Anti-hepsin antibodies and uses thereof

Abstract

The present application discloses methods of using anti-hepsin antibodies to treat, detect, and diagnose a cancer, and methods of using anti-hepsin pharmaceutical compositions of anti-hepsin antibodies to treat, detect, and diagnose a cancer.

Description

ANTI-HEPSIN ANTIBODIES AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This international patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 658,551, filed June 11, 2024, the entire disclosure of which is incorporated herein by reference.INCORPORATION OF SEQUENE LISTING XML

[0002] A computer readable form of the Sequence Listing XML containing the file named "EpitopeWO Sequence Listing.xml," which is 123,696 bytes in size (as measured in MICROSOFT WINDOWS® EXPLORER) and was created on 11, 2025, is provided herein and is herein incorporated by reference. This Sequence Listing consists of SEQ ID NOs: 1- 136.BACKGROUND OF THE INVENTION

[0003] Cancer is one of the leading causes of death in the developed world and over 14 million new cases of cancer occur globally each year. Genetic and environmental factors can cause cancer and the risk of cancer increases significantly with age. Rates of cancer occurrences are increasing as people live longer and as lifestyle changes occur in the developing world. There is a need for new methods of treating, detecting, and diagnosing cancers.SUMMARY OF THE INVENTION

[0004] Hepsin, a type II transmembrane serine protease, is commonly overexpressed in a variety of epithelial cancers, where its overexpression and concurrent proteolytic activity correlate with tumor progression. Recent studies indicate that hepsin undergoes activation via autocatalysis and subsequent ectodomain shedding, thus highlighting its potential as a serumbased biomarker. Using transgenic mouse models of cancer, hepsin overexpression has previously been implicated in disease progression and, notably, metastases that are neuroendocrine in nature. There is a need for methods of treating, detecting, and diagnosing cancer, inclusive of metastatic variants thereof, comprising targeting and interrogation of hepsin expression.

[0005] In some embodiments, provided herein is a method of treating cancer in a subject comprising administering to the subject an antibody that selectively binds to the non-catalyticchain of the extracellular region of hepsin. In some embodiments, the antibody binds to an epitope on the non-catalytic chain of the extracellular region of hepsin. In some embodiments, the hepsin amino acid sequence comprises SEQ ID NO: 41 or SEQ ID NO: 44. In some embodiments, the non-catalytic chain comprises amino acids 1-90 of SEQ ID NO: 44. In some embodiments, the non-catalytic chain comprises a scavenger receptor cysteine-rich (SRCR) domain.

[0006] In some embodiments, provided herein is a method of treating cancer in a subject comprising administering an antibody that selectively binds to hepsin, wherein the antibody binds to one or more amino acids in one or more of the amino acid sequences: YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and / or RLLEVIS (SEQ ID NO: 218). In some embodiments, the antibody binds to one or more amino acids in each of the amino acid sequences YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and RLLEVIS (SEQ ID NO: 218). In some embodiments, the antibody binds to one or more amino acids in one or more of the amino acid sequences 3-9, 19-29, 51-58, and / or 81-87 of SEQ ID NO: 44. In some embodiments, the antibody binds to one or more amino acids in each of the amino acid sequences 3-9, 19-29, 51 -58, and 81 -87 of SEQ ID NO: 44. In some embodiments, the antibody binds to one or more of amino acids 3, 8, 9, 19, 23, 29, 51, 53, 58, 81 or 87 of SEQ ID NO: 44. In some embodiments, the antibody binds to each of the amino acids 3, 8, 9, 19, 23, 29, 51, 53, 58, 81 and 87 of SEQ ID NO: 44.

[0007] In some embodiments, provided herein is a method of treating cancer comprising administering an antibody that selectively binds to hepsin, wherein the antibody binding to the non-catalytic chain of the extracellular region of hepsin is characterized by cross-linking, proteolysis, liquid chromatography, and mass spectrometry (e.g., as described in Examples 2- 4).

[0008] In some embodiments, provided herein is a method of treating cancer comprising administering an antibody that selectively binds to hepsin, wherein wherein the antibody is linked to one or more payloads. In some embodiments, the one or more payloads comprise a protein degrader (e.g. , a lysosome-targeting receptor binder or a ligand for E3 ubiquitin ligase). In some embodiments, the protein degrader is a degrader for non-cytosolic proteins (e.g., a lysosome-targeting receptor binder). In some embodiments, the protein degrader is a degrader for cytosolic proteins (e.g., an E3 ubiquitin ligase). In some embodiments, the one or more payloads comprise a protein degrader and a member selected from the group consisting of a polyethylene glycol (PEG), an antibiotic, an immune-modulating compound, a peptide tag, anazide, a bacterial toxin, an antibody fragment, and a bactericidal macrocyclic peptide. In some embodiments, the one or more payloads comprise a protein degrader and a member selected from the group consisting of a polyethylene glycol (PEG), an antibiotic, an immune- modulating compound, a peptide tag, an azide, a bacterial toxin, an antibody fragment, a bactericidal macrocyclic peptide.

[0009] In some embodiments, provided herein is a method of treating cancer comprising administering an antibody that selectively binds to hepsin, wherein the hepsin does not selectively bind to serine proteases Matripase, KLK6, KLK7, and KLK.8. In some embodiments, the hepsin is human hepsin. In some embodiments, the hepsin is circulating hepsin. In some embodiments, the hepsin is biologically-active. In some embodiments, the hepsin is biologically-active circulating hepsin.

[0010] In some embodiments, provided herein is a method of treating cancer comprising administering an antibody, wherein the antibody comprises a variable heavy chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9. In some embodiments, the antibody comprises a variable light chain complementarity-determining region CDR-L1, CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20. CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10. In some embodiments, the antibody competes for binding on the non-catalytic chain of the extracellular region of hepsin with an antibody comprising a variable heavy’ chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9. In some embodiments, the antibody competes for binding on the non- catalytic chain of the extracellular region of hepsin with an antibody comprising a variable light chain complementarity-determining region CDR-L1, CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

[0011] In some embodiments, provided herein is a method of treating cancer comprising administering an antibody that selectively binds to hepsin, wherein treating cancer comprises inhibiting or reducing cancer metastasis. In some embodiments, the cancer is metastatic cancer. In some embodiments, the antibody binds to the non-catalytic chain of the extracellular region of hepsin and inhibits hepsin. In some embodiments, the antibody binds to the non-catalytic chain of the extracellular region of hepsin and inhibits the catalytic activity of hepsin. In some embodiments, the cancer is breast cancer, ovarian cancer, prostate cancer, a carcinoma, or a combination thereof

[0012] In some embodiments, provided herein is a method of treating cancer in a subject comprising administering to the subject a composition, the composition comprising an excipient and an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin.

[0013] In some embodiments, provided herein is a method of treating cancer in a subject comprising administering to the subject an antibody comprising: (i) a variable heavy chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9, and (ii) a variable light chain complementarity-determining region CDR-L1, CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR- L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR- L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

[0014] In some embodiments, provided herein is a method of detecting cancer in a subject comprising: obtaining a biological sample from the subject; contacting the biological sample with an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin; determining a level of hepsin present in the biological sample; and based at least in part on the level of hepsin, detecting cancer in a subject.

[0015] In some embodiments, provided herein is a method of diagnosing cancer in a subj ect comprising: obtaining a biological sample from the subject; contacting the biological sample with an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin; determining a level of hepsin present in the biological sample; based at least in part on the level of hepsin, diagnosing the subject with a cancer.INCORPORATION BY REFERENCE

[0016] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0017] The sequences described throughout the application are herein incorporated by reference.BRIEF DISCLOSURE OF THE DRAWINGS

[0018] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings of which:FIG. 1 provides a high-mass matrix-assisted laser desorption / ionization imaging mass spectrometry (MALDI MS) analysis of the control and cross-linked samples of the hepsin protein showing the integrity and aggregation level of the samples;

[0019] FIG. 2 provides a MALDI MS analysis of the control and cross-linked samples of the H5 antibody showing the integrity and aggregation level of the samples;

[0020] FIG. 3 provides a MALDI MS analysis of the control and cross-linked samples for the Hepsin / H5 protein complexes showing detection of one non-covalent protein complex that corresponds to Hepsin / H5;

[0021] FIG. 4 shows an overlap mapping of the hepsin peptides identified after hepsin was submitted to proteolysis by trypsin, chymotrypsin, elastase and thermolysin;

[0022] FIG. 5A shows the nanoflow liquid chromatography (nLC) chromatogram (top) and the sum total of the ions detected by mass spectrometry (bottom) for a trypsin proteolysis of hepsin protein;

[0023] FIG. 5B provides an illustration of the molecular interface between the H5 antibody and the hepsin protein;

[0024] FIG. 6 provides a crystal structure of the extracellular region of the hepsin protein with dark shading on the epitope site; and

[0025] FIG. 7 provides a ribbon structure of the hepsin protein with dark shading on the epitope site and with hepsin amino acids that crosslink to the H5 antibody labeled.

[0026] FIG. 8 provides a ribbon structure of the hepsin protein with dark shading on the epitope site and with hepsin some amino acids that bind with the H5 antibody labeled (Y3, S8, S9, K.19, T23, R58, R8 L and S87).DETAILED DESCRIPTION OF THE INVENTION

[0027] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology7, cell biology7, biochemistry7and immunology7, which are within the skill of the art. Such techniques are explained fully in the literature, such as, Molecular Cloning: A Laboratory Manual, second edition (Sambrook et al., 1989) Cold Spring Harbor Press; Oligonucleotide Synthesis (M. J. Gait, ed. , 1984); Methods in Molecular Biology, Humana Press; Cell Biology7: A Laboratory7Notebook (J. E. Cellis, ed., 1998) Academic Press; Animal Cell Culture (R. I. Freshney, ed., 1987); Introduction to Cell and Tissue Culture (J. P. Mather and P. E. Roberts, 1998) Plenum Press: Cell and Tissue Culture: Laboratory Procedures (A. Doyle, J. B. Griffiths, and D. G. Newell, eds., 1993-1998) J. Wiley and Sons; Methods in Enzymology (Academic Press, Inc.); Handbook of Experimental Immunology7(D. M. Weir and C. C. Blackwell, eds. . Gene Transfer Vectors for Mammalian Cells (J. M. Miller and M. P. Cabs, eds., 1987); Current Protocols in Molecular Biology (F. M. Ausubel et al., eds.. 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds.. 1994); Current Protocols in Immunology (J. E. Coligan et al., eds., 1991); Short Protocols in Molecular Biology (Wiley and Sons, 1999); Immunobiology7(C. A. Janeway and P. Travers, 1997); Antibodies (P. Finch, 1997); Antibodies: a practical approach (D. Catty ., ed., IRL Press, 1988-1989): Monoclonal antibodies: a practical approach (P. Shepherd and C. Dean. eds.. Oxford University Press, 2000); Using antibodies: a laboratory manual (E. Harlow and D. Lane (Cold Spring Harbor Laboratory7Press, 1999); and The Antibodies (M. Zanetti and J. D. Capra, eds., Harwood Academic Publishers, 1995).

[0028] The term " about" includes equal to. and a range that takes into account experimental error in a given measurement and can refer to plus or minus 5, 4, 3, 2 or 1% or anywhere inbetween.

[0029] As used herein, "substantially pure", "isolated" or "purified" refers to material which is at least 50% pure (i.e., free from contaminants), more preferably at least 90% pure, more preferably at least 95% pure, more preferably at least 98% pure, more preferably at least 99% pure. Antibodies can be isolated and purified from the culture supernatant or ascitesmentioned above by saturated ammonium sulfate precipitation, euglobulin precipitation method, caproic acid method, caprylic acid method, ion exchange chromatography (DEAE or DE52), or affinity chromatography using anti-Ig column or a protein A, G or L column using art-recognized conventional methods.

