Assays for the detection of anti-tnf drugs and autoantibodies

Abstract

The present invention provides assays for detecting and measuring the presence or level of anti-TNFα drug therapeutics and autoantibodies in a sample. The present invention is useful for optimizing therapy and monitoring patients receiving anti-TNFα drug therapeutics to detect the presence or level of autoantibodies (e.g., HACA and/or HAHA) against the drug.

1 - 84 . (canceled) 
     
     
         85 . A method for determining the presence or level of a therapeutic antibody in a sample, the method comprising:
 (a) contacting a labeled antigen of the therapeutic antibody with a sample having a therapeutic antibody to form a labeled complex with the therapeutic antibody;   (b) subjecting the labeled complex to size exclusion chromatography to separate the labeled complex from free labeled antigen and to detect an amount of the labeled complex and an amount of the free labeled antigen; and   (c) comparing the amount of the labeled complex and the amount of the free labeled antigen detected in step (b) to a standard curve of known amounts of the therapeutic antibody,   
       thereby determining the presence or level of the therapeutic antibody. 
     
     
         86 . The method of  claim 85 , wherein the size exclusion chromatography comprises size exclusion-high performance liquid chromatography (SE-HPLC). 
     
     
         87 . The method of  claim 85 , wherein step (a) comprises a liquid phase reaction. 
     
     
         88 . The method of  claim 85 , performed without a wash step. 
     
     
         89 . The method of  claim 85 , wherein the standard curve is generated by incubating the labeled antigen with known amounts of the therapeutic antibody. 
     
     
         90 . The method of  claim 85 , wherein the labeled complex is eluted first, followed by the free labeled antigen. 
     
     
         91 . The method of  claim 85 , wherein the sample is obtained from a subject receiving therapy with the therapeutic antibody. 
     
     
         92 . The method of  claim 85 , wherein the sample is a serum sample. 
     
     
         93 . The method of  claim 85 , wherein the labeled antigen comprises a fluorescent label. 
     
     
         94 . The method of  claim 93 , wherein the fluorescent label comprises an Alexa Fluor® dye. 
     
     
         95 . The method of  claim 85 , wherein the therapeutic antibody is monoclonal. 
     
     
         96 . The method of  claim 85 , wherein the therapeutic antibody is humanized. 
     
     
         97 . The method of  claim 85 , wherein the therapeutic antibody treats an inflammatory bowel disease. 
     
     
         98 . The method of  claim 97 , wherein the inflammatory bowel disease comprises Crohn's disease or ulcerative colitis. 
     
     
         99 . The method of  claim 85 , wherein the therapeutic antibody is an IgG1 antibody. 
     
     
         100 . The method of  claim 85 , wherein the therapeutic antibody recognizes an anti-cell adhesion molecule. 
     
     
         101 . The method of  claim 85 , wherein the therapeutic antibody recognizes an α4-integrin. 
     
     
         102 . The method of  claim 85 , wherein the therapeutic antibody is an anti-α4β7-integrin antibody. 
     
     
         103 . The method of  claim 85 , wherein the therapeutic antibody is a humanized IgG1 anti-α4β7-integrin monoclonal antibody. 
     
     
         104 . The method of  claim 85 , wherein the therapeutic antibody is MLN-02.