Simultaneous production of monoclonal antibodies against Bacillus thuringiensis (Bt) Cry1 toxins using a mixture immunization
Simultaneous production of monoclonal antibodies against Bacillus thuringiensis (Bt) Cry1 toxins using a mixture immunization
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使用混合免疫同时生产抗苏云金芽孢杆菌 (Bt) Cry1 毒素的单克隆抗体
DOI：
10.1016/j.ab.2017.05.016
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发表时间：
2017-08-15
影响因子：
2.9
通讯作者：
Liu, Xianjin
中科院分区：
文献类型：
--
作者：
Dong, Sa;Zhang, Cunzheng;Liu, Xianjin
Cry1毒素的检测主要依赖于基于特异性单抗的免疫分析。在本研究中，用7种Cry1毒素进行了混合免疫。结果表明，获得了5株不同特性的单抗，其中1株能识别所有7种Cry1毒素的单抗命名为5-E8。以5-E8单抗为基础，建立了检测7种Cry1毒素的双抗体夹心酶联免疫吸附试验(DAS-ELISA)，检测限为6.37~11.35 ng mL(-1)，定量限为17.36~24.48 ng mL(-1)，与Cry2A和vip3无交叉反应。回收率试验表明，在定量范围内(LOQ-100ngmL(-1))，回收率为78%~110%。建立的DAS-ELISA法可作为农产品中Cry1毒素监测的有效工具。混合免疫为在一个免疫周期内同时产生多种单抗开辟了新的途径。(C)2017 Elsevier Inc.保留所有权利。
The detections of Cry1 toxins are mainly dependent on immunoassays based on specific monoclonal antibodies (mAb). In the present study, a mixture immunization with seven Cry1 toxins was administered. The results showed that five mAbs with different characteristics, especially one mAb named 5-E8 which could recognize all the seven Cry1 toxins were obtained. Based on the 5-E8 mAb, a double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) which can specifically detect the seven Cry1 toxins without cross-reactivity to Cry2A and vip3 was developed with the limit of detection (LOD) and limit of quantification (LOQ) of 6.37-11.35 ng mL(-1) and 17.36-24.48 ng mL(-1), respectively. The recovery tests showed that the recoveries ranged from 78% to 110% within the quantitation range (LOQ-100 ng mL(-1)). The established DAS-ELISA can be a useful tool for monitoring the Cry1 toxins in agricultural products. Mixture immunization opens a new path for producing diverse mAbs simultaneously in a single immunization circle. (C) 2017 Elsevier Inc. All rights reserved.