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 
         X is selected from the group consisting of
 (CH 2 ) n  with n being an integer from 1 to 20, 
 [(CH 2 ) p —O] k —(CH 2 ) m  with p being 2 or 3, with m being 2 or 3, and with k being an integer from 1 to 30, and 
 [(CH 2 ) r —CONH] s —(CH 2 ) t  with r being an integer from 1 to 5, with t being an integer from 0 to 5, and with s being an integer from 1 to 5 
 and 
 
         R is selected from the group consisting of H, OH, COOH, NH 2 , an azide group, a maleimide group, and Z, 
         wherein 
         Z is A or B, 
         wherein A is M-L with M being selected from (i) a hapten which does not contain a biotin moiety and (ii) a polypeptide, and L being a linker, 
         wherein B is 
       
       
         
           
           
               
               
           
         
         with M being selected from (i) a hapten which does not contain a biotin moiety and (ii) a polypeptide, and L being a linker, whereby the nitrogen atom marked with an asterisk is covalently bound to an adjacent CH 2  group of X. 
       
     
     
         2 . The antibody of  claim 1  further characterized in that it does not bind to a compound of Formula II, also depicted in  FIG. 3A . 
       
         
           
           
               
               
           
         
       
     
     
         3 . The monoclonal antibody according to  claim 1 , wherein the binding affinity of the monoclonal antibody to any one compound of the group of compounds selected from 
       
         
           
           
               
               
           
         
       
       is higher by a factor of at least 50 than the binding affinity to the compound of Formula II. 
     
     
         4 . The monoclonal antibody according to  claim 3 , wherein the binding affinity of the monoclonal antibody to any one compound of the group of compounds selected from Formula III A, Formula III B, and Formula III C, 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 (strept)avidin coated solid phase,
 followed by measuring the analyte bound to coated (strept)avidin on the solidphase 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 . 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 or a labeled (strept)avidin.   
     
     
         8 - 13 . (canceled)