[0030] The terms "polypeptide", "oligopeptide", "peptide" and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by nonamino acids. The terms also encompass an amino acid polymer that has been modified naturally or by intervention; for example, disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification, such as conjugation with a labeling component. Also included within the definition are, for example, polypeptides containing one or more analogs of an amino acid (including, for example, unnatural amino acids, etc.), as well as other modifications known in the art. It is understood that, because the polypeptides of this invention are based upon an antibody, the polypeptides can occur as single chains or associated chains.

[0031] "Polynucleotide," or "nucleic acid," as used interchangeably herein, refer to polymers of nucleotides of any length, and include DNA and RNA. The nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a polymer by DNA or RNA polymerase. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and their analogs. If present, modification to the nucleotide structure may be imparted before or after assembly of the polymer. The sequence of nucleotides may be interrupted by non-nucleotide components. A polynucleotide may be further modified after polymerization, such as by conjugation with a labeling component. Other types of modifications include, for example, "caps", substitution of one or more of the naturally occurring nucleotides with an analog, intemucleotide modifications such as, for example, those with uncharged linkages (e.g. , methyl phosphonates, phosphotriesters, phosphoami dates, carbamates, etc. ) and with charged linkages (e.g., phosphorothioates, phosphorodithioates, etc.), those containing pendant moieties, such as, for example, proteins (e.g., nucleases, toxins, antibodies, signal peptides. ply-L-lysine. etc.), those with intercalators (e.g., acridine, psoralen, etc.), those containing chelators (e.g., metals, radioactive metals, boron, oxidative metals, etc.), those containing alkylators, those with modified linkages (e.g., alpha anomeric nucleic acids, etc.), as well as unmodified forms of the polynucleotide(s). Further, any of the hydroxyl groups ordinarily present in the sugars may be replaced, for example, by phosphonate groups, phosphate groups, protected by standardprotecting groups, or activated to prepare additional linkages to additional nucleotides, or may be conjugated to solid supports. The 5' and 3' terminal OH can be phosphorylated or substituted with amines or organic capping group moieties of from 1 to 20 carbon atoms. Other hydroxyls may also be derivatized to standard protecting groups. Polynucleotides can also contain analogous forms of ribose or deoxyribose sugars that are generally known in the art, including, for example, 2'-O-methyl-, 2'-O-allyl, 2'-fluoro-or 2'-azido-ribose, carbocyclic sugar analogs, .alpha. -anomeric sugars, epimeric sugars such as arabinose, xyloses or lyxoses, pyranose sugars, furanose sugars, sedoheptuloses, acyclic analogs and abasic nucleoside analogs such as methyl riboside. One or more phosphodiester linkages may be replaced by alternative linking groups. These alternative linking groups include, but are not limited to, embodiments wherein phosphate is replaced by P(O)S("thioate"), P(S)S ("dithioate"), (O)NR2 ("amidate"), P(O)R, P(O)OR', CO or CH2 ("formacetal"), in which each R or R' is independently H or substituted or unsubstituted alkyl (1-20 C) optionally containing an ether (-O-) linkage, aryl, alkenyl, cycloalkyl, cycloalkenyl or araldyl. Not all linkages in a polynucleotide need be identical. The preceding description applies to all polynucleotides referred to herein, including RNA and DNA.

[0032] As used herein, the terms "hepsin", "HPN", or TMPRSS 1" refer to a type IT transmembrane serine protease (TTSP) expressed on the surface of epithelial cells. As used herein, hepsin includes all mammalian species of native sequence hepsin, e.g, human, canine, feline, equine, bovine, etc. hepsin (HPN) is one of the most upregulated genes in human prostate cancer and encodes a type-II transmembrane serine protease that is overexpressed in up to 90% of prostate tumors with levels often increased >10 fold. Hepsin is upregulated early in prostate cancer initiation and is maintained at high levels throughout progression and metastasis. In addition, hepsin is also overexpressed in ovarian carcinomas, renal cell carcinomas (e.g, metastatic renal cell carcinomas), and in endometrial cancers. Hepsin overexpression has an important role in the promotion of prostate cancer progression and metastasis. Hepsin may activate pro-urokinase plasminogen activator (pro-uP A) and prohepatocyte growth factor (pro-HGF). Activation of the uPA cell-surface serine protease system and HGF-Met scattering pathway may be responsible for promotion of metastasis by hepsin. The present disclosure provides antibodies that specifically bind to clinically relevant extracellular region of hepsin and that may be used for treatment, detection, and diagnosis of cancer (e.g., metastatic cancer). The hepsin protein is composed of a short N-terminal cytoplasmic domain, a transmembrane domain and a single scavenger receptor cysteine-rich domain that packs tightly against the C-terminal protease domain.

[0033] As used herein, an "anti-hepsin antibody" refers to an antibody that is able to selectively bind to the extracellular portion of hepsin. Provided herein is an isolated or a purified antibody, or antigen-binding fragment thereof, that binds to hepsin that comprises a heavy chain variable region and a light chain variable region. It would be understood that the antibodies described herein can be modified as described below or as known in the art.

[0034] "Antibodies" useful in the present invention encompass, but are not limited to, monoclonal antibodies, polyclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab')2, Fv, Fc, etc.), chimeric antibodies, bispecific antibodies, multispecific antibodies, heteroconjugate antibodies, single chain (ScFv), mutants thereof, fusion proteins comprising an antibody portion (e.g., a domain antibody), humanized antibodies, human antibodies, and any other modified configuration of the immunoglobulin molecule that comprises an antigen recognition site of the required specificity, including glycosylation variants of antibodies, amino acid sequence variants of antibodies, and covalently modified antibodies.

[0035] Depending on the amino acid sequence of the constant domain of its heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD. IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known in the art.

[0036] The "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa or ("K" or "K") and lambda or ("X"), based on the amino acid sequences of their constant domains.

[0037] As used herein, a "monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, i. e. , the individual antibodies comprising the population are identical except for possible naturally-occurring mutations that may be present in minor amounts. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody is directed against a single determinant on the antigen (epitope). The modifier "monoclonal" indicates the character of the antibody as being obtained from a substantially homogeneous population of antibodies and is not to be construed as requiring production of the antibody by any particular method. For example, the monoclonal antibodies to be used in accordance with the present invention may be made by the hybridoma method first described by Kohler and Milstein, 1975. Nature, 256:495. or may be made by recombinant DNA methods such as described in U.S. Pat. No. 4.816,567. The monoclonal antibodies mayalso be isolated from phage libraries generated using the techniques described in McCafferty et al. , 1990, Nature, 348:552-554, for example. Other methods are known in the art and are contemplated for use herein.

[0038] As used herein, a "human antibody" means an antibody having an amino acid sequence corresponding to that of an antibody produced by a human and / or has been made using any of the techniques for making human antibodies known in the art or disclosed herein. This definition of a human antibody includes antibodies comprising at least one human heavy chain polypeptide or at least one human light chain polypeptide. One such example is an antibody comprising murine light chain and human heavy chain polypeptides. Human antibodies can be produced using various techniques known in the art. In one embodiment, the human antibody is selected from a phage library, where that phage library expresses human antibodies (Vaughan etal., 1996, Nature Biotechnology, 14:309-314; Sheets etal., 1998, TWAS' USA, 95:6157-6162; Hoogenboom and Winter, 1991, J. Mol. Biol., 227:381; Marks et al., 1991, J. Mol. Biol., 222:581). Human antibodies can also be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. This approach is described in U.S. Pat. Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016. Alternatively, the human antibody may be prepared by immortalizing human B lymphocytes that produce an antibody directed against a target antigen (such B lymphocytes may be recovered from an individual or may have been immunized in vitro). See, e.g., Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, p. 77 (1985); Boemer et al. , 1991, J. Immunol., 147 (l):86-95; and U.S. Pat. No. 5,750,373.

[0039] A "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy’ chain, either alone or in combination. The variable regions of the heavy and light chain each consist of four framework regions (FR) connected by three complementarity determining regions (CDRs) also known as hypervariable regions. The CDRs in each chain are held together in close proximity7by the FRs and, with the CDRs from the other chain, contribute to the formation of the antigen-binding site of antibodies. There are at least two techniques for determining CDRs: (1) an approach based on cross-species sequence variability (i.e., Kabat et al.. Sequences of Proteins of Immunological Interest, (5th ed., 1991, National Institutes of Health, Bethesda Md.)); and (2) an approach based on crystallographic studies of antigen-antibody complexes (Al-Iazikani et al. (1997) J. Molec. Biol. 273:927-948)). As used herein, a CDR may refer to CDRs defined by either approach or by a combination of both approaches.

[0040] A "constant region" of an antibody refers to the constant region of the antibody light chain or the constant region of the antibody heavy chain, either alone or in combination.

[0041] "Epitope" refers to that portion of an antigen or other macromolecule capable of forming a binding interaction with the variable region binding pocket of an antibody. Such binding interactions can be manifested as an intermolecular contact with one or more amino acid residues of one or more (complementarity-determining regions) CDRs. Antigen binding can involve, for example, a CDR3 or a CDR3 pair or, in some cases, interactions of up to all six CDRs of the VH and VL chains. An epitope can be a linear peptide sequence (z.e., "continuous") or can be composed of noncontiguous amino acid sequences (i.e., "conformational" or "discontinuous"). An antibody can recognize one or more amino acid sequences; therefore, an epitope can define more than one distinct amino acid sequence. Epitopes recognized by antibodies can be determined by peptide mapping and sequence analysis techniques well known to one of skill in the art. Binding interactions are manifested as intermolecular contacts between an epitope on an antigen and one or more amino acid residues of a CDR.

[0042] An epitope that "preferentially binds" or "specifically binds" (used interchangeably herein) to an antibody or a polypeptide is a term well understood in the art, and methods to determine such specific or preferential binding are also well known in the art. An antibody specifically binds or preferentially binds to a target if it binds with greater affinity, avidity, more readily, and / or with greater duration than it binds to other substances. For example, an antibody that specifically or preferentially binds to a hepsin epitope is an antibody that binds this epitope with greater affinity, avidity, more readily, and / or with greater duration than it binds to other hepsin epitopes or non-hepsin epitopes. It is also understood by reading this definition that, for example, an antibody (or moiety or epitope) that specifically or preferentially binds to a first target may or may not specifically or preferentially bind to a second target. As such, "specific binding" or "preferential binding" does not necessarily require (although it can include) exclusive binding. Generally, but not necessarily, reference to binding means preferential binding where the affinity' of the antibody, or antigen-binding fragment thereof, is at least at least 2-fold greater, at least 3-fold greater, at least 4-fold greater, at least 5-fold greater, at least 6-fold greater, at least 7-fold greater, at least 8-fold greater, at least 9- fold greater, 10-fold greater, at least 20-fold greater, at least 30-fold greater, at least 40-fold greater, at least 50-fold greater, at least 60-fold greater, at least 70-fold greater, at least 80-fold greater, at least 90-fold greater, at least 100-fold greater, or at least 1000-fold greater than the affinity of the antibody for unrelated amino acid sequences.

