抗乙型肝炎表面抗原单链抗体(ScFv)的原核表达、纯化及鉴定  

Expression,purification and identification of the anti-HBsAg ScFv

作　　者：雷清[1] 刘鹏[1] 郭敏[1] 蒋琳[1] 

机构地区：[1]兰州生物制品研究所,兰州730046

出　　处：《微生物学免疫学进展》2008年第4期36-39,共4页Progress In Microbiology and Immunology

基　　金：甘肃省自然基金资助项目(0710RJZA079)

摘　　要：从抗HBsAg鼠单抗的杂交瘤细胞提取RNA,经反转录得到cDNA,进一步扩增得到鼠重链可变区基因(VH)和轻链可变区基因(VL),按VH-linker-VL的结构将VH、VL基因拼接成单链抗体(scFv)基因;经测序正确后进一步构建了表达重组体p26HBSc并在E.coli中表达,得到一约30kD的外源蛋白;纯化后经ELISA检测与HBsAg有较高的亲和力活性,为下一步的人源化改造奠定了基础。Obtain RNA from mouse anti-HBsAg hybridoma cell line, cDNA was got by RT, then the VH and VL gene of mouse anti-HBsAg was obtained for further amplification. According to the structure of VH-linker-VL, put VH and VL fragments together to form single chain Fv. Construct p26HBSc after sequencing and expressed in E. coli. Intrested protein was expressed about 30kD, It showed that it had higher affinity with HBsAg, so it is of potentially important for its humanization in the future.

关 键 词：单链抗体 连接肽 基因表达 

分 类 号：R392[医药卫生—免疫学]