Detection of antigen epitope for bullous pemphigois antigenic protein and the development of autoantibody detection using synthetic polypeptide

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The final destination of this developmental scientific research was to detect anti-basement membrane zone (BMZ) antibodies from patients with bullous pemphigoid (BP) using a newly synthesized polypeptide antigen that contains epitope for BMZ antigen protein.To approach this purpose, we have collected as many sera as possible from Japan, U. K. and U. S. patients with BP. We have found that the BMZ antibodies from different continents are similarly heterogenous in terms of antigenic epitope as assessed by immunofluorescence, immunoblotting and immunoprecipitation. Then, we have characterized BP sera by use of recombinant BP antigen proteins and tested the possibility of applying ELISA technique. At the same time we specified the epitope of the"230 kD BP antigen for human anti-BMZ monoclonal antibody 5E by means of deletion mutants of the mouse CDNA Clone that encodes for about carboxyl-terminal half of t. he 230 kD BP antigen protein. The epitope for 5E was found to localize in the 114 amino acid residues encoded by Bgl 11 - StuI fragment of the CDNA. From these studies the following conclusions were obtained ; (1) there are several epitopes present in the BEIZ antigens and (2) more than 3 synthetic peptides are necessary to cover the entire epitopes for ELISA technique to detect 230 kD antigen only. More investigations are needed to reach the final destination.