Novel biotin-specific monoclonal antibody and use thereof

Abstract

The present invention relates to a monoclonal antibody capable of binding to biotin. In one embodiment the monoclonal antibody according to the invention also does not bind to a biotin moiety on a biotinylated molecule, wherein the biotin moiety is attached to the molecule via the carbon atom of the carboxyl function of the valeric acid moiety of biotin. Also disclosed is a method for generation of an antibody as disclosed herein. The monoclonal antibody according to the invention is of specific use in a method for measuring an analyte in a sample, wherein a (strept)avidin/biotin pair is used to bind a biotinylated analyte specific binding agent to a (strept)avidin coated solid phase.

1 . A monoclonal antibody specifically binding the compound of Formula I, 
       
         
           
           
               
               
           
         
         characterized in that it also binds to biotin, 
         wherein 
         Y is CH 2  or C═O, 
         X is O or (CH 2 ) n  and n is an integer from 1 to 20, and 
         R is either H, or Z-L, 
         wherein 
         L is a linker, and 
         Z is selected from a hapten not containing a biotin moiety and a polypeptide. 
       
     
     
         2 . The antibody of  claim 1  further characterized in that it does not bind to a compound of Formula II 
       
         
           
           
               
               
           
         
       
     
     
         3 . The monoclonal antibody of  claim 1 , wherein the binding affinity of the monoclonal antibody to the compound of Formula III is higher by a factor of at least 50 than the binding affinity to the compound of Formula II, 
       
         
           
           
               
               
           
         
       
     
     
         4 . The monoclonal antibody of  claim 3 , wherein the binding affinity of the monoclonal antibody to the compound of Formula III is higher by a factor of at least 500 than the binding affinity to the compound of Formula II. 
     
     
         5 . A method for measuring an analyte in a sample, wherein a (strept)avidin/biotin binding pair is used to bind a biotinylated analyte specific binding agent to a (strept)avidin coated solid phase,
 the method comprising adding to the sample   a) an antibody according to  claim 1 ,   b) a biotinylated analyte specific binding agent,   c) a streptavidin coated solid phase,   followed by   measuring the analyte bound to the solid phase via (strept)avidin and biotinylated analyte specific binding agent.   
     
     
         6 . The method of  claim 5 , wherein step 5 (a) and optionally also step 5 (b) is performed before step 5 (c). 
     
     
         7 . (canceled) 
     
     
         8 . An immunoassay test kit comprising at least
 a) an antibody according to  claim 1 ,   b) a biotinylated analyte specific binding agent, and   c) a (strept)avidin coated solid phase.   
     
     
         9 - 14 . (canceled)