肝胆胰外科杂志2011，Vol.23Issue(4)：325-328,335,5.
抗人AFP单链抗体的制备、鉴定及其免疫学活性的测定
Construction, identification and determination of immunologic activity of murine anti-AFP ScFv
摘要
Abstract
Objective To construct and identify the single-chain variahle fragment (scFv) of monoclonal antihody against murine AFP and analyze the immunologic activity of recombinant ScFv protein. Methods The VL and VH genes were cloned by RT-PCR from anti-murine AFP mAb excreted by hybridoma cells. VH-linker-VL fragment (ScFv) was constructed and cloned into vector pUC19. The purified pUC19-ScFv plasmid was transformed into BL21 cells and induced hy IPTG, the expression and activity of ScFv protein were analyzed with SDS-PAGE and ELISA. Results BL21 cells could express recombinant ScFv induced by IPTG.SDS-PAGE analysis revealed that the ScFv was expressed in inclusion body. Indirect ELISA results indicated that the expressed ScFv could bind specifically to recombinant protein of murine AFP. and the epitope recognized by the ScFv was same as the parental mAb of AFP. Conclusion Expression vector encoding murine anti-AFP ScFv is successfully constructed and it can efficaciously express ScFv with immune activity and specificity.关键词
甲胎蛋白/单链抗体/免疫学活性Key words
alpha fetoprotein/ single-chain variable fragment/ immunologic activity分类
医药卫生引用本文复制引用
牛坚,张业伟,刘斌..抗人AFP单链抗体的制备、鉴定及其免疫学活性的测定[J].肝胆胰外科杂志,2011,23(4):325-328,335,5.基金项目
江苏省高校自然科学研究资助项目(09kjb320018). （09kjb320018）
