非复制型腺病毒介导抗-HBC单链抗体的细胞内表达  

Expression of anti-HBc single-chain variable fragment mediated by recombinant replication defective adenovirus in vitro

作　　者：汤正好[1] 余永胜[1] 马会慧[2] 江红[1] 奚敏[1] 臧国庆[1] 姚集鲁[2] 

机构地区：[1]上海交通大学附属第六人民医院感染病科,200233 [2]广州中山大学附属第三医院传染病科

出　　处：《中华肝脏病杂志》2006年第8期587-589,共3页Chinese Journal of Hepatology

摘　　要：目的 探讨非复制型腺病毒介导抗-HBc单链抗体（ScFv）细胞内表达效率，并确定是否具有特异性抗原结合活性。方法 将经细菌内同源重组并在293细胞中包装产生的携带抗-HBcScFv基因的非复制型重组腺病毒，按感染复数为10体外转染HepG2细胞，荧光显微镜下观察绿色荧光蛋白的表达；取培养上清液和细胞裂解上清液进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳和Western blot法检测HBc ScFv表达。结果 重组腺病毒Ad-ScFv感染HepG2细胞后，荧光显微镜下观察到HepG2细胞中绿色荧光蛋白；十二烷基硫酸钠聚丙烯酰胺凝胶电泳显示培养上清液和细胞裂解物中可见2．7×10^4左右的蛋白条带；Western blot显示有乙型肝炎核心抗原特异性结合活性阳性反应条带出现。结论 携带抗-HBc ScFv基因的非复制型重组腺病毒能在真核细胞中有效表达抗-HBc ScFv，并具有特异性抗原结合活性。Objectives To define the expression of single-chain variable fragment （ScFv） against hepatitis B virus core protein （HBc） mediated by recombinant replication defective adenovirus carrying the anti-HBc ScFv gene in vitro and to define the activity of anti-HBc ScFv combining HBcAg. Methods The recombinant adenoviruses carrying anti-HBc ScFv gene generated by homologous recombination in bacteria and packaged in 293 cells were transfected into HepG2 cells, and the anti- HBc ScFv was detected using SDS-PAGE and Western blot. Results Green fluorescent protein （GFP） was observed in HepG2 cells after the transfection. SDS-PAGE displayed a protein strap about 2.7×10^4, and the result of Western blot displayed a positive reactive strap. Conclusion Anti-HBc ScFv can be expressed in cells mediated by recombinant replication defective adenovirus carrying the anti-HBc ScFv gene.

关 键 词：腺病毒 肝炎病毒 乙型 核心蛋白 单链抗体 表达 

分 类 号：R392[医药卫生—免疫学]