[0043] The terms "hypervariable region" and "CDR" when used herein, refer to the amino acid residues of an antibody which are responsible for antigen-binding. The CDRs comprise amino acid residues from three sequence regions which bind in a complementary manner to an antigen and are known as CDR1, CDR2, and CDR3 for each of the VH and VL chains. In the light chain variable domain, the CDRs ty pically correspond to approximately residues 24-34 (CDRL1), 50-56 (CDRL2) and 89-97 (CDRL3), and in the heavy chain variable domain the CDRs typically correspond to approximately residues 31-35 (CDRH1), 50-65 (CDRH2) and 95-102 (CDRH3) according to Kabat et al. (Id.). It is understood that the CDRs of different antibodies may contain insertions, thus the amino acid numbering may differ. The Kabat numbering system accounts for such insertions with a numbering scheme that utilizes letters attached to specific residues (e.g, 27A, 27B, 27C, 27D, 27E, and 27F of CDRL1 in the light chain) to reflect any insertions in the numberings between different antibodies. Alternatively, in the light chain variable domain, the CDRs ty pically correspond to approximately residues 26-32 (CDRL1), 50-52 (CDRL2) and 91-96 (CDRL3), and in the heavy chain variable domain, the CDRs typically correspond to approximately residues 26-32 (CDRH1), 53-55 (CDRH2) and 96-101 (CDRH3) according to Chothia and Lesk. J. Mol. Biol.. 196: 901-917 (1987)).

[0044] "Identity" refers to sequence similarity7between two peptides or between two nucleic acid molecules. Identity' can each be determined by comparing a position in each sequence which may be aligned for purposes of comparison. When an equivalent position in the compared sequences is occupied by the same base or amino acid, then the molecules are identical at that position; wfien the equivalent site occupied by the same or a similar amino acid residue (e.g., similar in steric and / or electronic nature), then the molecules can be referred to as homologous (similar) at that position. Expression as a percentage of identity refers to a function of the number of identical or similar amino acids at positions shared by the compared sequences. A sequence which is "unrelated" or "non-homologous" shares less than 40% identity7, though preferably' less than 25% identity w ith a sequence of the present invention. In comparing tw o sequences, the absence of residues (amino acids or nucleic acids) or presence of extra residues also decreases the identity.

[0045] As used herein, "identity" means the percentage of identical nucleotide or amino acid residues at corresponding positions in two or more sequences when the sequences are aligned to maximize sequence matching, i.e., taking into account gaps and insertions. Identity can be readily calculated by known methods, including but not limited to those described in Computational Molecular Biology’, Lesk. A. M.. ed., Oxford University Press, New York, 1988; Biocomputing: Informatics and Genome Projects, Smith, D. W., ed.. Academic Press.New York, 1993; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press. New Jersey. 1994; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; and Carillo, H., and Lipman, D., SIAM J. Applied Math., 48: 1073 (1988). Methods to determine identity are designed to give the largest match between the sequences tested. Moreover, methods to determine identity are codified in publicly available computer programs. Computer program methods to determine identity between two sequences include, but are not limited to, the GCG program package (Devereux, J., et al. , Nucleic Acids Research 12(1): 387 (1984)), BLASTP, BLASTN, and FASTA (Altschul, S. F. et al., J. Molec. Biol. 215: 403-410 (1990) and Altschul et al. Nuc. Acids Res. 25: 3389-3402 (1997)). The BLAST X program is publicly available from NCBI and other sources (BLAST Manual, Altschul, S., et al., NCBI NLM NIH Bethesda, Md. 20894; Altschul, S., et al., J. Mol. Biol. 215: 403-410 (1990). The well-known Smith Waterman algorithm may also be used to determine identity.

[0046] Provided herein are neutralizing antibodies or antigen-binding fragments that bind to hepsin and inhibit the activity of hepsin.

[0047] Percentage (%) of inhibition / neutralization by an anti-hepsin antibody or antigenbinding fragment thereof of at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, or greater than negative controls is indicative of a antibody or antigen-binding fragment thereof inhibits or neutralizes hepsin. Percentage of inhibition of hepsin by an anti-hepsin antibody or antigen-binding fragment thereof of less than 2-fold greater than negative controls is indicative of an antibody or antigenbinding fragment thereof that does not inhibit hepsin.

[0048] Antibodies, or antigen-binding fragments thereof, described herein can also be used as immunoconjugates. As used herein, for purposes of the specification and claims, immunoconjugates refer to conjugates comprised of the anti-hepsin antibodies or fragments thereof according to the present invention and at least one therapeutic label. Therapeutic labels include antitumor agents and angiogenesis-inhibitors. Such antitumor agents are known in the art and include, but not limited to, toxins, drugs, enzymes, cytokines, radionuclides, and photodynamic agents. Toxins include, but are not limited to, ricin A chain, mutant Pseudomonas exotoxins, diphtheria toxoid, streptonigrin, boamycin, saporin, gelonin, and pokeweed antiviral protein. Drugs include, but are not limited to, daunorubicin, methotrexate, and calicheamicin. Radionuclides include radiometals. Cytokines include, but are not limitedto, transforming growth factor beta (TGF-(3, interleukins, interferons, and tumor necrosis factors; examples of each of these cytokines and their functions are well known in the art. Photodynamic agents include, but are not limited to, porphyrins and their denvatives. Additional therapeutic labels will be known in the art and are also contemplated herein. The methods for complexing the anti-hepsin mAbs or antigen-binding fragments thereof with at least one agent are well known to those skilled in the art (i.e., antibody conjugates as reviewed by Ghetie et al., 1994, Pharmacol. Ther. 63:209-34). Such methods may utilize one of several available heterobifunctional reagents used for coupling or linking molecules. Linkers for conjugating antibodies to other moi eties are well known in the art and are contemplated herein.

[0049] Methods for conjugating or linking polypeptides are well known in the art. Associations (binding) between antibodies and labels include any means known in the art including, but not limited to, covalent and non-covalent interactions, chemical conjugation as w ell as recombinant techniques.

[0050] Antibodies, or antigen-binding fragments thereof, can be modified using techniques known in the art for various purposes such as, for example, by addition of polyethylene glycol (PEG). PEG modification (PEGylation) can lead to one or more of improved circulation time, improved solubility’, improved resistance to proteolysis, reduced antigenicity and immunogenicity, improved bioavailability, reduced toxicity, improved stability, and easier formulation (for a review, see, Francis et al.. International Journal of Hematology 68:1-18, 1998).

[0051] Other methods of improving the half-life of antibody-based fusion proteins in circulation are also known such as, for example, described in U.S. Patent Nos. 7,091,321 and 6,737,056, each of which is hereby incorporated by reference. Additionally, antibodies and antigen-binding fragments thereof may be produced or expressed so that they do not contain fucose on their complex N-glycoside-linked sugar chains. The removal of the fucose from the complex N-glycoside-linked sugar chains is known to increase effector functions of the antibodies and antigen-binding fragments, including but not limited to, antibody dependent cell-mediated cytotoxicity (ADCC) and complement dependent cytotoxicity (CDC). Similarly, antibodies or antigen-binding fragments thereof that can bind hepsin can be attached at their C-terminal end to all or part of an immunoglobulin heavy chain derived from any antibody isotype, e.g., IgG, IgA, IgE, IgD and IgM and any of the isotype sub-classes, particularly IgGl, IgG2b, IgG2a, IgG3 and IgG4.

[0052] Antibodies, or antigen-binding fragments thereof, that bind to hepsin can also be used for purification of hepsin and / or to detect hepsin levels in a sample or subject.Compositions of antibodies and antigen-binding fragments described herein can be used as non-therapeutic agents (e.g. as affinity purification agents). Generally, in one such embodiment, a protein of interest is immobilized on a solid phase such a Sephadex resin or filter paper, using conventional methods known in the art. The immobilized protein is contacted with a sample containing the target of interest (or fragment thereof) to be purified, and thereafter the support is washed with a suitable solvent that will remove substantially all the material in the sample except the target protein, which is bound to the immobilized antibody. Finally, the support is washed with another suitable solvent, such as glycine buffer, pH 5.0, which will release the target protein.

[0053] An antibody or antigen-binding fragment thereof can be conjugated to, or recombinantly engineered with, an affinity tag (e.g., a purification tag). Affinity tags such as, for example, 6x His tag (His- His- His- His- His- His; SEQ ID NO: 47) are conventional in the art.

[0054] In one aspect, the present disclosure provides for an isolated antibody that selectively binds to circulating hepsin or to the c-terminus of circulating hepsin. An anti-hepsin antibody described herein, in some instances, does not selectively bind to the serine proteases Matripase, KLK6, KLK7, and KLK8. In the exemplary VH and VL sequences provided below, it would be understood that the signal sequence is not to be included in the final variable region sequences.

[0055] In one aspect, an antibody or antigen-binding fragment thereof comprises a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of any one of SEQ ID NOS: 1, 2, and 3. In some embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody comprising that sequence retains the ability to bind to same antigen as of the parent (e.g., cancer associated antigen). In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in the amino acid sequence of any one of SEQ ID NOS: 1, 2, and 3. In some embodiments, substitutions, insertions, or deletions occur in regions outside the CDRs (e.g, in the FRs). Optionally, the antibody comprises the VH sequence of the amino acid sequence of any one of SEQ ID NOS: 1, 2, and 3, including one or more post-translational modifications of that sequence. In a particular embodiment, the VH comprises one, two or three CDRs selected from: (a) CDR-H1 comprising the amino acid sequence of any one of SEQ ID NOS: 16, 18, and 19,(b) CDR-H2 comprising the amino acid sequence of any one of SEQ ID NOS: 11, 13, and 14, and (c) CDR-H3 comprising the amino acid sequence of any one of SEQ ID NOS: 6, 8, and 9.

[0056] In one aspect, an antibody or antigen-binding fragment thereof, is provided, wherein the antibody or antigen-binding fragment thereof comprises a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 4 or 5. In some embodiments, a VL sequence having at least 90%. 91%. 92%. 93%. 94%. 95%. 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody or antigen-binding fragment thereof comprising that sequence retains the ability to bind to same antigen as the parent (e.g., cancer associated antigen). In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in the amino acid sequence of SEQ ID NO: 4 or 5. In some embodiments, the substitutions, insertions, or deletions occur in regions outside the CDRs (<?.g, in the FRs). Optionally, the antibody or antigen-binding fragment thereof comprises the VL sequence of SEQ ID NO: 4 or 5, including post-translational modifications of that sequence. In a particular embodiment, the VL comprises one. two or three CDRs selected from (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 17 or 20; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 12 or 15; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 7 or 10.

[0057] In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a VH as in any of the embodiments provided above, and a VL as in any of the embodiments provided above. In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH and a VL, wherein the VH comprises the amino acid sequence of any one of SEQ ID NOS: 1. 2, and 3. and wherein the VL comprises the amino acid sequence in SEQ ID NO: 4 or 5, and optionally including post- translational modifications of those sequences.

[0058] In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a VH selected from any VH in Table 1. In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a VL selected from any VL in Table 1. In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a VH selected from any VH in Table 1 and a VL selected from any VL in Table 1. In one aspect, an antibody or antigenbinding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereofcomprises a VH selected from any VH in Table 1 and a VL selected from any VL in Table 1, wherein the selected VH and VL are paired according to Table 5. In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a CDR-H3 selected from any CDR-H3 in Table 2 and a CDR-L3 selected from any CDRL3 in Table 2, wherein the selected CDR-H3 and CDRL3 are paired according to Table 5. In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a CDR-H2 selected from any CDR-H2 in Table 2 and a CDR-L2 selected from any CDR-L2 in Table 2, wherein the selected CDR-H2 and CDR-L2 are paired according to Table 5. In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigenbinding fragment thereof comprises a CDR-H1 selected from any CDR-H1 in Table 2 and a CDR-L1 selected from any CDR-L1 in Table 2, wherein the selected CDR-H1 and CDR-L1 are paired according to Table 5. In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a CDR-H1, a CDR-H2, and a CDR-H3 selected from any CDR-H1, a CDR-H2, and a CDR-H3 in Table 2 and a CDR-L1, a CDR-L2, and a CDR-L3 selected from any CDR-L1, CDR-L2, or CDR-L3 in Table 2, wherein the selected CDR-H1 , CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR- L3 are paired according to Table 5. In one aspect, the present disclosure provides an antibody or antigen-binding fragment thereof comprising at least one or both variable regions selected from (a) VH comprising the amino acid sequence of SEQ ID NO: 3 and (b) VL comprising the amino acid sequence of SEQ ID NO: 5.

