[抗木瓜凝乳蛋白酶单链抗体噬菌体展示文库的构建与鉴定]

[Construction and identification of anti-chymopapain scFv phage display library].

作者信息

Wang Jian-ping, Zhao Shu-jin, Wang Jie, Wu Jie, Yang Tai-cheng, Yang Jing

机构信息

Department of Bioengineering, South China University of Technology, Guangzhou 510640, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Nov;20(6):689-92.

PMID:15555436

Abstract

AIM

To construct phage display library of anti-chymopapain scFv.

METHODS

V(H) and V(L) gene repertoires were amplified from splenocyte mRNA by RT-PCR and joined by a (Gly(4)ser)3 linker to obtain scFv genes. The scFv genes were then cloned into phagemid pFAB5C to construct phage display library. Affinity selection and ELISA were used to identify specific phage antibody to chymopapain.

RESULTS

After 4 rounds of panning, high affinity scFv was obtained.

CONCLUSION

Phage display library of anti-chymopapain scFv was successfully constructed, and scFv with binding ability to chymopapain was obtained.

摘要

目的

构建抗木瓜凝乳蛋白酶单链抗体片段（scFv）噬菌体展示文库。

方法

通过逆转录聚合酶链反应（RT-PCR）从脾细胞信使核糖核酸（mRNA）中扩增重链可变区（V(H)）和轻链可变区（V(L)）基因库，并用(Gly(4)ser)3接头连接以获得scFv基因。然后将scFv基因克隆到噬菌粒pFAB5C中构建噬菌体展示文库。采用亲和筛选和酶联免疫吸附测定（ELISA）鉴定针对木瓜凝乳蛋白酶的特异性噬菌体抗体。