抗HBsAg/RBC双特异噬菌体抗体的构建  

Construction of bi-specific phage antibody against HBsAg and RBC

作　　者：刘晓琳[1] 王琰[1] 张达矜[1] 陈宇萍[1] 

机构地区：[1]海军总医院,北京100037

出　　处：《中华微生物学和免疫学杂志》2003年第11期907-909,共3页Chinese Journal of Microbiology and Immunology

基　　金：全军医药卫生"九五"重点项目 (96Z0 0 4)

摘　　要：目的　通过不同外壳蛋白的展示 ,构建抗HBsAg RBC双特异噬菌体抗体。方法　通过DNA重组技术 ,将抗HBsAgFab与噬菌体基因 8融合 ,抗RBCScFv与噬菌体基因 3融合 ,这 2种融合基因以不同的启动子驱动 ,并克隆于同一噬菌体表达载体上 ,得到双特异噬菌体抗体表达载体 ,转化大肠杆菌后获取噬菌体抗体上清 ,用ELISA和红细胞凝集试验检测其双特异活性。结果　ELISA和RBC凝集实验证明 ,本方法可形成双特异噬菌体抗体 ,可使含有HBsAg的RBC悬液产生凝集。用展示性能得到提高的蛋白 8变种取代野生型蛋白 8可提高双特异噬菌体抗体的活性。结论　HBsAb和RBC两种抗体分子能够通过不同噬菌体外壳蛋白同时展示于同一噬菌体表面 ,形成双特异噬菌体抗体。可用于人外周血HBsAg的快速血凝法检测。Objective To construct an anti-HBsAg/RBC bi-specific phage antibody using phage display technique through different coat protein mediated display. Methods Anti-HBsAg Fab and anti-RBC ScFv genes were fused to phage gene 8 and gene 3 respectively. The two fused genes were driven by different promoters and were cloned into a single expression vector. The resulting vector was transformed into E.coli and phage antibodies were rescued by helper phage superinfection. ELISA and RBC agglutination test were used to check the bi-specific activity of the phage antibody. Results The resulting phage antibodies could bind to both HBsAg and RBCs. When HBsAg was present the phages could cause agglutination of RBCs, thus proving the bi-specific nature of the phage antibody. The bi-specific activity could be improved by replacing the wild type gene 8 with gene 8 mutants that mediated more efficient display of antibody fragments. Conclusion The anti-HBsAg Fab and anti-RBC ScFv can be displayed on a single phage particle by different coat protein mediated display. The resulting bi-specific phage antibody can be used to detect HBsAg in form of a quick RBC agglutination test.

关 键 词：双特异噬菌体抗体 噬菌体展示 红细胞凝集试验 乙肝表面抗原 

分 类 号：R392[医药卫生—免疫学]