[0059] In one aspect, the present disclosure provides an antibody or antigen-binding fragment thereof comprising at least one, two, three, four, five, or six CDRs selected from (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 19; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 14; (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 9; (d) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20; (e) CDR- L2 comprising the amino acid sequence of SEQ ID NO: 15; and (f) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 10.

[0060] In one aspect, the present disclosure provides an antibody or antigen-binding fragment thereof comprising at least one, at least two, or all three VH CDR sequences selected from (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 19; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 14; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO:9; and (d) a VL comprising the amino acid sequence of SEQ ID NO: 5.

[0061] In one aspect, the present disclosure provides an antibody or antigen-binding fragment thereof comprising at least one, at least two, or all three VL CDR sequences selected from (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 15; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 10; and a VH comprising the amino acid sequence of SEQ ID NO: 3.

[0062] In one aspect, the present disclosure provides an antibody or antigen-binding fragment thereof comprising the CDRs: CDR-H3 comprising the amino acid sequence of SEQ ID NO: 9; and CDR-L3 comprising the amino acid sequence of SEQ ID NO: 10.

[0063] In one aspect, the disclosure herein provides an antibody or antigen-binding fragment thereof comprising at least one, at least two, or all three VL CDR sequences selected from (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 15; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 10. In one aspect, the disclosure provides an antibody or antigenbinding fragment thereof comprising at least one, at least two, or all three VH CDR sequences selected from (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 19; (b) CDR- H2 comprising the amino acid sequence of SEQ ID NO: 14; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 9.

[0064] In one aspect, the disclosure provides an antibody or antigen-binding fragment thereof comprising the CDRs: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 19; (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 14; (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 9; (d) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20; (e) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 15; and (f) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 10.

[0065] In one aspect, an antibody or antigen-binding fragment thereof comprises a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 3. In some embodiments, a VH sequence having at least 90%, 91%, 92%, 93%, 94%, 95%. 96%, 97%, 98%, or 99% identity contains substitutions (e.g, conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody or antigen-binding fragment thereof comprising that sequence retains the ability to bind to antigen. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in the amino acid sequence of SEQ ID NO: 3. In some embodiments, substitutions, insertions, or deletions occur in regions outside the CDRs (e.g, in the FRs). Optionally, the antibody or antigen-binding fragment thereofcomprises the VH sequence of the amino acid sequence of SEQ ID NO: 3, including post- translational modifications of that sequence. In a particular embodiment, the VH comprises one, two or three CDRs selected from: (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 19, (b) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 14, and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 9.

[0066] In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody comprises a light chain variable domain (VL) having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, a VL sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity contains substitutions (e.g., conservative substitutions), insertions, or deletions relative to the reference sequence, but an antibody or antigen-binding fragment thereof comprising that sequence retains the ability to bind to antigen. In some embodiments, a total of 1 to 10 amino acids have been substituted, inserted and / or deleted in the amino acid sequence of SEQ ID NO: 5. In some embodiments, the substitutions, insertions, or deletions occur in regions outside the CDRs (e.g., in the FRs). Optionally, the antibody or antigen-binding fragment thereof comprises the VL sequence of SEQ ID NO: 5, including post-translational modifications of that sequence. In a particular embodiment, the VL comprises one, two or three CDRs selected from (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 20; (b) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 15; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 10.

[0067] In one aspect, an antibody or antigen-binding fragment thereof is provided, wherein the antibody or antigen-binding fragment thereof comprises a VH as in any of the embodiments provided above, and a VL as in any of the embodiments provided above. In some embodiments, the antibody or antigen-binding fragment thereof comprises a VH comprising the amino acid sequence of SEQ ID NO: 3, and a VL sequence in SEQ ID NO: 5, including post-translational modifications of those sequences.

[0068] Hepsin proteins can be composed of a short N-terminal cytoplasmic domain, a transmembrane domain and a single scavenger receptor cysteine-rich (SRCR) domain that packs tightly against the C-terminal protease domain.

[0069] The present disclosure provides antibodies that bind to hepsin and are useful for detecting, diagnosing, and treating cancer. In some embodiments, antibodies described herein bind to and inhibit, inactivate, and / or degrade hepsin. In some embodiments, antibodies described herein bind to and label hepsin. such as for diagnosis (e.g, of cancer) and / or detection (e.g.. of cancer). In some embodiments, antibodies descnbed herein bind to. label(e.g, for diagnostics or detection), and inhibit, inactivate, and / or degrade hepsin. In some embodiments, antibodies provided herein specifically bind to an epitope on the non-catalytic chain of the extracellular region of hepsin and may be used for treatment, detection, and diagnosis of cancer (e g., metastatic cancer). In some embodiments, antibodies provided herein specifically bind to an epitope on the non-catalytic chain that comprises a scavenger receptor cysteine-rich (SRCR) domain. In some embodiments, antibodies provided herein specifically bind to the non-catalytic chain of hepsin and inhibit hepsin (e.g.. inhibit the catalytic activity of hepsin). In some embodiments, antibodies provided herein specifically bind to the non- catalytic chain of hepsin and inhibit the catalytic activity of hepsin (e.g., by inhibiting the proteolytic activity of hepsin).

[0070] In some embodiments, The hepsin described herein, in some instances, comprises human hepsin. In some embodiments, the hepsin in biologically-active extracellular hepsin. In some embodiments, the biologically-active extracellular hepsin further comprises a thrombin cleavage site and, optionally, a spacer. In some embodiments, the hepsin is circulating. In some embodiments, the circulating hepsin comprises an amino acid sequence of SEQ ID NO: 44. In some embodiments, the human hepsin comprises wild-type human hepsin. In some embodiments, the wild-type human hepsin comprises an amino acid sequence of SEQ ID NO: 41.

[0071] In another aspect, the present disclosure provides for an isolated antibody that binds to a recombinant hepsin sequence that comprises an amino acid sequence of SEQ ID NO: 43. In some instances, the binding is selective.

[0072] In some embodiments, the antibody binding to the non-catalytic chain of the extracellular region of hepsin is characterized by cross-linking, proteolysis, liquid chromatography, and mass spectrometry’. The antibody binding to the non-catalytic chain of the extracellular region of hepsin can be characterized using one or more of cross-link, proteolysis, liquid chromatography, and mass spectrometry techniques.

[0073] In some embodiments, antibodies described herein are useful for treating cancers described herein. In some embodiments, a method of treating cancer provided herein comprises administering to a subject an antibody that selectively binds to the non-catalytic chain (e.g., an SRCR domain) of the extracellular region of a hepsin protein. In some embodiments, a method of treating cancer provided herein comprises administering to a subject an antibody that selectively binds to an epitope on the non-catalytic chain (e.g., an SRCR domain) of the extracellular region of a hepsin protein. In some embodiments, a method of treating cancer provided herein comprises administering to a subject an antibody that binds (e.g., selectively)to the non-catalytic chain (e.g, an SRCR domain) of the extracellular region of a protein described herein (e.g, hepsin), such as a hepsin protein comprising the amino acid sequence of SEQ ID NO: 41. In some embodiments, a method of treating cancer provided herein comprises administering to a subject an antibody that binds (e.g, selectively) to the non- catalytic chain (e.g, an SRCR domain) of the extracellular region of a protein (e.g, hepsin), such as a hepsin protein comprising the amino acid sequence of SEQ ID NO: 44. In some embodiments, the non-catalytic chain (e.g, the SRCR domain) of the extracellular region of a hepsin protein comprises amino acids 1-90 of SEQ ID NO: 44.

[0074] In some embodiments of a method provided herein, an antibody (e.g, an antibody provided herein) binds to one or more amino acids in one or more of the amino acid sequences: YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and / or RLLEVIS (SEQ ID NO: 218). In some embodiments, an antibody provided herein binds to one or more amino acids in the amino acid sequence YPVQVSS (SEQ ID NO: 215). In some embodiments, an antibody provided herein binds to one or more amino acids in the amino acid sequence KTEGTWRLLCS (SEQ ID NO: 216). In some embodiments, an antibody provided herein binds to one or more amino acids in the amino acid sequence THSELDVR (SEQ ID NO: 217). In some embodiments, an antibody provided herein binds to one or more amino acids in the amino acid sequence RLLEVIS (SEQ ID NO: 218). In some embodiments of a method provided herein, an antibody (e.g, an antibody provided herein) binds to one or more amino acids in each of the amino acid sequences: YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and RLLEVIS (SEQ ID NO: 218).

[0075] In some embodiments provided herein, an antibody (e.g, an antibody provided herein) selectively binds to an epitope of a protein (e.g, hepsin), the epitope comprising one or more amino acid sequences selected from the group consisting of YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and RLLEVIS (SEQ ID NO: 218). In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acid sequence YPVQVSS (SEQ ID NO: 215). In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acid sequence KTEGTWRLLCS (SEQ ID NO: 216). In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acid sequence THSELDVR (SEQ ID NO: 217). In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acid sequence RLLEVIS (SEQ ID NO: 218). In some embodiments provided herein, an antibody provided herein selectively binds to an epitope of a protein (e.g.hepsin), the epitope comprising amino acid sequences YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and RLLEVIS (SEQ ID NO: 218).

[0076] In some embodiments, an antibody provided herein selectively binds to an epitope having at least 60%, 65%, 70%, 73%, 75%, 77%, 79%, 80%, 83%, 85%, 87%, 89%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 215 or 100% sequence identity to the amino acid sequence of SEQ ID NO: 215. In some embodiments, an antibody provided herein selectively binds to an epitope having at least 60%, 65%, 70%, 73%, 75%, 77%, 79%, 80%, 83%, 85%, 87%, 89%, 90%, 91%, 92%. 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 216 or 100% sequence identity to the amino acid sequence of SEQ ID NO: 216. In some embodiments, an antibody provided herein selectively binds to an epitope having at least 60%, 65%, 70%, 73%, 75%, 77%, 79%, 80%, 83%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity' to the amino acid sequence of SEQ ID NO: 217 or 100% sequence identity to the amino acid sequence of SEQ ID NO: 217. In some embodiments, an antibody provided herein selectively binds to an epitope having at least 60%, 65%, 70%, 73%, 75%, 77%, 79%, 80%, 83%, 85%, 87%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of SEQ ID NO: 218 or 100% sequence identity to the amino acid sequence of SEQ ID NO: 218.

[0077] In some embodiments of a method provided herein, an antibody binds to one or more amino acids in one or more of the amino acid sequences 3-9, 19-29, 51-58, and / or 81-87 of SEQ ID NO: 44. In some embodiments of a method provided herein, an antibody binds to one or more amino acids in one or more of the amino acid sequences 3-9 of SEQ ID NO: 44. In some embodiments of a method provided herein, an antibody binds to one or more amino acids in one or more of the amino acid sequences 19-29 of SEQ ID NO: 44. In some embodiments of a method provided herein, an antibody binds to one or more amino acids in one or more of the amino acid sequences 51-58 of SEQ ID NO: 44. In some embodiments of a method provided herein, an antibody binds to one or more amino acids in one or more of the amino acid sequences 81-87 of SEQ ID NO: 44. In some embodiments of a method provided herein, an antibody binds to one or more amino acids in each of the amino acid sequences 3-9, 19-29, 51-58, and 81-87 of SEQ ID NO: 44.

