Anti-apo-B-48 monoclonal antibody, hybridoma producing the same, and method for measuring apo-B-48 using the same

Abstract

A novel monoclonal antibody which specifically binds to apo-B-48 is disclosed. The monoclonal antibody specifically binds to apo-B-48 but does not bind to apo-B-100.

We claim:  
     
         1 . A monoclonal antibody which specifically binds to apo-B-48 and which does not specifically bind to apo-B-100.  
     
     
         2 . The monoclonal antibody according to  claim 1 , of which corresponding epitope resides in C-terminal region of apo-B-48.  
     
     
         3 . The monoclonal antibody according to  claim 2 , wherein said C-terminal region is 1st to 20th amino acids from the C-terminal of apo-B-48.  
     
     
         4 . The monoclonal antibody according to  claim 3 , wherein said corresponding epitope contains the C-terminal of apo-B-48.  
     
     
         5 . The monoclonal antibody according to  claim 1 , which is produced by hybridoma B48-151 deposited with National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology under accession No. FERM BP-6473 or variants thereof.  
     
     
         6 . The monoclonal antibody according to  claim 5 , which is produced by said hybridoma B48-151.  
     
     
         7 . A hybridoma which produces said monoclonal antibody according to  claim 1 .  
     
     
         8 . The hybridoma according to  claim 7 , which is produced by immunizing a mammal with a peptide derived from apo-B-48, recovering antibody-producing cells from said mammal, fusing the antibody-producing cells with myeloma cells, and selecting a hybridoma which produces said monoclonal antibody according to  claim 1 .  
     
     
         9 . The hybridoma according to  claim 8 , wherein said peptide comprises C-terminal region of apo-B-48.  
     
     
         10 . The hybridoma according to  claim 9 , wherein said peptide consists essentially of not less than  5  to not more than  20  amino acids from the C-terminal of apo-B-48.  
     
     
         11 . The hybridoma according to  claim 10 , wherein said peptide is a conjugate of a peptide consisting essentially of 1st-6th amino acids from the C-terminal of apo-B-48 and a carrier protein.  
     
     
         12 . The hybridoma according to  claim 8 , wherein said peptide is conjugated with said carrier protein through cystein residue attached to the N-terminal of said peptide.  
     
     
         13 . The hybridoma according to  claim 7 , which is hybridoma B48-151 deposited with National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology under accession No. of FERM BP-6473 or variants thereof.  
     
     
         14 . The hybridoma according to  claim 13 , which is hybridoma B48-151.  
     
     
         15 . A method for measuring apo-B-48 in a test sample, comprising utilizing antigen-antibody reaction between apo-B-48 in said test sample and said monoclonal antibody according to any one of claims  1 - 6 .  
     
     
         16 . The method according to  claim 15 , wherein said monoclonal antibody is immobilized on a solid carrier, and apo-B-48 immobilized on said carrier through said monoclonal antibody is measured after said antigen-antibody reaction.  
     
     
         17 . The method according to  claim 16 , which is carried out by ELISA.  
     
     
         18 . The method according to  claim 15 , wherein the apo-B-48 in said test sample is subjected to a treatment by which the corresponding epitope of said monoclonal antibody is exposed before or simultaneously with said antigen-antibody reaction.  
     
     
         19 . The method according to  claim 18 , wherein said treatment comprises treating said sample with a surfactant.  
     
     
         20 . The method according to  claim 19 , wherein said surfactant is a nonionic surfactant.  
     
     
         21 . A reagent for immunoassay for measuring apo-B-48 comprising said monoclonal antibody according to any one of claims  1 - 6 .  
     
     
         22 . The reagent according to  claim 21 , wherein said monoclonal antibody is immobilized on a solid carrier.  
     
     
         23 . A kit for immunoassay for measuring apo-B-48 comprising said monoclonal antibody according to any one of claims  1 - 6 .  
     
     
         24 . The kit according to  claim 23 , wherein said monoclonal antibody is immobilized on a solid carrier.