抗人大肠癌重组单链抗体的研制  

MAKING SINGLE-CHAIN ANTIBODY AGAINST HUMAN COLORECTAL CARCINOMA CELLS

作　　者：陈立[1] 宋今丹[1] 

机构地区：[1]中国医科大学细胞生物学卫生部重点实验室,沈阳110001

出　　处：《细胞生物学杂志》1999年第3期148-150,共3页Chinese Journal of Cell Biology

基　　金：国家"八五"科技攻关项目;85-722-18-02

摘　　要：应用重组噬菌体抗体系统制备了重组单链抗体。首先从抗人结肠癌ND-1单抗杂交瘤细胞中提取mRNA,利用反转录多聚酶链反应(RT-PCR)扩增出单抗重链可变区(V_H)及轻链可变区(V_L)片段,再通过连接DNA合成单链抗体可变区片段(ScFv),然后经双酶切后克隆到pCANTAB5E载体中,在E.coliTGI细胞中表达出噬菌体融合蛋白,用抗原阳性噬菌体感染E.coliHB2151细胞,产生单链抗体,该单链抗体既保持了原单抗的特异性,应用上又优于原单抗。A single-chain antibody against human colorectal carcinoma was synthesized using the Recombinant Phage Anti-body System. First,we used RT-PCR to amplify the VH and VL genes from the mouse hybridoma mRNA. The VH and VL genes were then cloned into the phagemid vector pCANTABSE and expressed as a fusion with a phage protein. Antigen-positive phage was used to infecte E, coli HB2151 cells. A colony was then picked and grown up to provide a seed culture for production of soluble single-chain antibody. The recombinant antibody retains its antigen-binding specificity.

关 键 词：大肠癌 单链抗体 重组 研制 

分 类 号：R735.340.3[医药卫生—肿瘤]