He4 monoclonal antibodies and methods for their use

Abstract

Compositions and methods for diagnosing ovarian cancer in a patient and for identifying patients with an increased likelihood of having ovarian cancer are provided. The compositions include novel monoclonal antibodies, and variants and fragments thereof, that specifically bind to HE4. Monoclonal antibodies having the binding characteristics of an HE4 antibody of the invention are further provided. Hybridoma cell lines that produce an HE4 monoclonal antibody of the invention are also disclosed herein. The compositions find use in diagnostic methods as well as in screening methods for identifying patients having an increased likelihood of having ovarian cancer. Kits comprising one or more of the disclosed HE4 monoclonal antibodies and for practicing the methods of the invention are further provided. Polypeptides comprising the amino acid sequence for an HE4 epitope and methods of using these polypeptides in the production of antibodies are also encompassed by the present invention.

1 . A monoclonal antibody that is capable of specifically binding to HE4, wherein the antibody is selected from the group consisting of:
 (a) the monoclonal antibody produced by the hybridoma cell line 363A71.1, deposited with the ATCC as Patent Deposit No. PTA-8195;   (b) a monoclonal antibody that binds to an epitope comprising the amino acid sequence set forth in SEQ ID NO:12 or 13; and   (c) a monoclonal antibody that is an antigen binding fragment of a monoclonal antibody of (a) or (b), wherein the fragment retains the capability of specifically binding to HE4.   
     
     
         2 . A hybridoma cell line capable of producing a monoclonal antibody of  claim 1 . 
     
     
         3 . A kit for diagnosing ovarian cancer in a patient comprising:
 a) a capture antibody immobilized on a solid support, wherein the capture antibody is a first HE4 antibody; and   b) a tag antibody, wherein the tag antibody is a second HE4 antibody that is labeled with a detectable substance;   wherein said first or said second HE4 antibody is the monoclonal antibody of  claim 1 .   
     
     
         4 . A kit for diagnosing ovarian cancer or for identifying patients with an increased likelihood of having ovarian cancer comprising at least one monoclonal antibody according to  claim 1 . 
     
     
         5 . A method for diagnosing ovarian cancer in a patient comprising:
 a) incubating a body sample from the patient with a capture antibody immobilized on a solid support, wherein the capture antibody is a first HE4 antibody;   b) detecting bound HE4 by contacting the solid support with a tag antibody, wherein the tag antibody is a second HE4 antibody that is labeled with a detectable substance; and   c) quantitating the bound tag antibody, and thereby determining the level of bound HE4 protein;   wherein said first or said second HE4 antibody is selected from the group consisting of:
 i) the monoclonal antibody of  claim 1 ; 
 i) the monoclonal antibody produced by the hybridoma cell line 363A90.1, deposited with the ATCC as Patent Deposit No. PTA-8196; 
 (iii) a monoclonal antibody that binds to the amino acid sequence set forth in SEQ ID NO:11; and, 
 (iv) a monoclonal antibody that is an antigen binding fragment of a monoclonal antibody of (ii) or (iii), wherein the fragment retains the capability of specifically binding to HE4. 
   
     
     
         6 . The method of  claim 5  further comprising comparing the level of bound HE4 protein in the patient body sample to a threshold level, wherein the threshold level is determined by measuring the level of bound HE4 protein in samples from patients that do not have ovarian cancer. 
     
     
         7 . The method of  claim 5 , wherein the capture antibody is the monoclonal antibody produced by the hybridoma cell line 363A90.1, deposited with the ATCC as Patent Deposit No. PTA-8196, and wherein the tag antibody is the monoclonal antibody produced by the hybridoma cell line 363A71.1, deposited with the ATCC as Patent Deposit No. PTA-8195. 
     
     
         8 . A screening method for identifying patients with an increased likelihood of having ovarian cancer, the method comprising:
 a) performing a first assay step comprising detecting the expression of HE4 in a body sample and determining if HE4 is overexpressed; and   b) performing a second assay step if HE4 is determined to be overexpressed in step (a), the second assay step comprising detecting the expression of a second biomarker or a second panel of biomarkers in the body sample and determining if the second biomarker or second panel of biomarkers is overexpressed, wherein overexpression of both HE4 and the second biomarker or panel of biomarkers is indicative of an increased likelihood of the patient having ovarian cancer;   wherein HE4 expression is detected by contacting the body sample with a monoclonal antibody capable of specifically binding to HE4, wherein the antibody is selected from the group consisting of:
 i) the monoclonal antibody of  claim 1 ; 
 ii) the monoclonal antibody produced by the hybridoma cell line 363A90.1, deposited with the ATCC as Patent Deposit No. PTA-8196; 
 (iii) a monoclonal antibody that binds to the amino acid sequence set forth in SEQ ID NO:11; and, 
 (iv) a monoclonal antibody that is an antigen binding fragment of a monoclonal antibody of (ii) or (iii), wherein the fragment retains the capability of specifically binding to HE4. 
   
     
     
         9 . The method of  claim 8 , wherein HE4 expression is detected by contacting the body sample with at least two monoclonal antibodies that specifically bind to HE4, wherein a first antibody is the monoclonal antibody produced by the hybridoma cell line 363A90.1, deposited with the ATCC as Patent Deposit No. PTA-8196, and a second antibody is the monoclonal antibody produced by the hybridoma cell line 363A71.1, deposited with the ATCC as Patent Deposit No. PTA-8195. 
     
     
         10 . The method of  claim 8 , wherein the second panel of biomarkers is selected from the group consisting of a panel comprising CA125, glycodelin, Muc-1, PAI-1, and PLAU-R, a panel comprising CA125 and PAI-1, a panel comprising CA125, glycodelin, PAI-1, and MMP-7, a panel comprising CA125, glycodelin, PAI-1, and PLAU-R, a panel comprising CA125, glycodelin, PAI-1, and PLAU-R, a panel comprising glycodelin, Muc-1, PLAU-R, and inhibin A, a panel comprising CA125, Muc-1, glycodelin, PAI-1, and PLAU-R, and a panel comprising CA125, MMP-7, glycodelin, and PLAU-R. 
     
     
         11 . An isolated polypeptide consisting of an epitope for binding an HE4 monoclonal antibody, wherein the polypeptide consists of an amino acid sequence selected from the group consisting of:
 (a) the amino acid sequence set forth in SEQ ID NO:11, 12, or 13; and,   (b) an amino acid sequence having at least 90% sequence identity to SEQ ID NO:11, 12, or 13, wherein the polypeptide has antigenic activity.   
     
     
         12 . An isolated nucleic acid molecule that encodes a polypeptide consisting of an epitope for binding an HE4 monoclonal antibody, the nucleic acid molecule consisting of a nucleotide sequence selected from the group consisting of:
 (a) a polynucleotide encoding the amino acid sequence set forth in SEQ ID NO:11, 12, or 13; and,   (b) a polynucleotide encoding an amino acid sequence of a polypeptide having at least 90% sequence identity to SEQ ID NO:11, 12, or 13, wherein the polynucleotide encodes a polypeptide having antigenic activity.   
     
     
         13 . A method for producing an HE4 antibody comprising immunizing an animal with a polypeptide according to  claim 12 . 
     
     
         14 . A method for producing an HE4 monoclonal antibody comprising:
 (a) immunizing an animal with a polypeptide according to  claim 12  under conditions to elicit an immune response;   (b) isolating antibody-producing cells from the animal;   (c) fusing the antibody-producing cells with immortalized cells in culture to form monoclonal antibody-producing hybridoma cells;   (d) culturing the hybridoma cells; and,   (e) isolating monoclonal antibodies from culture.