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 selected from the group consisting of O, S, and CH 2 , 
         X is selected from
 (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, H 2 N, HO, an azide group, a maleimide group, and Z, 
         wherein 
         Z is A or B, 
         wherein A is M-L 
         and 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  that does not bind to a compound of Formula II 
       
         
           
           
               
               
           
         
       
     
     
         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: Formula IIIA 
       
         
           
           
               
               
           
         
         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 IIIA, Formula IIIB and Formula IIIC
 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 solid phase 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 or a labeled (strept)avidin.   
     
     
         9 - 14 . (canceled)