从大容量噬菌体抗体库中筛选人源性抗角蛋白scFv

首都医科大学学报 ›› 2007, Vol. 28 ›› Issue (1): 71-74.

从大容量噬菌体抗体库中筛选人源性抗角蛋白scFv

张国民¹, 乔媛媛², 陈宇萍², 吕喆¹, 安云庆¹

1. 首都医科大学微生物学和免疫学教研室;2. 海军总医院中心实验科

收稿日期:2006-04-11 修回日期:1900-01-01 出版日期:2007-02-24 发布日期:2007-02-24
通讯作者: 安云庆

Cloning of Anti-keratin ScFv From a Human Large Phage Antibody Library

Zhang Guomin¹, Qiao Yuanyuan², Chen Yuping², Lü Zhe¹, An Yunqing¹

1. Department of Microbiology and Immunology, Capital Medical University;2. Central Lab of Navy General Hospital

Received:2006-04-11 Revised:1900-01-01 Online:2007-02-24 Published:2007-02-24

摘要/Abstract

摘要： 目的从大容量噬菌体抗体库中筛选人源性抗角蛋白单链抗体(scFv),测定其特异性结合活性并进行基因序列分析。方法以混合分子量人表皮角蛋白包被固相,对构建的大容量噬菌体抗体库进行筛选,经4轮"吸附—洗脱—扩增"后,挑取单克隆,用ELISA法测定特异性结合活性,并对阳性克隆抗体基因进行DNA指纹分析及序列同源性分析。结果经4轮筛选,获得9株可表达抗人角蛋白单链抗体的克隆,酶切鉴定正确后,经DNA指纹分析及序列同源性分析证明为不同的抗体基因。结论利用噬菌体抗体库技术获得了人源性抗角蛋白单链抗体,为临床应用研究提供了具有更广阔应用前景的人源小分子抗体。

关键词: 噬菌体抗体库, 角蛋白, 单链抗体

Abstract: Objective To clone human single chain antibodies(scFv) against keratin from a large phage antibody library and analyze the specificity of antigen binding activity and the homology of gene sequences. Methods A human large phage antibody library was panned for four times. The specificity of the antigen binding activity of the selected clones was identified by ELISA. Positive clones were analyzed by DNA fingerprint and sequencing. Results After 4 rounds of "adhesion-elution-amplification" panning, 9 clones expressing anti-keratin scFv were obtained. DNA fingerprint and sequence analysis showed that they had different antibody V region encoding genes. Conclusion Human anti-keratin scFv is obtained from a large phage antibody library, which will benefit to the application of anti-keratin antibody to clinical research.

Key words: phage antibody libraries, keratin, scFv

中图分类号:

R392.12

张国民;乔媛媛;陈宇萍;吕喆;安云庆. 从大容量噬菌体抗体库中筛选人源性抗角蛋白scFv[J]. 首都医科大学学报, 2007, 28(1): 71-74.

Zhang Guomin;Qiao Yuanyuan;Chen Yuping;Lü Zhe;An Yunqing. Cloning of Anti-keratin ScFv From a Human Large Phage Antibody Library[J]. Journal of Capital Medical University, 2007, 28(1): 71-74.

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