Bactrian camel nanobody-based immunoassay for specific and sensitive detection of Cry1Fa toxin

文献类型： 外文期刊

第一作者： Wang, Pingyan

作者： Wang, Pingyan;Li, Guanghui;Yan, Junrong;Wan, Yakun;Wan, Yakun;Hu, Yonghong;Zhang, Cunzheng;Liu, Xianjin

作者机构：

关键词： Nanobody;Phage-displayed library;Cry1Fa;Biotin;Streptavidin;Sandwich immune ELISA

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

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收录情况： SCI

摘要： The variable domain of the heavy-chain-only antibody (VHH) or nanobody (Nb), derived from camelids, begins to play an important role on the detection of protein markers. In this study, we constructed a phage-displayed library of VHHs against Cry1Fa by immunizing a healthy Bactrian camel with Cry1Fa toxin. After a series of bio-panning and screening by phage display technology, three anti-Cry1Fa nanobodies (Nbs) with great difference in complementarity determining region 3 (CDR3) were obtained and they were highly specific to Cry1Fa as well as showed full of activity when exposed to 70 degrees C for 3 h. Through modifying Nbs with Horseradish Peroxidase (HRP) and biotin, two Nbs which can recognize the different epitopes of Cry1Fa were determined and they were used to establish a novel sandwich immune ELISA based on biotin-SA interaction for Cry1Fa detection. The immunoassay exhibited a linear range from 1 to 100 ng/mL with a detection limit of 0.88 ng/mL. The recoveries from spiked corn and soybean samples were ranged from 83.33 to 117.17%, with a coefficient of variation (C.V) less than 6.0%. All together, the proposed immunoassay will be a promising way for sensitive and accurate determination of Cry1Fa toxin. (C) 2014 Elsevier Ltd. All rights reserved.

分类号： R9

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