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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

作者： Zhang, Xiao 1 ; Zhang, Cunzheng 1 ; Liu, Yuan 1 ; Yu, Xiangyang 1 ; Liu, Xianjin 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Qual Safety & Detect Res, Nanjing 210014, Peoples R China

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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[4]Established a new double antibodies Sandwich enzyme-linked immunosorbent assay for detecting Bacillus thuringiensis (Bt) Cry1Ab toxin based single-chain variable fragments from a naive mouse phage displayed library. Zhang, Xiao,Xu, Chongxin,Zhang, Cunzheng,Liu, Yuan,Xie, Yajing,Liu, Xianjin. 2014

[5]Rapid isolation of single-chain antibodies from a human synthetic phage display library for detection of Bacillus thuringiensis (Bt) Cry1B toxin. Zhang, Xiao,Liu, Yuan,Zhang, Cunzheng,Wang, Yun,Liu, Xianjin,Zhang, Xiao,Wang, Yun,Xu, Chongxin. 2012

[6]Selection of a Single Chain Variable Fragment Antibody against Ivermectin from a Phage Displayed Library. Wen, Shuang,Zhang, Xiao,Liu, Yuan,Zhang, Qingqing,Liu, Xianjin,Wen, Shuang,Liang, Jiansheng.

[7]Isolation of single chain variable fragment (scFv) specific for Cry1C toxin from human single fold scFv libraries. Wang, Yun,Zhang, Xiao,Liu, Xianjin,Wang, Yun,Zhang, Xiao,Zhang, Cunzheng,Liu, Yuan,Liu, Xianjin.

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