[噬菌体展示单链抗体片段表达载体的构建及小鼠未免疫抗体文库的构建]

[Construction of expressing vector for phage display scFv and a mouse unspecified antibody library].

作者信息

Zhang W G, Liu X F, Wang Y, Jian X, Lin Q, Chen A, Wu C W, Huang H M, Li Z H, Zhang Y, Huang H L

机构信息

Institute of Genetics, Chinese Academy of Sciences, Beijing.

出版信息

Yi Chuan Xue Bao. 1999;26(2):99-106.

PMID:10375858

Abstract

The display of antibody gene library on the surface of E. coli filamentous phage offers a new method of obtaining antibodies against specific antigens. In the paper, it is reported that a phagemid for phage display antibody, named after pFUW80, characterized expressing scFv either secretly or stickily. With a series of designed PCR primers, heavy-chain and light-chain variable region genes of mice antibodies were amplified and a mouse unspecified single-chain antibody library with the size of 1.2 x 10(6) clones was constructed. From this library, phage was selected out against antigen human IgG and detected by ELISA and partial sequence analysis. These primary results established the foundation for future research using this system.

摘要

将抗体基因文库展示于大肠杆菌丝状噬菌体表面，为获得针对特定抗原的抗体提供了一种新方法。本文报道了一种用于噬菌体展示抗体的噬菌粒，命名为pFUW80，其特点是能够以分泌或黏附的方式表达单链抗体（scFv）。利用一系列设计好的聚合酶链式反应（PCR）引物，扩增出小鼠抗体的重链和轻链可变区基因，并构建了一个库容为1.2×10⁶个克隆的小鼠未指定单链抗体文库。从该文库中筛选出针对人免疫球蛋白G（human IgG）抗原的噬菌体，并通过酶联免疫吸附测定（ELISA）和部分序列分析进行检测。这些初步结果为今后利用该系统开展研究奠定了基础。