[0078] In some embodiments provided herein, an antibody (e.g, an antibody provided herein) selectively binds to an epitope of a protein (e.g. hepsin). the epitope comprising one ormore amino acid sequences, the amino acid sequences selected from the group consisting of 3- 9, 19-29, 51-58, and 81-87 of SEQ ID NO: 44. In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acids 3-9 of SEQ ID NO: 44. In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acids 19-29 of SEQ ID NO: 44. In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acids 51-58 of SEQ ID NO: 44. In some embodiments, an antibody provided herein selectively binds to an epitope comprising amino acids 81-87 of SEQ ID NO: 44. In some embodiments provided herein, an antibody provided herein that selectively binds to an epitope of a protein (<?.g, hepsin), the epitope comprising amino acid sequences 3-9, 19-29, 51-58. and 81-87 of SEQ ID NO: 44.

[0079] Provided herein in some embodiments is an antibody that binds to a protein (e.g., hepsin) at one or more amino acids, the amino acids selected from the group consisting of amino acids 3, 8, 9, 19, 23, 29, 51, 53, 58, 81 and 87 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 3 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 8 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 9 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 19 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 23 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 29 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 51 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 53 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 58 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 81 of SEQ ID NO: 44. In some embodiments, an antibody provided herein binds to hepsin at amino acid 87 of SEQ ID NO: 44. In some embodiments, an antibody (e.g, an antibody provided herein) binds to each of the amino acids 3, 8, 9, 19, 23, 29, 51, 53, 58, 81 and 87 of SEQ ID NO: 44.

[0080] The disclosure provides isolated nucleic acids comprising a sequence encoding an antibody polypeptide or antigen-binding fragment thereof. In some embodiments, the isolated nucleic acid comprises a sequence encoding a heavy chain polypeptide of an antibody. See, Tables 3, 4, and 6. In some embodiments, the nucleic acid sequence encoding a heavy chain polypeptide is selected from any one of SEQ ID NOS: 21, 22, and 23. In some embodiments, the isolated nucleic acid comprises a sequence encoding a light chain polypeptide of an antibody. In some embodiments, the nucleic acid sequence encoding a light chain polypeptideis selected from any one of SEQ ID NOS: 24 or 25. In some embodiments, the isolated nucleic acid comprises a sequence encoding a CDR1 polypeptide of a variable heavy chain. In some embodiments, the isolated nucleic acid comprises a sequence encoding a CDR2 polypeptide of a variable heavy chain. In some embodiments, the isolated nucleic acid comprises a sequence encoding a CDR3 polypeptide of a variable heavy chain. In some embodiments, the nucleic acid sequence encoding the CDR1 polypeptide of a variable heavy chain is selected from any one of SEQ ID NOS: 16, 18, and 19. In some embodiments, the nucleic acid sequence encoding the CDR2 polypeptide of a variable heavy chain is selected from any one of SEQ ID NOS: 11, 13, and 14. In some embodiments, the nucleic acid sequence encoding the CDR3 polypeptide of a variable heavy chain is selected from any one of SEQ ID NOS: 6, 8, and 9. In some embodiments, the isolated nucleic acid comprises a sequence encoding a CDR1 polypeptide of a variable light chain. In some embodiments, the isolated nucleic acid comprises a sequence encoding a CDR2 polypeptide of a variable light chain. In some embodiments, the isolated nucleic acid comprises a sequence encoding a CDR3 polypeptide of a variable light chain. In some, embodiments, the nucleic acid sequence encoding the CDR1 region of a variable light chain polypeptide is selected from any one of SEQ ID NOS: 17 and 20. In some, embodiments, the nucleic acid sequence encoding the CDR2 region of a variable light chain polypeptide is selected from any one of SEQ ID NOS: 12 and 15. In some, embodiments, the nucleic acid sequence encoding the CDR3 region of a variable light chain polypeptide is selected from any one of SEQ ID NOS: 7 and 10.

[0081] An antibody described herein can compete for binding on the non-catalytic chain of the extracellular region of hepsin with an antibody comprising a variable heavy chain complementarity -determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9.

[0082] An antibody described herein can compete for binding on the non-catalytic chain of the extracellular region of hepsin with an antibody comprising a variable light chain complementarity-determining region CDR-L1, CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

[0083] An antibody described herein can compete for binding on the non-catalytic chain of the extracellular region of hepsin with an antibody comprising: (i) a vanable heavy chaincomplementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9, and (ii) a variable light chain complementarity-determining region CDR-L1, CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR- L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15. and CDR- L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

[0084] In another aspect, present disclosure provides for a composition that comprises an antibody of any one of the preceding claims and a pharmaceutically acceptable excipient.

[0085] An antibody described herein may be prepared as a lyophilized form or an aqueous solution for storage. A lyophilized antibody may be reconstituted for use by mixing the antibody having the desired degree of purity' with one or more acceptable carriers, excipients, or stabilizers. Acceptable carriers, excipients, or stabilizers are generally nontoxic and may comprise buffers such as phosphate, citrate, and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens, such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosacchandes, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g, Zn-protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG). One or more of such agents may extend the half-life of the antibody in storage.

[0086] Provided herein are compositions of antibodies and antigen-binding fragments thereof that bind hepsin and include those such as described elsewhere herein. Antibodies and antigen-binding fragments thereof that bind hepsin as described herein can be used for the diagnosis of various forms cancer described below.

[0087] Also provided herein are kits for use in the instant methods. Kits may include one or more containers comprising an anti-hepsin antibody described herein and instructions for use in accordance with any of the methods described herein. Generally, these instructionscomprise a description of administration of the anti-hepsin antibody to diagnose a cancer according to any of the methods described herein.

[0088] The containers may be single units or multi units of containers containing an antibody described herein. Instructions supplied in the kits of the invention are typically written instructions on a label or package insert (e.g, a paper sheet included in the kit), but machine- readable instructions (e.g, instructions carried on a magnetic or optical storage disk) are also acceptable.

[0089] The label or package insert indicates that the antibody is used for diagnosing a cancer such as, for example, a prostate cancer, an ovarian cancer, or a carcinoma. In one instance, the cancer is an ovarian cancer. In another instance, the cancer is a prostate cancer. In another instance, the cancer is a carcinoma. Non-limiting examples of carcinomas include, but are not limited to, a renal cell carcinoma (RCC) or a metastatic renal cell carcinoma. Instructions may be provided for practicing any of the methods described herein.

[0090] The kits may be provided in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g, sealed Mylar or plastic bags), and the like. Kits may optionally provide additional components such as buffers for reconstitution of a lyophilized antibody and interpretive information. Normally, the kit comprises a container and a label or package insert(s) on or associated with the container.

[0091] One aspect of the present disclosure relates to diagnosis of a disorder associated with elevated levels of hepsin in a subject. An "individual" or a "subject" to be diagnosed by a method herein may be a mammal, more preferably a human. Mammals also include, but are not limited to, farm animals, sport animals, and pets, including, but not limited to, primates, equines, bovines, alpacas, dogs, cats, rabbits, mice and rats.

[0092] In one instance, the disorder associated with elevated levels of hepsin is a cancer or a tumor. The terms "cancer" and "tumor" are used herein to refer to a cancerous tissue (as compared to expression by normal tissue of the same type). Tumors can include solid tumors (cancers) and semi-solid tumors (cancers). Tumors (cancers) may also, in some instances, be metastatic. In one instance, a tumor or a cancer is an epithelial cancer. Examples of tumors (cancers) include, but are not limited to, a prostate cancer, an ovarian cancer, or a combination thereof. Other examples of epithelial tumors (cancers) include, but are not limited to, a carcinoma, an adenocarcinoma, or a combination thereof. In one instance, the cancer is an ovarian cancer. In another instance, the cancer is a prostate cancer. In another instance, the cancer is a carcinoma. Non-limiting examples of carcinomas include, but are not limited to, a renal cell carcinoma (RCC) or a metastatic renal cell carcinoma.

[0093] Assessment may be performed based on objective measures such as, for example, testing a biological sample obtained from a subject. Further types of assessments are described below. Assessment may also be performed based on subjective measures, such as characterization of symptoms of a subj ect.

[0094] One aspect of the present disclosure provides for a method of identifying the presence of circulating hepsin in a biological sample, the method comprising: (a) contacting the biological sample (e.g, obtained from an individual, such as an individual suspected of having or at risk for cancer) with an antibody that selectively binds circulating hepsin; and (b) determining whether circulating hepsin is present in the biological sample (e.g, determining whether the amount of circulating hepsin is elevated.

[0095] A "biological sample" as used herein, includes, but is not limited to, any quantity of a substance from a living thing or formerly living things such as, for example, humans, mice, rats, monkeys, dogs, rabbits, and other animals. Such samples include, but are not limited to, blood (or non-tissue) sample (e.g, whole blood, serum, urine, etc.), cells, organs, tissues, and combinations thereof In some instances, a biological sample is treated or modified prior to use in a diagnostic method described herein. For example, heparin may be added to a blood sample and serum collected. If a sample is a tissue sample, fluid around the tissue may be collected for use, or a tissue sample may be homogenized in a buffered solution prior to use in a diagnostic method described herein.

[0096] Any suitable assay may be utilized for the described diagnostic methods including, but not limited to, an enzyme-linked immunosorbent assay (ELISA), ELISPOT, immunohistochemistry (IHC), antibody adaptation to microbeads for multiplex and / or microfluidic platforms.

[0097] In some of such diagnostic methods, the method further comprises identifying presence of or risk of developing cancer in a subject (e.g, the subject suspected of or at risk for cancer). In some of such diagnostic methods, the method further comprises identifying risk of recurrence of cancer in a subject (e.g., a subject suspected of being at risk for recurrence). In some of such diagnostic methods, the method further comprises identifying risk of metastasis of cancer in an individual (e.g , an individual suspected of being at risk for recurrence).

[0098] A cancer to be detected using the diagnostic methods described herein comprises an epithelial cancer. Epithelial cancers to be diagnosed using the methods described herein include, but are not limited to, an ovarian cancer, a prostate cancer, a carcinoma, or a combination thereof In one instance, the cancer is an ovarian cancer. In another instance, theT1cancer is a prostate cancer. In another instance, the cancer is a carcinoma. Non-limiting examples of carcinomas include, but are not limited to, a renal cell carcinoma (RCC) or a metastatic renal cell carcinoma.

[0099] Provided herein in some embodiments is a method of identifying the presence of circulating hepsin in a biological sample. In some embodiments, the method comprises contacting the biological sample with an antibody that binds to hepsin (e.g., selectively binds to an epitope on the non-catalytic chain of the extracellular region of hepsin). In some embodiments, the method comprises determining whether hepsin is present in the biological sample. Determining whether hepsin is present in the biological sample can comprise determining whether the amount of hepsin in the sample is elevated (e.g., above a baseline or a threshold).

[0100] Provided herein some embodiments is a method of detecting cancer in a subject. In some embodiments, the method comprises obtaining a biological sample from the subject. In some embodiments, the method comprises contacting the biological sample with an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin. In some embodiments, the method comprises determining a level of hepsin present in the biological sample. In some embodiments, the method comprises, based at least in part on the level of hepsin, detecting cancer in a subject.

