Monoclonal Antibodies, Compositions and Methods for Detecting Mucin-like Protein (MLP) as a Biomarker for Ovarian and Pancreatic Cancer

Abstract

In various embodiments the invention provides anti-mucin-like protein (MLP) monoclonal antibodies, compositions and methods for detecting MLP as a biomarker for mucin-secreting type of cancer such as ovarian or pancreatic cancer.

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows: 
     
         1 . A method of detecting or diagnosing epithelial cancer by determining the presence or amount of MLP in a biological sample from a test subject, the method comprising:
 (a) contacting a biological sample from a test subject with an anti-MLP antibody or antigen-binding fragment thereof in an in vitro immunoassay; and   (b) detecting the presence or absence of binding of said antibody, wherein the presence of binding indicates the presence or amount of MLP in the sample, wherein the antibody or fragment thereof binds to an epitope in the C-terminal region of MLP, set forth as SEQ ID NO:4.   
     
     
         2 . The method of  claim 1 , wherein said anti-MLP antibody is labeled with a detectable moiety and step (b) comprises detecting the presence or amount of said detectable moiety. 
     
     
         3 . The method of  claim 1 , further comprising comparing the amount of MLP detected in accordance with step (b) with a reference standard or control sample from a healthy subject, wherein an increase of at least two-fold or higher (e.g., at least five-fold, or at least ten-fold) in the level of MLP in the test sample as compared to the control sample (or reference standard) indicates the presence of, or increased risk for developing an epithelial cancer, such as ovarian cancer or pancreatic cancer, in the test subject. 
     
     
         4 . The method of  claim 1 , wherein the biological sample is selected from the group consisting of blood, serum, plasma and tissue. 
     
     
         5 . The method of  claim 1 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody that binds to the same epitope or competes for binding to MLP with a reference antibody selected from the group consisting of:
 the monoclonal anti-MLP antibody Clone 11 produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121699;   the monoclonal anti-MLP antibody Clone B produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121700; and   the monoclonal anti-MLP antibody clone C produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121701.   
     
     
         6 . The method of  claim 1 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody having a heavy chain variable region CDR-H3 sequence comprising an amino acid sequence set forth as SEQ ID NO:15, SEQ ID NO:35, SEQ ID NO:36 or SEQ ID NO:19, and conservative sequence modifications thereof, and having a light chain variable region CDR-L3 sequence comprising an amino acid sequence set forth as SEQ ID NO:23 or SEQ ID NO:27, and conservative sequence modifications thereof. 
     
     
         7 . The method of  claim 1 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody that comprises a heavy chain variable region and/or a light chain variable region set forth in Table 1, and conservative sequence modifications thereof. 
     
     
         8 . The method of  claim 1 , further comprising performing an immunoassay with one or more additional antibodies that bind to ovarian and/or pancreatic cancer biomarkers. 
     
     
         9 . The method of  claim 1 , wherein the test subject is: (i) apparently healthy (ii) has a family history of ovarian or pancreatic cancer; (iii) experiencing one or more symptoms associated with ovarian cancer; or (iv) known to be suffering from ovarian or pancreatic cancer and has had or is currently undergoing treatment for ovarian or pancreatic cancer. 
     
     
         10 . The method of  claim 1 , wherein the method further comprises comparing the results of the assay from biological samples obtained from the test subject at one or more time points to assess the efficacy of a treatment regimen. 
     
     
         11 . The method of  claim 1 , wherein the anti-MLP antibody is immobilized on a substrate. 
     
     
         12 . The method of  claim 1 , wherein the immunoassay is an ELISA assay. 
     
     
         13 . A method of detecting or diagnosing the presence of a mucin-secreting cancer in a test subject comprising (a) administering to a living test subject a humanized or fully human anti-MLP antibody or antigen-binding fragment thereof that binds to an epitope in the C-terminal region of MLP, set forth as SEQ ID NO:4; and (b) detecting the presence or absence or the amount of the antibody or fragment thereof bound to MLP, wherein detection of the presence or amount of MLP in the subject indicates the presence of a mucin-secreting cancer. 
     
     
         14 . The method of  claim 13 , wherein the mucin-secreting cancer is selected from the group consisting of ovarian, pancreatic, colorectal, breast, appendiceal, lung, renal, cervical, biliary, esophageal and epithelial skin cancer. 
     
     
         15 . The method of  claim 13 , wherein the anti-MLP antibody is labeled with a detectable moiety suitable for in vivo use and step (b) comprises detecting the presence or amount of the detectable moiety. 
     
     
         16 . The method of  claim 13 , wherein method is used in an imaging, intraoperative, endoscopic or intravascular procedure. 
     
     
         17 . The method of  claim 13 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody that binds to the same epitope or competes for binding to MLP with a reference antibody selected from the group consisting of:
 the monoclonal anti-MLP antibody Clone 11 produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121699;   the monoclonal anti-MLP antibody Clone B produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121700; and   the monoclonal anti-MLP antibody clone C produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121701.   
     
     
         18 . The method of  claim 13 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody having a heavy chain variable region CDR-H3 sequence comprising an amino acid sequence set forth as SEQ ID NO:15, SEQ ID NO:35, SEQ ID NO:36 or SEQ ID NO:19, and conservative sequence modifications thereof, and having a light chain variable region CDR-L3 sequence comprising an amino acid sequence set forth as SEQ ID NO:23 or SEQ ID NO:27, and conservative sequence modifications thereof. 
     
     
         19 . The method of  claim 14 , wherein the test subject is: (i) apparently healthy (ii) has a family history of ovarian or pancreatic cancer; (iii) experiencing one or more symptoms associated with ovarian cancer; or (iv) known to be suffering from ovarian or pancreatic cancer and has had or is currently undergoing treatment for ovarian or pancreatic cancer. 
     
     
         20 . A method of treating a subject suffering from ovarian or pancreatic cancer comprising administering to an individual suffering from ovarian or pancreatic cancer a humanized or fully human anti-MLP antibody or antigen-binding fragment thereof that binds to an epitope in the C-terminal region of MLP, set forth as SEQ ID NO:4, wherein the antibody or fragment thereof is coupled to a therapeutic agent. 
     
     
         21 . The method of  claim 20 , wherein the therapeutic agent is a chemotherapeutic agent. 
     
     
         22 . The method of  claim 20 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody that binds to the same epitope or competes for binding to MLP with a reference antibody selected from the group consisting of:
 the monoclonal anti-MLP antibody Clone 11 produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121699;   the monoclonal anti-MLP antibody Clone B produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121700; and   the monoclonal anti-MLP antibody clone C produced by the hybridoma cell line deposited at the ATCC on Oct. 30, 2014 under the ATCC Designation Number PTA-121701.   
     
     
         23 . The method of  claim 20 , wherein the anti-MLP antibody or fragment thereof is a monoclonal antibody having a heavy chain variable region CDR-H3 sequence comprising an amino acid sequence set forth as SEQ ID NO:15, SEQ ID NO:35, SEQ ID NO:36 or SEQ ID NO:19, and conservative sequence modifications thereof, and having a light chain variable region CDR-L3 sequence comprising an amino acid sequence set forth as SEQ ID NO:23 or SEQ ID NO:27, and conservative sequence modifications thereof.