTSTTTVYMDLSSLRSEDAAVYYCARDYMLGYVVFDYWGQGTLVTVSS Sequence ID 151 - Heavy Chain CDR1 NYYIQ Sequence ID 152 - Heavy Chain CDR2 VINPYDGSTHYAQKFQG Sequence ID 153 - Heavy Chain CDR3 DYMLGYVVFDY Sequence ID 154 - Heavy Chain Variable Region QVQLVQSGAEVKKSGASVKVSCKASGYIFTSYYIQWVRQAPGQGLEWMGVINPFDGSTDYAQKFQGRVTVTRDTSTTTVFMELSSLRSEDTAVYYCARDNMLGYVLFDYWGQGTLVTVSS Sequence ID 155 - Heavy Chain CDR1 SYYIQ Sequence ID 156 - Heavy Chain CDR2 VINPFDGSTDYAQKFQG Sequence ID 157 - Heavy Chain CDR3 DNMLGYVLFDY Sequence ID 158 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTATTTVFMDLSSLRSEDTAVYYCARDNMLGYVLFDYWGQGTLVTVSS Sequence ID 159 - Heavy Chain CDR1 SYYMH Sequence ID 160 - Heavy Chain CDR2 VINPFDGSTNYAQKFQG Sequence ID 161 - Heavy Chain CDR3 DNMLGYVLFDY Sequence ID 162 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVMLSCKASGYIFTSYYIQWVRQAPGQGLEWMGVVNPYDGSTNYAQKFQGRVTITRDTSTSTVFMELSSLRSEDTAVYYCARDNMLGHVIFDFWGQGTLVTVSS Sequence ID 163 - Heavy Chain CDR1 SYYIQ Sequence ID 164 - Heavy Chain CDR2 VVNPYDGSTNYAQKFQG Sequence ID 165 - Heavy Chain CDR3 DNMLGHVIFDF Sequence ID 166 - Heavy Chain Variable Region QVQLVESGAEVKKPGASVKVSCKASGYTFNRYYIHWVRQAPGQGLEWMGVINPSDGSTNYAQKFRGRITMTRDTSTNIVYMELSSLRSEDTAVYFCARDAMGGDSYFDNWGQGTLVTVSS Sequence ID 167 - Heavy Chain CDR1 RYYIH Sequence ID 168 - Heavy Chain CDR2 VINPSDGSTNYAQKFRG Sequence ID 169 - Heavy Chain CDR3 DAMGGDSYFDN Sequence ID 170 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVQVSCKASGYTFINYYLQWVRQAPGQGLEWMGTINPFDGSTHYAQKFQGRVTMTRDTSTNTVYMELNSLRSEDTAVYYCARDRMIGYVLFDYWGQGTLVTVSS Sequence ID 171 - Heavy Chain CDR1 NYYLQ Sequence ID 172 - Heavy Chain CDR2 TINPFDGSTHYAQKFQG Sequence ID 173 - Heavy Chain CDR3 DRMIGYVLFDY Sequence ID 174 - Heavy Chain Variable Region EVQLVESGAEVKKPGASVKVSCKASGYTFNRYYLHWVRQAPGQGLEWMGVINPYDGSTNYAQKFRGRITMTRDTSTNTVYMELSSLRSEDTAVYFCARDAMEGDSYFDKWGQGTLVTVSS Sequence ID 175 - Heavy Chain CDR1 RYYLH Sequence ID 176 - Heavy Chain CDR2 VINPYDGSTNYAQKFRG Sequence ID 177 - Heavy Chain CDR3 DAMEGDSYFDK Sequence ID 178 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFNRHYIQWVRQAPGQGLEWMGTINPYDGSTFYAQKFQDRVTMTRDTSTSTVYMDLNSLKSEDTAVYYCARDYMRGHVLFDYWGQGTLVTVSS Sequence ID 179 - Heavy Chain CDR1 RHYIQ Sequence ID 180 - Heavy Chain CDR2 TINPYDGSTFYAQKFQD Sequence ID 181 - Heavy Chain CDR3 DYMRGHVLFDY Sequence ID 182 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYSFTYSYIQWVRLAPGRGLEWMGVINPYDGSTDYAQKFQGRVTLTRDTSTTTVYMELSSLTSEDTAVYYCARDFMIGYVVFDYWGQGTLVTVSS Sequence ID 183 - Heavy Chain CDR1 YSYIQ Sequence ID 184 - Heavy Chain CDR2 VINPYDGSTDYAQKFQG Sequence ID 185 - Heavy Chain CDR3 DFMIGYVVFDY Sequence ID 186 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFNRYYIHWVRQAPGQGLEWMGVINPYDGSTNYAQKFRGRITMTRDTSTNTVYMDLSSLRSEDTAVYFCARDAMEGDSYLDNWGRGTLVTVSS Sequence ID 187 - Heavy Chain CDR1 RYYIH Sequence ID 188 - Heavy Chain CDR2 VINPYDGSTNYAQKFRG Sequence ID 