[0101] Provided in some embodiments herein is a method of diagnosing cancer in a subject. In some embodiments, the method comprises contacting a biological sample with an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin. In some embodiments, the method comprises obtaining the biological sample from the subject. In some embodiments, the method comprises determining a level of hepsin present in the biological sample. In some embodiments, the method of diagnosing cancer is based at least in part on the level of hepsin. In some embodiments, a subject has or is suspected of having cancer.

[0102] One aspect of the present disclosure relates to the treatment of a disorder associated with elevated levels of circulating hepsin. An "individual" or a "subject" to be treated by a method herein may be a mammal, more preferably a human. Mammals also include, but are not limited to, farm animals, sport animals, and pets, including, but not limited to, primates, equines, bovines, alpacas, dogs, cats, rabbits, mice, and rats. It will be appreciated that a "subject in need thereof may be suffering from a disorder associated with elevated levels of circulating hepsin.

[0103] Some aspects of the present disclosure relate to the treatment of cancer (e.g., comprising administering to a subject an antibody that binds to hepsin). In some embodiments.antibodies described herein are useful for treating epithelial cancer. In some embodiments, exemplary epithelial cancers to be treated using the methods described herein include, but are not limited to, an ovarian cancer, a prostate cancer, a carcinoma, or a combination thereof. In some embodiments, the cancer is an ovarian cancer. In some embodiments, the cancer is a prostate cancer. In certain embodiments, the cancer is a carcinoma. In certain embodiments, the cancer is a metastatic cancer. Non-limiting examples of carcinomas include, but are not limited to, a renal cell carcinoma (RCC) or a metastatic renal cell carcinoma. In some embodiments, a cancer that can be treated using the treatment methods described herein comprises a metastatic cancer. In some embodiments, treating cancer comprises inhibiting or reducing cancer metastasis.

[0104] As used herein, a "therapeutically effective dosage" or a "therapeutically effective amount" of a pharmaceutical composition described herein is an amount sufficient to result in beneficial or desired results in a subj ect. Beneficial or desired results include results such as lessening the severity or delaying the progression of the disorder, including biochemical, histological and / or behavioral symptoms of the disorder, its complications and intermediate pathological phenotypes presenting during development of the disorder.

[0105] As is understood in the clinical context, an effective dosage of a pharmaceutical composition may or may not be achieved in conjunction with another drug, compound, or pharmaceutical composition. Thus, an "effective dosage" may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable result may be or is achieved. Accordingly, in some instances, one or more therapeutic agents may be administered to the subject. In other instances, treatment with a pharmaceutical composition described herein is conducted prior to, or after, one or more other treatment modalities described herein.

[0106] Various formulations of an anti-hepsin antibody may be used for administration. In some embodiments, the anti-hepsin antibody may be administered neat. In some embodiments, the anti-hepsin antibody and a pharmaceutically acceptable excipient may be in various formulations. Pharmaceutically acceptable excipients are known in the art and include relatively inert substances that facilitate administration of a pharmacologically effective substance. For example, an excipient can give form or consistency, or act as a diluent. Suitable excipients include but are not limited to stabilizing agents, wetting and emulsifying agents, salts for varying osmolarity, encapsulating agents, buffers, and skin penetration enhancers.Excipients as well as formulations for parenteral and non-parenteral drug deliver}' are set forth in Remington, The Science and Practice of Pharmacy 20th Ed. Mack Publishing (2000).

[0107] The antibodies may be formulated for administration by any suitable means. The particular dosage regimen, i .e., dose, timing and repetition, will depend on the particular individual and that individual's medical history'. In some instances, these antibodies are formulated for administration by injection (e.g. , intravenously, intraperitoneally, subcutaneously, intramuscularly, etc. ). Accordingly, these antibodies can be combined with pharmaceutically acceptable vehicles such as saline, Ringer's solution, dextrose solution, and the like. In other instances, the antibodies can also be administered via inhalation.

[0108] Generally , for administration of anti-hepsin antibodies, an initial dosage can be about 2 mg / kg. For the purpose of the present invention, a typical daily dosage can be up to 3 pg / kg, up to about 30 pg / kg, up to about 300 pg / kg, up to about 3 mg / kg, up to 30 mg / kg, up to 100 mg / kg or more, or any integer therebetween, depending on the factors mentioned above. For example, a dosage of about 1 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 10 mg / kg, or about 25 mg / kg may be used. For repeated administrations over several days or longer, depending on the condition, the treatment is sustained until a desired suppression of symptoms occurs or until sufficient therapeutic levels are achieved, for example, to reduce pain. An exemplary dosing regimen comprises administering an initial dose of about 2 mg / kg, followed by a weekly maintenance dose of about 1 mg / kg of the anti-hepsin antibody, or followed by a maintenance dose of about 1 mg / kg every other week.

[0109] However, other dosage regimens may be useful, depending on the pattern of pharmacokinetic decay that the practitioner wishes to achieve. For example, in some embodiments, dosing from one-four times a week is contemplated. The progress of this therapy is easily monitored by conventional techniques and assays. The dosing regimen can vary over time. The appropriate dosage of an anti-hepsin antibody will depend on the anti-hepsin antibody employed, the type and stage of cancer to be treated, previous surgery and / or therapy, the subject's clinical history and response to the antibody, and the discretion of the attending physician. Typically, a clinician will administer an anti-hepsin antibody, until a dosage is reached that achieves the desired result. Dose and / or frequency can vary over course of treatment.

[0110] Empirical considerations, such as the half-life, generally will contribute to the determination of the dosage. For example, antibodies that are compatible with the human immune system, such as humanized antibodies or fully human antibodies, may be used toprolong half-life of the antibody and to prevent the antibody being attacked by the host's immune system.

[0111] Frequency of administration may be determined and adjusted over the course of therapy. Alternatively, sustained continuous release formulations of anti-hepsin antibodies may be appropriate. Various formulations and devices for achieving sustained release are know n in the art. To assess efficacy of an anti-hepsin antibody, an indicator of the disease can be followed.

[0112] Treatment includes, for example, about 2-fold, about 3-fold, about 4-fold, about 5- fold, about 10-fold, about 20-fold, about 50-fold, or any fold reduction in between of one or more symptoms. Similarly, treatment can include about 1%, about 2%, about 3%, about 4%, about 5%, about 10%, about 20%, about 30%. about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 99%, 100%, or any percentage reduction in between or one or more symptoms.

[0113] Provided herein in some embodiments is a method of treating cancer in a subject comprising administering to a subject an antibody that binds (e.g, selectively) to hepsin. An antibody provided herein can bind to the extracellular region of hepsin. For example, an antibody of a method provided herein can bind to the non-catalytic chain of the extracellular region of hepsin. Provided in some embodiments herein is a method of treating cancer in a subject comprising administering to a subject an antibody that selectively binds to the non- catalytic chain of the extracellular region of hepsin.

[0114] In some aspects, a method of treating cancer in a subject (e.g., a subject in need thereof) provided herein comprises administering to the subject an antibody comprising: (i) a variable heavy chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19. CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9, and (ii) a variable light chain complementanty-determining region CDR-L1, CDR-L2 and CDR- L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

[0115] One aspect of the present disclosure relates to a method of treating a disorder associated with elevated levels of circulating hepsin in a subject in need thereof, comprisingadministering to the subject an antibody that selectively binds to circulating hepsin as described herein.

[0116] Another aspect of the present disclosure relates to a method of treating hyperh epsinemia in a subject in need thereof, comprising administering to the subject an antibody that selectively binds to circulating hepsin as described herein.

[0117] Treatment is continued until one or more symptoms of the disorders are partially or completely resolved.

[0118] Provided in some embodiments herein is an antibody (e.g. , an antibody of a method provided herein that can bind to the non-catalytic chain of the extracellular region of hepsin) linked to a payload. An antibody provided herein can be linked to one or more payloads. For example, an antibody can be linked to 2, 3. 4, 5, 6, 7. 8, 9, 10, 15, or 20 payloads. The payloads linked to an antibody can be the same (e.g., of the same type) or different (e.g, of different types). Exemplary payloads include but are not limited to protein degraders, polyethylene glycol (PEG), antibiotics, immune-modulating compounds, peptide tags, azides, bacterial toxins, antibody fragments, and bactericidal macrocyclic peptides.

[0119] In some embodiments provided herein, an antibody provided herein (e.g. an antibody of a method provided herein can bind to the non-catalytic chain of the extracellular region of hepsin) is linked to a protein degrader. Exemplar}' protein degraders include but are not limited to lysosome-targeting receptor binders and ligands for E3 ubiquitin ligase. In some embodiments provided herein, an antibody provided herein can be linked to one or more protein degraders. For example, an antibody can be linked to 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, or 20 protein degraders. In some embodiments provided herein, an antibody can be linked to a protein degrader and a PEG, an antibiotic, an immune-modulating compound, a peptide tag, an azide, a bacterial toxin, an antibody fragment, or a bactericidal macrocyclic peptide.

[0120] In some embodiments provided herein, a protein degrader is a degrader for non- cytosolic (e.g, extracellular or circulating) proteins (e.g., a lysosome-targeting receptor binder). An exemplar ' lysosome-targeting receptor binders is a cation-independent mannose- 6-phosphate receptor (CI-M6PR) receptor binder. In some embodiments provided herein, an antibody provided herein is linked to a lysosome-targeting receptor binder comprising a mannose-6-phosphate receptor (M6PR) moiety.

[0121] In some embodiments provided herein, a protein degrader is a degrader for cytosolic (e.g, intracellular) proteins. For example, a protein degrader provided herein can be an E3 ubiquitin ligase.

[0122] Provided herein in some embodiments is a method of treating cancer in a subject comprising administering to a subject an antibody that binds (e.g. selectively) to hepsin and that is linked to a pay load. Provided herein in some embodiments herein is a method of treating cancer in a subject comprising administering to a subject an antibody that binds (e g., selectively) to hepsin and that is linked to a protein degrader. Provided in some embodiments herein is a method of treating cancer in a subject comprising administering to a subject an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin and that is linked to a protein degrader.

[0123] Provided herein in some embodiments is a method of treating cancer in a subject comprising administering to the subject a composition, the composition comprising an excipient and an antibody that binds to hepsin (e.g. that selectively binds to the non-catalytic chain of the extracellular region of hepsin). Provided herein in some embodiments is a method of treating cancer in a subject comprising administering to the subject a composition, the composition comprising an excipient and an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin.

[0124] Exemplary amino acid and nucleic acid sequences of variable heavy chains, variable light chains, and their corresponding CDR are provided below.

[0125] Table 1 lists exemplary amino acid sequences of variable heavy chain (VH) and exemplary amino acid sequences variable light chain (VL).

[0126] Table 2 below lists exemplary amino acid sequences of complementarity determining regions from variable heavy chains (CDR-H) and exemplary’ amino acid sequences of complementarity determining regions from variable light chains (CDR-L).(VH) and exemplary nucleic acid sequences of variable light chains (VL).determining region from variable heavy chains (CDR-H) and exemplar}7nucleic acid sequences of complementarity determining region from variable light chains (CDR-L). The start and stop position on the corresponding isolated nucleic acid sequence is indicated.

