Antibodies as a cancer diagnostic

Abstract

Method and kits are provided for the detection and diagnosis of metastatic disease. More particularly, the methods and kits employ compounds that can detect EphA2, a specific epithelial cell tyrosine kinase that is overexpressed in metastatic tumor cells. In one embodiment the compound is an antibody capable of binding to an epitope of EphA2.

1 - 66 . (canceled) 
     
     
         67 . A method for detecting the presence of metastatic or potentially metastatic cells in a cell population comprising the steps of:
 contacting the cells with a compound capable of specific binding to EphA2 to allow compound binding to EphA2; and   detecting compound-EphA2 binding.   
     
     
         68 . The method of  claim 67  further comprising lysing at least a portion of the cell population, and contacting the lysed cells with the compound. 
     
     
         69 . The method of  claim 67  wherein the compound comprises an antibody specific for a domain of the EphA2 protein, a natural or artificial ligand, a peptide, an anti-sense molecule, an ATP analogue, or a small molecule capable of specifically targeting EphA2. 
     
     
         70 . The method of  claim 67  wherein the compound is a monoclonal antibody. 
     
     
         71 . The method of  claim 70  further comprising lysing at least a portion of the cell population, and contacting the lysed cells with the antibody, wherein the antibody binds to an intracellular epitope of EphA2. 
     
     
         72 . The method of  claim 70  wherein the antibody is produced by hybridoma cell line D7 (ATCC number PTA 2755). 
     
     
         73 . The method of  claim 67  further comprising detecting whether EphA2 is overexpressed, wherein overexpression of EphA2 relative to normal cells is indicative of the presence of metastatic or potentially metastatic cells in the cell population. 
     
     
         74 . The method of  claim 67  wherein the compound is labeled with a detectable label, and the detecting step includes detecting the label. 
     
     
         75 . The method of  claim 67  wherein the cell population is selected from the group consisting of breast, kidney, prostate, lung, colon and epithelial cells. 
     
     
         76 . The method of  claim 67  wherein the cell population comprises cells from a tissue biopsy. 
     
     
         77 . The method of  claim 67  wherein the cell population comprises cells from a body fluid. 
     
     
         78 . The method of  claim 77  wherein the body fluid is selected from the group consisting of blood, plasma, spinal fluid, saliva, and urine. 
     
     
         79 . The method of  claim 67  further comprising detecting whether the EphA2 is phosphorylated; wherein detection of unphosphorylated EphA2 is indicative of the presence of metastatic cancer cells in the cell population. 
     
     
         80 . The method of  claim 79  wherein detecting whether the EphA2 is phosphorylated comprises contacting the EphA2 with an anti-phosphotyrosine antibody to allow binding of the anti-phosphotyrosine antibody to EphA2; and detecting anti-phosphotyrosine antibody-EphA2 binding. 
     
     
         81 . The method of  claim 67  further comprising detecting whether EphA2 is localized to sites of cell-cell contact; wherein detecting that EphA2 is not localized to sites of cell-cell contact is indicative of the presence of metastatic cancer cells in the cell population. 
     
     
         82 . A method for distinguishing between metastatic and non-metastatic cancers comprising assaying a population of cancer cells for EphA2 expression, wherein assaying the cell population comprises:
 contacting the cells with a compound capable of binding to EphA2 to allow binding of the compound to EphA2; and   detecting compound-EphA2 binding;   
       wherein EphA2 overexpression is indicative of metastatic cancer, and absence of EphA2 expression is indicative of non-metastatic cancer. 
     
     
         83 . The method of  claim 82  wherein the compound is an antibody. 
     
     
         84 . An antibody which specifically binds to an intracellular epitope of EphA2. 
     
     
         85 . The antibody of  claim 84  bound to a detectable label. 
     
     
         86 . The antibody of  claim 84  which is a monoclonal antibody.