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

本研究的最终目的是利用新合成的含有BMZ抗原蛋白表位的多肽抗原检测大疱性类天疱疮（BP）患者的抗基底膜带（BMZ）抗体。K.和联合S. BP患者我们发现，来自不同大陆的BMZ抗体在抗原表位方面具有相似的异质性，如通过免疫荧光、免疫印迹和免疫沉淀所评估的。然后，我们利用重组BP抗原蛋白对BP血清进行了表征，并测试了应用ELISA技术的可能性。同时，我们利用编码人抗BMZ单克隆抗体5E羧基端约一半的cDNA克隆的缺失突变体，确定了人抗BMZ单克隆抗体5E的230 kD BP抗原表位。230 kD BP抗原蛋白。5E抗原表位定位于Bgl 11-StuI片段编码的114个氨基酸残基。从这些研究中得到以下结论：（1）BEIZ抗原中存在几个表位，（2）对于ELISA技术仅检测230 kD抗原，需要3个以上的合成肽来覆盖整个表位。需要更多的调查才能到达最终目的地。

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.