189 - Heavy Chain CDR3 DAMEGDSYLDN Sequence ID 190 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYAFTNYYIHWVRQAPGHGLTWMGVINPFDSSTDFAQKFQGRLTMTRDTSTSTVYMDLSSLRSEDTAVYYCARDYMRGHVVFDNWGQGTLVTVSS Sequence ID 191 - Heavy Chain CDR1 NYYIH Sequence ID 192 - Heavy Chain CDR2 VINPFDSSTDFAQKFQG Sequence ID 193 - Heavy Chain CDR3 DYMRGHVVFDN Sequence ID 194 - Heavy Chain Variable Region QVQLVQSGAEVKKPGTSVKVSCTASGYTFTNYYIQWVRQAPGQGLEWLGVINPYDGSTDYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMLGYVVFDFWGQGTLVTVSS Sequence ID 195 - Heavy Chain CDR1 NYYIQ Sequence ID 196 - Heavy Chain CDR2 VINPYDGSTDYAQKFQG Sequence ID 197 - Heavy Chain CDR3 DYMLGYVVFDF Sequence ID 198 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVKVSCKASGYTFINNYIQWVRQAPGQGLEWMGVINPYDGSIHYAQKFQGRVTLTRDTSTTTVYMELSSLRSEDTAVYYCARDYMIGYVVFEYWGQGTLVTVSS Sequence ID 199 - Heavy Chain CDR1 NNYIQ Sequence ID 200 - Heavy Chain CDR2 VINPYDGSIHYAQKFQG Sequence ID 201 - Heavy Chain CDR3 DYMIGYVVFEY Sequence ID 202 - Heavy Chain Variable Region QVQLVQSGAEVKNPGASVKFSCKASGYTFINYYLQWLRQAPGQGLEWMGTINPFDGSTSYAQNFQGRVTMTRDTSSTTVYMELNSLKSEDTAVYYCARDSMIGYVLFDSWGQGTLVTVSS Sequence ID 203 - Heavy Chain CDR1 NYYLQ Sequence ID 204 - Heavy Chain CDR2 TINPFDGSTSYAQNFQG Sequence ID 205 - Heavy Chain CDR3 DSMIGYVLFDS Sequence ID 206 - Heavy Chain Variable Region QVQLVQSGAEVKKPGASVRVSCKASGYPFINYYIQWVRQAPGQGLEWMGVINPFDGSTNYAQKFQGRVTVTRDTSTSTVFMDLNSLRSEDTAVYYCARDSMIGYVLFDFWGQGTLVTVSS Sequence ID 207 - Heavy Chain CDR1 NYYIQ Sequence ID 208 - Heavy Chain CDR2 VINPFDGSTNYAQKFQG Sequence ID 209 - Heavy Chain CDR3 DSMIGYVLFDF Sequence ID 210 - Heavy Chain 5E5 Analog QVQLVQSGAEVKKTGSSVKVSCKASGYTFTDHAIHWVRQAPGQALEWMGHFSPGNTDIKYNDKFKGRVTLTVDRSMSTAYMELSSLRSEDTAMYYCKTSTFFFDYWGQGTM VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTH TCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence ID 211 - Light chain 5E5 analog DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGDQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYSYPLTFGQGTKV EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 212 - Heavy chain pancomab analog EVKLVESGGGLVQPGGSMKLSCVASGFTFSNYWMNWVRQSPEKGLEWVAEIRLKSNNYTTHYAESVKGRFTISRDDSKSSVSLQMNNLRVEDTGIYYCTRHYYFDYWGQGT TVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKT HTCPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence ID 213 - Light chain pancomab analog DIVMTQAAFSNPVTLGTSASISCRSSKSLLHSNGITYFFWYLQKPGLSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLELPPTFGGGTKL EIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 214 - Heavy chain HT186-D11 analog QMQLVQSEAELKKPGASVKVSCKASGYSFTGHYMHWVRQAPGQGLEWMGWIDPVTGGTKYAQNFQGWVTMTRDTSIRTAYLELSRLRSDDTAMYYCAREVTGDRGQFDKWGQ GTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence ID 215 - Light chain HT186-D11 analog QSVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPALVIYYGSNRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDWVFGGGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 216 - Heavy Chain Isotype Control IgG EVQLVETGAEVKKPGASVKVSCKASDYIFTKYDINWVRQAPGQGLEWMGWMSANTGNTGYAQKFQGRVTMTRDTSINTAYMELSSLTSGDTAVYFCARSSLFKTETAPYYHFAL DVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPK SCDKTHTCPPCPAPELGRGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI EKTISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Sequence ID 217 - Light chain isotype control IgG DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 218 - Light chain variable region DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIK Light chain CDR1 by sequence number 219-IMGT QSISSY Light chain CDR2 by Sequence ID No. 220-IMGT AAS Light chain CDR3 by Sequence ID No. 221-IMGT QQSYSTPPT Sequence ID 222 - Hinge Region EPKSCDKTHTCPPCP Sequence ID 223-CH1 region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRV Sequence ID 224-CH2 region APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK Sequence ID 225-CH3 region GQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG Sequence ID 226 - CL region RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Sequence ID 227-V region VK1-39 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTP Sequence ID 228-VK1-39 / JK1 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPTFGQGTKVEIK Sequence ID 229-VK1-39 / JK5 DIQMTQSPSSLSASVGDRVTITCRASQSISSYLNWYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQSYSTPPITFGQGTRLEIK Light chain CDR1 by sequence number 230-IMGT QSISSY Light chain CDR2 by Sequence ID No. 231-IMGT AAS Light chain CDR3 by Sequence ID No. 232-IMGT QQSYSTPPIT Sequence ID 233-V region VK3-15 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWP Sequence ID 234-VK3-15 / JK1 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYNNWPWTFGQGTKVEIK Light chain CDR1 by sequence number 235-IMGT QSVSSN Light chain CDR2 by Sequence ID No. 236-IMGT GAS Light chain CDR3 by Sequence ID No. 237-IMGT QQYNNWPWT Sequence ID 238-V region VK3-20 EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSP Sequence ID 239-VK3-20 / JK1 EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGTKVEIK Light chain CDR1 by sequence number 240-IMGT QSVSSSY Light chain CDR2 by Sequence ID No. 241-IMGT GAS Light chain CDR3 by Sequence ID No. 242-IMGT QQYGSSPWT Sequence ID 243 - V region VL3~21 SYVLTQPPSVSVAPGETARITCGGDNIGRKSVYWYQQKSGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDGSSDH Sequence ID 244-VL3-21 / JL3 SYVLTQPPSVSVAPGETARITCGGDNIGRKSVYWYQQKSGQAPVLVIYYDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDGSSDHWVFGGGTKLTVL Light chain CDR1 by sequence number 245-IMGT NIGRKS Light chain CDR2 by Sequence ID No. 246-IMGT YDS Light chain CDR3 by Sequence ID No. 247-IMGT QVWDGSSDHWV Light chain CDR3 by Sequence ID No. 248-IMGT QQSYSTPPT Sequence ID 249 PAPGSTAPPAHGVTSAPDTRPAPG