[0130] Table 6 lists exemplary’ nucleic acid sequence heavy’ and light chain pairings.SEQ ID NO: 41: native, wild-type human hepsinMAQKEGGRTVPCCSRPKVAALTAGTLLLLTAIGAASWAIVAVLLRSDQEPLYPVQVSSADARLMVFDKTEGTWRLLCSSRSNARVAGLSCEEMGFLRALTHSELDVRTAGANGTSGFFCVDEGRLPHTQRLLEVISVCDCPRGRFLAAICQDCGRRKLPVDRIVGGRDTSLGRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVLSRWRVFAGAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEYIQPVCLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNGADFYGNQIKPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQKPGVYTKVSDFREWIFQAIKTHSEASGMVTQLSEQ ID NO: 42: Original recombinant hepsin polynucleotide sequence; c-terminal his tag was included for purification purposes.1 cgggacacca gcttgggccg gtggccgtgg caagtcagcc ttcgctatga51 tggagcacac ctctgtgggg gatccctgct ctccggggac tgggtgctga101 cagccgccca ctgcttcccg gagcggaacc gggtcctgtc ccgatggcga151 gtgtttgccg gtgccgtggc ccaggcctct ccccacggtc tgcagctggg201 ggtgcaggct gtggtctacc acgggggcta tcttcccttt cgggacccca251 acagcgagga gaacagcaac gatattgccc tggtccacct ctccagtccc301 ctgcccctca cagaatacat ccagcctgtg tgcctcccag ctgccggcca351 ggccctggtg gatggcaaga tctgtaccgt gacgggctgg ggcaacacgc401 agtactatgg ccaacaggcc ggggtactcc aggaggctcg agtccccata451 atcagcaatg atgtctgcaa tggcgctgac tctatggaa accagatcaa501 gcccaagatg ttctgtgctg gctaccccga gggtggcat gatgcctgcc551 agggcgacag cggtggtccc tttgtgtgtg aggacagcat ctctcggacg601 ccacgtggc ggctgtgtgg cattgtgagt tggggcactg gctgtgccct651 ggcccagaag ccaggcgtct acaccangt cagtgactc cgggagtgga701 tcttccaggc cataaagact cactccgaag ccagcggcat ggtgacccag751 etcSEQ ID NO: 43: Original recombinant hepsin amino acid sequence; c-terminal his tag was included for purification purposes.1 RDTSLGRWPW QVSLRYDGAH LCGGSLLSGD WVLTAAHCFP ERNRVLSRWR51 VFAGAVAQAS PHGLQLGVQA VVYHGGYLPF RDPNSEENSN DIALVHLSSP01 LPLTEYIQPV CLPAAGQALV DGKICTVTGW GNTQYYGQQA GVLQEARVPI51 ISNDVCNGAD FYGNQIKPKM FCAGYPEGGI DACQGDSGGP FVCEDSISRT01 PRWRLCGIVS WGTGCALAQK PGVYTKVSDF REWIFQAIKT HSEASGMVTQ51 LSEQ ID NO: 44: Recombinant hepsin protein with thrombin cleavage site underlined and c-terminal 6x-his-tag in italicsMLYPVQVSSADARLMVFDKTEGTWRLLCSSRSNARVAGLSCEEMGFLRALTHSELD VRTAGANGTSGFFCVDEGRLPHTQRLLEVISVCDCPRGRFLAAICQDCGRRKLPVDRLVPRGSIVGGRDTSLGRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVL SRWRVFAGAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEYIQPVCLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNG ADFYGNQIKPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQKPGVYTKVSDFREWIFQAIKTHSEASGMVTQL7W7W / WSEQ ID NO: 45: Hepsin with FLAG tag underlined and a linker in italicsMAQKEGGRTVPCCSRPKVAALTAGTLLLLTAIGAASWAIVAVLLRSDQEPLYPVQVS S AD ARLMVFDKTEGTWRLLC S SRSNARV AGLSCEEMGFLRALTH SELD VRTAGAN GTSGFFCVDEGRLPHTQRLLEVISVCDCPRGRFLAAICQDCGRRKLPVDRIVGGRDTS LGRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVLSRWRVFAGAVAQ ASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEYIQPVCLPAAG QALVDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNGADFYGNQIKPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQKPGVYTKVS DFREWIFQAIKTHSEASGMVTQLGTGGGXGGGSGGGA.SDYKDDDDKResidues shown in orange represent the inserted linker and residues shown in red represent the Flag tag.SEQ ID NO: 46: Hepsin with thrombin cleavage site underlined, a linker in italics, and a FLAG tag in underlined italicsMAQKEGGRTVPCCSRPKVAALTAGTLLLLTAIGAASWAIVAVLLRSDQEPLYPVQV SSADARLMVFDKTEGTWRLLCSSRSNARVAGLSCEEMGFLRALTHSELDVRTAGAN GTSGFFCVDEGRLPHTQRLLEVISVCDCPRGRFLAAICQDCGRRKLPVDRLyPRGSIV GGRDTSLGRWPWQVSLRYDGAHLCGGSLLSGDWVLTAAHCFPERNRVLSRWRVFA GAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIALVHLSSPLPLTEYIQPV CLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQEARVPIISNDVCNGADFYGNQI KPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCGIVSWGTGCALAQKPG VYTKVSDFREWIFOAIKTHSEASGMVTOLGTGGGSGGGSGGGTXDYKDDDDKResidues shown in green represent the inserted thrombin cleavage site, residues shown in orange represent the inserted linker and residues in red represent the Flag tag.SEQ ID NO: 47: 6x-histidine-tagHis- His- His- His- His- HisSEQ ID NO: 48: Extracellular domain (ECD) with Flag tag, linker and gLUC signal sequence.MGVKVLFALICIAVAEAGRDTSLGRWPWOVSLRYDGAHLCGGSLLSGDWVLTAAH CFPERNRVLSRWRVFAGAVAQASPHGLQLGVQAVVYHGGYLPFRDPNSEENSNDIA LVHLSSPLPLTEYIQPVCLPAAGQALVDGKICTVTGWGNTQYYGQQAGVLQEARVPI ISNDVCNGADFYGNQIKPKMFCAGYPEGGIDACQGDSGGPFVCEDSISRTPRWRLCG IVSWGTGCALAQKPGWTKVSDFREWIFQAIKTHSEASGMVTQLGTGGGSGGGXGGGASDYKDDDDKUnderlined & italicized residues indicate Flag tag, italicized residues indicate inserted linker and underlined residues indicate gLUC signal sequence.EXAMPLES

[0131] The application may be better understood by reference to the following non-limiting examples, which are provided as exemplar}' embodiments of the application. The following examples are presented in order to more fully illustrate embodiments and should in no way be construed, however, as limiting the broad scope of the application.Example 1: Analysis of Hepsin Protein and H5 Antibodies

[0132] A high-mass matrix-assisted laser desorption / ionization imaging mass spectrometry (MALDI MS) analysis of Recombinant Hepsin (SEQ ID NO: 44) and H5 antibody (comprising a reconstructed nucleic acid consensus sequence encoding a heavy chain polypeptide of an antibody comprising SEQ ID NO:23 and a reconstructed nucleic acid consensus sequence encoding a light chain polypeptide of an antibody comprising SEQ ID NO:25) samples was performed to: (i) verify their integrity; (ii) level of multimerization of the hepsin protein and aggregation of the H5 antibody, and (iii) stoichiometry of the antibody / antigen interaction.Materials and MethodsInstrumentation

[0133] The measurements were performed using an Autoflex II MALDI ToF / ToF mass spectrometer (Bruker) equipped with a CovalX HM4 interaction module. The CovalX interaction module contained a special detecting system designed to optimize detection up to 2 MDa with nano-molar sensitivity.Sample Preparation

[0134] The hepsin sample was reconcentrated to reach a concentration of 1.0 mg / ml. Then, 20 pL of each protein sample (hepsin protein and H5 antibody) were pipetted to prepare 8 dilutions with a final volume of 10 pL. These 8 dilutions were prepared to obtain the concentrations detailed in Table 10. After dilution, the samples were prepared for high-mass MALDI MS analysis.Table 10High-Mass MALDI MS Analysis

[0135] High-mass MALDI analysis was performed on control and cross-linked samples.Hepsin ResultsControl Experiments

[0136] In the hepsin control experiments, one main peak was detected for every dilution from 1 to 1 / 32 with an Observed Molecular Weight (MH+) of 42.979 kDA (FIG. 1, top).Cross-Link Experiments

[0137] In the hepsin cross-link experiments, one main peak was detected for every dilution from 1 to 1 / 4 with an Observed Molecular Weight (MH+) of 44.532 kDA (FIG. 1. middle). Using tracker software, no non-covalent complexes were detected in the higher mass range (FIG. 1, bottom).H5 ResultsControl Experiments

[0138] In the H5 control experiments, one main peak was detected for every dilution from 1 to 1 / 32 with an Observed Molecular Weight (MH+) of 149. 130 kDA (FIG. 2, top).Cross-Link Experiments

[0139] In the H5 cross-link experiments, one main peak was detected for every' dilution from 1 to 1 / 16 with an Observed Molecular Weight (MH+) of 158.084 kDA (FIG. 2, middle). Using tracker software, no non-covalent complexes were detected in the higher mass range (FIG. 2, bottom).Conclusion

[0140] Using high-mass MALDI MIMS analysis, there was no detection of signification multimers of hepsin or aggregates of H5.Example 2: Characterization of the H5 / Hepsin Cross-Link Complex

[0141] A high-mass MALDI MS analysis was performed to verify that the intact H5 (comprising a reconstructed nucleic acid consensus sequence encoding a heavy chain polypeptide of an antibody comprising SEQ ID NO: 23 and a reconstructed nucleic acid consensus sequence encoding a light chain polypeptide of an antibody comprising SEQ ID NO:25) I hepsin (SEQ ID NO: 44) cross-link antibody / antigen complex is present in the sample.Materials and MethodsInstrumentation

[0142] The measurements were performed using an Autoflex II MALDI ToF / ToF mass spectrometer (Bruker) equipped with a CovalX HM4 interaction module. The CovalX interaction module contained a special detecting system designed to optimize detection up to 2 MDa with nano-molar sensitivity.Sample Preparation

[0143] A mixture of hepsin and H5 was prepared wi th the concentrations provided in Table11. The mixture was then prepared for high-mass MALDI MS analysis.Table 1 1High-Mass MALDI MS Analysis

[0144] High-mass MALDI analysis was performed on the hepsin and H5 mixture.Hepsin ResultsControl Experiments

[0145] Using high-mass MALDI analysis, hepsin and H5 were detected with an Observed Molecular Weight (MH+) 43.591 kDa and 149.663 kDa, respectively (FIG. 3, top).Cross-Link Experiments

[0146] The cross-linking assay was completed after 180 minutes incubation time with a DSS reagent. After cross-linking, using high-mass MALDI analysis, an additional peak was detected with an Observed Molecular Weight (MH+) of 206.046 kDa (FIG. 3, middle). Using tracker software, the control and cross-link spectra were overlaid. One non-covalent protein complex with an Observed Molecular Mass (MH+) of 193.248 kDA was detected (FIG. 3, bottom).Example 3: Characterization and Peptide Mass Fingerprint of Hepsin

[0147] To characterize hepsin (SEQ ID NO: 44), hepsin was submitted to trypsin, chymotrypsin, elastase and thermolysin proteolysis followed by an nLC-Q-Exactive MS / MS analysis.Materials and MethodsInstrumentation

[0148] For the characterization of hepsin, an nLC Ultimate 3000-RSLC system in line with a Q-Exactive mass spectrometer (Thermo Scientific) was used.Sample Preparation

[0149] Cross-linking, reduction / alkylation and proteolysis with five enzymes (Trypsin, Chymotrypsin. Elastase and Thermolysin) were performed.Liquid Chromatography

[0150] After proteolysis, 1 pl of each peptide solution was loaded onto a nano-liquid chromatography system (Ultimate 3000-RSLC). Liquid chromatography was performed.Mass Spectrometry

[0151] An nLC chromatographic system was in line with the Q-Exactive Plus mass spectrometer. The MS analysis was performed, and data were analyzed.ResultsTrypsin Proteolysis27 peptides were identified in the sequence of hepsin, covering 90.90% of the sequence (Table 12)Table 12Chymotrypsin Proteoly sis

[0152] 46 peptides were identified in the sequence of hepsin, covering 84.75% of the sequence (Table 13).Table 13Elastase Proteolysis

[0153] 29 peptides were identified in the sequence of hepsin, covering 46.25% of the sequence (Table 14).Table 14Thermolysin Proteolysis

[0154] 45 peptides were identified in the sequence of hepsin, covering 60.42% of the sequence (Table 15).Table 15Hepsin Coverage

[0155] Based on the results obtained, overlap mapping of the Trypsin, Chymotry psin, Elastase and Thermolysin peptides was designed (FIG. 4). Combining the peptides of Try psin, Chymotrypsin, Elastase and Thermolysin proteolysis, 99.46% of the sequence is covered.

