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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

文献类型： 外文期刊

作者： Zhang, Xiao 1 ; Xu, Chongxin 1 ; Zhang, Cunzheng 1 ; Liu, Yuan 1 ; Xie, Yajing 1 ; Liu, Xianjin 2 ;

作者机构： 1.Jiangsu Acad Agr Sci, Key Lab Food Qual & Safety Jiangsu Prov, Key Lab Control Technol & Stand Agroprod Safety &, Minist Agr,Inst Food Qual Safety & Detect Res,Sta, Nanjing 210014, Peoples R China

2.Jiangsu Acad Agr Sci, Key Lab Food

关键词： Cry1Ab toxin;ScFv;Naive mouse phage displayed library;Double antibodies sandwich immunoassay

期刊名称：TOXICON （ 影响因子：3.033； 五年影响因子：2.689 ）

ISSN： 0041-0101

年卷期： 2014 年 81 卷

页码：

收录情况： SCI

摘要： ScFvs are composed of the variable regions of the heavy and light chains via a short linker that maintain the specific antigen binding abilities of antibodies. In this study, we constructed a naive mouse phage displayed library to generate scFvs against Cry1Ab toxin. After affinity panning, positive phage-scFvs were isolated, sequenced and characterized by ELISA. The best binding ability scFv-G9 was expressed and purified. SDS-PAGE indicated that the relative molecular mass of scFv was estimated at 28 kDa. The purified scFv-G9 was used to develop a new DAS-ELISA for detecting Cry1Ab toxin, within minimum detection limit of 0.008 mu g mL(-1), a working range 0.018-6.23 mu g mL(-1), and the linear curve displayed an acceptable correlation coefficient of 0.98. The cross-reactivity showed that scFv-G9 had strongly binding ability to CrylAc toxin, but not to Cry1B, Cry1C and Cry1F toxin. The average recoveries of Cry1Ab toxin from spiked leaf and rice samples were in the range 92.1-94.8%, and 91.6-98.6%, respectively, with a coefficient of variation (C.V) less than 5.0%. These results showed promising applications of scfv-G9 for detecting Cry1Ab toxin with new DAS-ELISA. 2014 Elsevier Ltd. (C) All rights reserved.

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