抗人大肠癌重组单链抗体的研制

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。

MAKING SINGLE-CHAIN ANTIBODY AGAINST HUMAN COLORECTAL CARCINOMA CELLS

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.