[0156] The nLC chromatogram and the total sum of the ions detected by the Q-Exactive for the Trypsin digest of hepsin are provided in FIG. 5A and FIG. 5B, respectively.Example 4: Characterization of the Hepsin-H5 Molecular Interfaces

[0157] To determine the epitope of the H5 antibodies (comprising a reconstructed nucleic acid consensus sequence encoding a heavy chain polypeptide of an antibody comprising SEQ ID NO:23 and a reconstructed nucleic acid consensus sequence encoding a light chain polypeptide of an antibody comprising SEQ ID NO:25) on hepsin (SEQ ID NO:44) with high resolution, the protein complexes were incubated with deuterated cross-linkers and subjected to multi-enzy matic cleavage. An individual length of cross-linker was selected which was shown to effectively stabilize the complex using high-mass MALDI detection. A 50:50 mixture of deuterated to undeuterated cross-linker was created to provide a unique mass tag for detecting the linkers' location within the proteins sequence. After enrichment of the crosslinked peptides, the samples were analyzed by high resolution mass spectrometry and the data generated were analyzed using dedicated softwares.Materials and MethodsInstrumentation

[0158] For this analysis, nLC chromatography Ultimate 3000-RSLC (Thermo Scientific) in combination with an Orbitrap Q-Exactive Plus mass spectrometer (Thermo Scientific) were used.Sample Preparation

[0159] A mixture of Hepsin / H5 was prepared with the concentrations provided in Table 16. Cross-linking, reduction / alkylation and proteolysis with five enzymes (Trypsin, Chymotry psin, Elastase and Thermolysin) were performed.Table 16Liquid Chromatography

[0160] After proteolysis, 1 pl of each peptide solution generated by proteolysis w as loaded onto a nano-liquid chromatography system. Liquid chromatography was performed.Mass Spectrometry'

[0161] An nLC chromatographic system was in line with the Q-Exactive Plus mass spectrometer. The MS analysis was performed, and data were analyzed.Results

[0162] After Trypsin, Chymotrypsin, Elastase and Thermolysin proteolysis of the protein complex Hepsin / H5 with deuterated d0dl2, nLC-Q-Exactive MS / MS analysis detected 19 cross-linked peptides between hepsin and H5.

[0163] The sequences and positions of cross-links are provided in Table 17.Table 17

[0164] Using chemical cross-linking, High-Mass MALDI mass spectrometry , and nLC-Q- Exactive mass spectrometry, the molecular interface between H5 and hepsin was characterized. This analysis indicated that the interaction between H5 and hepsin comprises the following ammo acids on hepsin (SEQ ID NO: 44): 3, 8, 9, 19, 23, 29, 51, 53, 58, 81 and 87 (FIG. 6). The analysis further suggested that the epitope comprised one of more of the follow ing amino acid sequences: YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), or RLLEVIS (SEQ ID NO: 218). FIG. 7 provides a Protein Data Bank crystal structure (1Z8G) of the extracellular region of the hepsin protein with dark shading on the epitope sites. Areas with dark shading correspond to the amino acids YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), or RLLEVIS (SEQ ID NO: 218) of the hepsin protein amino acid sequence (SEQ ID NO: 44). FIG. 8 provides a ribbon structure of the hepsin protein with dark shading on the epitope site and with hepsin some amino acids that bind with the H5 antibody labeled (Y3, S8, S9, K19, T23, R58, R81, and S87).Example 5: Method of Treating Cancer

[0165] Testicular cancer will be treated in a subject in need thereof by administering an antibody that binds to the non-catalytic chain of the extracellular region of hepsin. The antibody will comprise (i) a variable heavy chain complementarity-determining region CDR-H1, CDR- H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9, and (ii) a variable light chain complementarity -determining region CDR-L1, CDR- L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20. CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence ofSEQ ID NO: 10. The antibody will bind to one or more of amino acids 3, 8, 9, 19, 23, 29, 51, 53, 58, 81 or 87 of SEQ ID NO: 44.

[0166] Optionally, the antibody will be linked to one or more payloads (for example, one or more protein degraders such as a lysosome-targeting receptor binder).

[0167] While certain embodiments of the present application have been shown and described herein, it will be obvious that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the embodiments; it should be understood that various alternatives to the embodiments described herein may be employed in practicing the methods described herein.

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method of treating cancer in a subject comprising administering to the subject an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin.

2. The method of claim 1 , wherein the antibody binds to an epitope on the non-catalytic chain of the extracellular region of hepsin.

3. The method of any one of the preceding claims, wherein the hepsin amino acid sequence comprises SEQ ID NO: 41 or SEQ ID NO: 44.

4. The method of any one of the preceding claims, wherein the non-catalytic chain comprises amino acids 1-90 of SEQ ID NO: 44.

5. The method of any one of the preceding claims, wherein the non-catalytic chain comprises a scavenger receptor cysteine-rich (SRCR) domain.

6. The method of any one of the preceding claims, wherein the antibody binds to one or more amino acids in one or more of the amino acid sequences: YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and / or RLLEVIS (SEQ ID NO: 218).

7. The method of any one of the preceding claims, wherein the antibody binds to one or more amino acids in each of the amino acid sequences YPVQVSS (SEQ ID NO: 215), KTEGTWRLLCS (SEQ ID NO: 216), THSELDVR (SEQ ID NO: 217), and RLLEVIS (SEQ ID NO: 218).

8. The method of any one of the preceding claims, wherein the antibody binds to one or more amino acids in one or more of the amino acid sequences 3-9, 19-29. 51-58, and / or 81-87 of SEQ ID NO: 44.

9. The method of any one of the preceding claims, wherein the antibody binds to one or more amino acids in each of the amino acid sequences 3-9, 19-29, 51-58, and 81-87 of SEQ ID NO: 44.

10. The method of any one of the preceding claims, wherein the antibody binds to one or more of amino acids 3. 8, 9, 19, 23, 29, 51, 53, 58, 81 or 87 of SEQ ID NO: 44.

11. The method of any one of the preceding claims, wherein the antibody binds to each of the amino acids 3. 8, 9, 19, 23, 29, 51, 53, 58, 81 and 87 of SEQ ID NO: 44.

12. The method of any one of the preceding claims, wherein the antibody binding to the non-catalytic chain of the extracellular region of hepsin is characterized by crosslinking, proteolysis, liquid chromatography, and mass spectrometry (e.g., as described in Examples 2-4).

13. The method of any one of the preceding claims, wherein the antibody is linked to one or more payloads.

14. The method of any one of the preceding claims, wherein the one or more payloads comprise a protein degrader (e.g., a lysosome-targeting receptor binder or a ligand for E3 ubiquitin ligase).

15. The method of any one of the preceding claims, wherein the protein degrader is a degrader for non-cytosolic proteins (e.g., a lysosome-targeting receptor binder).

16. The method of any one of the preceding claims, wherein the protein degrader is a degrader for cytosolic proteins (e.g.. an E3 ubiquitin ligase).

17. The method of any one of the preceding claims, wherein the one or more payloads comprise a protein degrader and a member selected from the group consisting of a polyethylene glycol (PEG), an antibiotic, an immune-modulating compound, a peptide tag, an azide, a bacterial toxin, an antibody fragment, and a bactericidal macrocyclic peptide.

18. The method of any one of the preceding claims, wherein the one or more pay loads comprise a protein degrader and a member selected from the group consisting of a polyethylene glycol (PEG), an antibiotic, an immune-modulating compound, a peptide tag, an azide, a bacterial toxin, an antibody fragment, a bactericidal macrocyclic peptide.

19. The method of any one of the preceding claims, wherein the hepsin does not selectively bind to serine proteases Matripase, KLK6, KLK7, and KLK8.

20. The method of any one of the preceding claims, wherein the hepsin is human hepsin.

21. The method of any one of the preceding claims, wherein the hepsin is circulating hepsin.

22. The method of any one of the preceding claims, wherein the hepsin is biologically - active.

23. The method of any one of the preceding claims, wherein the hepsin is biologically- active circulating hepsin.

24. The method of any one of the preceding claims, wherein the antibody comprises a variable heavy chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9.

25. The method of any one of the preceding claims, wherein the antibody comprises a variable light chain complementarity-determining region CDR-L1. CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

26. The method of anyone of the preceding claims, wherein the antibody competes for binding on the non-catalytic chain of the extracellular region of hepsin with an antibody comprising a variable heavy chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9.

27. The method of anyone of the preceding claims, wherein the antibody competes for binding on the non-catalytic chain of the extracellular region of hepsin with an antibody comprising a variable light chain complementarity-determining region CDR-L1. CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO:

20. CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

28. The method of any one of the preceding claims, wherein treating cancer comprises inhibiting or reducing cancer metastasis.

29. The method of any one of the preceding claims, wherein the cancer is metastatic cancer.

30. The method of any one of the preceding claims, wherein the antibody binds to the non-catalytic chain of the extracellular region of hepsin and inhibits hepsin.

31. The method of any one of the preceding claims, wherein the antibody binds to the non-catalytic chain of the extracellular region of hepsin and inhibits the catalytic activity of hepsin.

32. The method of any one of the preceding claims, wherein the cancer is breast cancer, ovarian cancer, prostate cancer, a carcinoma, or a combination thereof.

33. A method of treating cancer in a subject comprising administering to the subject a composition, the composition comprising an excipient and an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin.

34. A method of treating cancer in a subject comprising administering to the subject an antibody comprising: (i) a variable heavy chain complementarity-determining region CDR-H1, CDR-H2 and CDR-H3, wherein CDR-H1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 19, CDR-H2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 14, and CDR-H3 comprises a reconstructed polypeptide consensus sequence SEQ ID NO: 9, and (ii) a variable light chain complementarity-determining region CDR-L1, CDR-L2 and CDR-L3, wherein CDR-L1 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 20, CDR-L2 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 15, and CDR-L3 comprises a reconstructed polypeptide consensus sequence of SEQ ID NO: 10.

35. A method of detecting cancer in a subject comprising: obtaining a biological sample from the subject: contacting the biological sample with an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin; determining a level of hepsin present in the biological sample; based at least in part on the level of hepsin, detecting cancer in a subject.

36. A method of diagnosing cancer in a subject comprising: obtaining a biological sample from the subject:contacting the biological sample with an antibody that selectively binds to the non-catalytic chain of the extracellular region of hepsin; determining a level of hepsin present in the biological sample; based at least in part on the level of hepsin, diagnosing the subject with a cancer.

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