抗志贺毒素ⅡB亚单位单克隆抗体识别表位的初步鉴定  

Elementary characterization on epitopes of anti-shiga toxinⅡB subunit monoclonal antibody

作　　者：陈洪章[1] 曾浩[1] 毛旭虎[1] 罗萍[1] 余抒[1] 邹全明[1] 

机构地区：[1]第三军医大学医学检验系临床微生物学及免疫学教研室,重庆市生物制药工程技术研究中心,重庆400038

出　　处：《第三军医大学学报》2007年第19期1831-1833,共3页Journal of Third Military Medical University

基　　金：国家高技术研究发展"863"计划(2006AA02Z405)~~

摘　　要：目的初步确定抗志贺毒素ⅡB亚单位(Stx2B)单克隆抗体(McAb)1H9、1H10、3E5识别的抗原表位。方法运用DNAstar Protean软件,对Stx2B(1～72aa)全抗原进行表位预测;采用截短法分段构建重组质粒Stx2B50(23～72aa),Stx2B54(1～54aa)。用IPTG诱导表达融合蛋白。对表达产物进行SDS-PAGE蛋白电泳及Westernblot分析。结果1H9、1H10能够识别Stx2B和Stx2B50;3E5能够识别Stx2B、Stx2B50和Stx2B54。结论McAb1H9、1H10识别的抗原表位位于55～72aa,McAb3E5识别的抗原表位位于23～54aa。Objective To identify the epitope domain of monoclonal antibody 1H9,1H10,and 3E5 against shiga toxin Ⅱ B subunit（Stx2B）. Methods The epitopes of Stx2B （1 -72 aa） were predicted by the DNAstar Protean software. The recombinant plasmids of Stx2B50 （23 -72 aa）, Stx2B54 （1 -54 aa） were constructed and then transformed to E. coli BL21 （ DE3 ） for expression under IPTG induction. The proteins were identified by SDS-PAGE and Western blotting. Results The monoclonal antibody 1H9 and 1Hi0 recognized antigen of Stx2B and Stx2B 50, while 3E5 recognized Stx2B, Stx2B50, and Stx2B54. Conclusion The epitope of mono- clonal antibody 1H9 and 1H10 is located at 55 -72 aa and the epitope of 3E5 is located at 23 -54 aa.

关 键 词：志贺毒素ⅡB亚基 抗原表位 鉴定 

分 类 号：R392.12[医药卫生—免疫学] R394.33[医药卫生—基础医学]