Claims

1. A binding domain having binding specificity to cancer-related MUC1, wherein the binding domain is a) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), each having the amino acid sequence shown in SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, b) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), each having the amino acid sequence 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), each having the amino acid sequence 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), each having the amino acid sequence 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), each having the amino acid sequence shown in SEQ ID NO: 18, SEQ ID NO: 19, and SEQ ID NO: 20, f) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 22, SEQ ID NO: 23, and SEQ ID NO:

24. g) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), each having the amino acid sequence shown in SEQ ID NO: 26, SEQ ID NO: 27, and SEQ ID NO: 28, h) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 30, SEQ ID NO: 31, and SEQ ID NO:

32. i) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), each having the amino acid sequence shown in SEQ ID NO: 34, SEQ ID NO: 35, and SEQ ID NO: 36, j) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 38, SEQ ID NO: 39, and SEQ ID NO:

40. k) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 42, SEQ ID NO: 43, and SEQ ID NO:

44. l) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 46, SEQ ID NO: 47, and SEQ ID NO:

48. m) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 50, SEQ ID NO: 51, and SEQ ID NO:

52. n) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 54, SEQ ID NO: 55, and SEQ ID NO:

56. o) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 58, SEQ ID NO: 60, and SEQ ID NO:

61. p) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 63, SEQ ID NO: 64, and SEQ ID NO:

65. q) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 67, SEQ ID NO: 68, and SEQ ID NO:

69. r) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 71, SEQ ID NO: 72, and SEQ ID NO:

73. s) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 75, SEQ ID NO: 76, and SEQ ID NO:

77. t) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 79, SEQ ID NO: 80, and SEQ ID NO:

81. u) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 83, SEQ ID NO: 84, and SEQ ID NO:

85. v) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), each having the amino acid sequence 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), respectively, having the amino acid sequences shown in SEQ ID NO: 91, SEQ ID NO: 92, and SEQ ID NO:

93. x) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 95, SEQ ID NO: 96, and SEQ ID NO:

97. y) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 99, SEQ ID NO: 100, and SEQ ID NO:

101. z) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 103, SEQ ID NO: 104, and SEQ ID NO:

105. aa) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 107, SEQ ID NO: 108, and SEQ ID NO:

109. bb) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 111, SEQ ID NO: 112, and SEQ ID NO:

113. cc) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 115, SEQ ID NO: 116, and SEQ ID NO:

117. dd) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 119, SEQ ID NO: 120, and SEQ ID NO:

121. ee) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 123, SEQ ID NO: 124, and SEQ ID NO:

125. ff) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 127, SEQ ID NO: 128, and SEQ ID NO:

129. gg) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 131, SEQ ID NO: 132, and SEQ ID NO:

133. hh) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 135, SEQ ID NO: 136, and SEQ ID NO:

137. ii) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 139, SEQ ID NO: 140, and SEQ ID NO:

141. jj) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 143, SEQ ID NO: 144, and SEQ ID NO:

145. kk) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 147, SEQ ID NO: 148, and SEQ ID NO:

149. ll) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 151, SEQ ID NO: 152, and SEQ ID NO:

153. mm) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), each having the amino acid sequence 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), respectively, having the amino acid sequences shown in SEQ ID NO: 159, SEQ ID NO: 160, and SEQ ID NO:

161. oo) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 163, SEQ ID NO: 164, and SEQ ID NO:

165. pp) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 167, SEQ ID NO: 168, and SEQ ID NO:

169. qq) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 171, SEQ ID NO: 172, and SEQ ID NO:

173. rr) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 175, SEQ ID NO: 176, and SEQ ID NO:

177. ss) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 179, SEQ ID NO: 180, and SEQ ID NO:

181. tt) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 183, SEQ ID NO: 184, and SEQ ID NO:

185. uu) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 187, SEQ ID NO: 188, and SEQ ID NO:

189. vv) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 191, SEQ ID NO: 192, and SEQ ID NO:

193. ww) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 195, SEQ ID NO: 196, and SEQ ID NO:

197. xx) Heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), respectively, having the amino acid sequences shown in SEQ ID NO: 199, SEQ ID NO: 200, and SEQ ID NO:

201. yy) Each having the amino acid sequence shown in SEQ ID NO: 203, SEQ ID NO: 204, and SEQ ID NO: 205, respectively, heavy chain CDR1 (HCDR1), heavy chain CDR2 (HCDR2), and heavy chain CDR3 (HCDR3), or Each of the following heavy chains includes a variable region comprising 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. The binding domain comprises a light chain variable region including light chain CDR1 (LCDR1), light chain CDR2 (LCDR2), and light chain CDR3 (LCDR3), each having the amino acid sequences shown in SEQ ID NO: 219, SEQ ID NO: 220, and SEQ ID NO: 221, respectively.

