Construction of scFv phage display library with hapten-specific repertories and characterization of anti-ivermectin fragment isolated from the library

文献类型： 外文期刊

第一作者： Zhang, Xiao

作者： Zhang, Xiao;Zhang, Cunzheng;Liu, Yuan;Yu, Xiangyang;Liu, Xianjin

作者机构：

关键词： Phage display library;scFv;16-membered macrocyclic backbone micro-molecular substances;Screening

期刊名称：EUROPEAN FOOD RESEARCH AND TECHNOLOGY （ 影响因子：2.998； 五年影响因子：3.005 ）

ISSN： 1438-2377

年卷期： 2010 年 231 卷 3 期

页码：

收录情况： SCI

摘要： A specific phage display library was developed for screening antibodies against micro-molecular substances with 16-membered macrocyclic backbone. Through mRNA extraction, RT-PCR and gene splicing by overlap extension PCR (SOE-PCR), single-chain fragment variable (scFv) gene fragments about 750 bp were generated and ligated with the phagemid pCANTAB5E and were transformed into competent cells. A phage display library containing 2.4 x 10(6) clones was constructed from milbemycin oxime-bovine serum albumin (MILO-BSA) immunized mice. The screening was carried out by ivermectin-bovine serum albumin (IVM-BSA) with different concentration levels. Furthermore, scFv phage clones were isolated within the four round library panning and screening by phage- ELISA, 10 positive clones were obtained finally. These positive clones were then sequenced, and their soluble type antibodies were identified and showed significant binding activity.

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