2. The binding domain includes a heavy chain variable region having an amino acid sequence represented by any one of SEQ ID NOs: 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 at least 80%, 85%, 90%, or 95% sequence identity to those sequences. The binding domain according to claim 1, wherein the binding domain has the amino acid sequence shown in Sequence ID No. 218, or includes a light chain variable region having at least 80%, 85%, 90%, or 95% sequence identity thereto.

3. The binding domain includes a heavy chain variable region having an amino acid sequence represented by any one of SEQ ID NOs: 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. The binding domain according to claim 1, wherein the binding domain includes a light chain variable region having the amino acid sequence shown in Sequence ID No.

218.

4. The binding domain according to claim 1, wherein the binding domain is Fab.

5. A binding portion comprising two binding domains as described in claim 1.

6. A binding portion comprising two binding domains as described in claim 3.

7. The binding portion according to claim 5, wherein the binding portion is an IgG1 or IgG4 antibody.

8. The binding portion according to claim 6, wherein the binding portion is an IgG1 or IgG4 antibody.

9. The binding portion according to claim 7, wherein the binding of the binding portion to the Fc receptor is eliminated or reduced, or the binding of the binding portion to the Fc receptor is promoted.

10. The binding portion according to claim 7, wherein the binding portion exhibits a higher binding signal than a reference antibody comprising two heavy chains having the amino acid sequence shown in SEQ ID NO: 210 and two light chains having the amino acid sequence shown in SEQ ID NO:

211.

11. A pharmaceutical composition comprising an effective amount of a binding domain according to any one of claims 1 to 4, or a binding portion according to any one of claims 5 to 10, and a pharmaceutically acceptable carrier.

12. A pharmaceutical composition comprising an effective amount of a binding domain according to any one of claims 1 to 4 or a binding portion according to any one of claims 5 to 10, and a pharmaceutically acceptable carrier, for use in therapy.

13. A pharmaceutical composition comprising an effective amount of a binding domain according to any one of claims 1 to 4 or a binding portion according to any one of claims 5 to 10, and a pharmaceutically acceptable carrier, for use in the treatment of cancer.

14. One or more nucleic acids comprising a nucleic acid sequence encoding a heavy chain variable region as defined in claim 1 and a nucleic acid sequence encoding a light chain variable region as defined in claim 1.

15. One or more nucleic acids comprising a nucleic acid sequence encoding a heavy chain variable region as defined in claim 3 and a nucleic acid sequence encoding a light chain variable region as defined in claim 3.

16. The nucleic acid sequence encoding the heavy chain variable region further comprises a nucleic acid sequence encoding the CH1 region, according to claim 14.

17. The one or more nucleic acids according to claim 15, wherein the nucleic acid sequence encoding the heavy chain variable region further comprises a CH1 region and nucleic acid sequences encoding the hinge, CH2 and CH3 regions.

18. A cell comprising a nucleic acid sequence encoding a heavy chain variable region as defined in Claim 1 and at least one nucleic acid sequence encoding a light chain variable region as defined in Claim 1.

19. A cell comprising a nucleic acid sequence encoding a heavy chain variable region as defined in claim 3 and at least one nucleic acid sequence encoding a light chain variable region as defined in claim 3.

20. The cell according to claim 18, wherein the nucleic acid sequence encoding the light chain variable region further comprises a CL region.

21. The cell according to claim 19, wherein the nucleic acid sequence encoding the light chain variable region further comprises a CL region.

22. A cell that produces a binding domain according to any one of claims 1 to 4, or a binding portion according to any one of claims 5 to 10.

23. A kit comprising a container having a binding domain according to any one of claims 1 to 4, or a binding portion according to any one of claims 5 to 10.

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