中文摘要：转基因作物引起的环境安全问题和人类健康的潜在风险问题备受关注。最近的研究表明，源自农杆菌属菌株CP4蛋白（CP4-EPSPS）的5-烯醇基丙基莽草酸-3-磷酸合成酶（EPSPS）对草甘膦除草剂具有很高的抗性，是转基因作物的典型生物标志物。鉴由此，在作物中能检测出CP4-EPSPS蛋白的灵敏及方便的策略很受人们期待。本文基于偶合纳米体、有序介孔碳（OMC）和硫堇（TH）研发了一个简单的电化学免疫传感器。研究发现，采用差分脉冲伏安法（DPV）可以高效、高效地测定CP4-EPSPS蛋白，表明所提出的电化学免疫传感器在CP4-EPSPS的筛选和检测中具有巨大的潜力。
外文摘要：The problems of environmental security and the potential risks of human health caused by transgenic crops have attracted much attention. Recent studies reveal 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium sp. strain CP4 protein (CP4-EPSPS), which shows very high resistance to herbicide glyphosate, is a typical biomarker of genetically modified (GM) crops. For this reason, it is highly anticipated to devise a sensitive and convenient strategy to detect CP4-EPSPS protein in crops. Herein, we report a simple electrochemical immunosensor by coupling nanobody, ordered mesoporous carbon (OMC), and thionine (Th). As a capture agent, the nanobody was screened out from an immunized Bactrian camel, and exhibited superior properties with respect to conventional antibody, such as higher stability and stronger heat resistance. Moreover, OMC offered an effective platform with high surface area, electrical conductivity, and biocompatibility, which greatly facilitated the assembly of redox probe Th, and further coupling of large amount of capture nanobodies. As a result, the CP4-EPSPS protein could be determined with high sensitivity and efficiency by differential pulse voltammetry (DPV) in a wide linear range from 0.001 to 100 ng.mL(-1) with a low detection limit of 0.72 pg.mL(-1), which was more than 3 orders of magnitude lower than those of previously reported works. As an example, the proposed electrochemical immunosensor was successfully applied to spiked samples, demonstrating its great potential in CP4-EPSPS screening and detection.
外文关键词：electrochemical immunoassay, mesoporous carbon, nanobody, genetically modified crops, CP4-EPSPS
作者：Zhang, Mingming; Li, Guanghui; Zhou, Qing; Pan, Deng; Zhu, Min; Xiao, Runyu; Zhang, Yuanjian; Wu, Guoqiu; Wan, Yakun; Shen, Yanfei
作者单位：Southeast University
期刊名称：ACS SENSORS
期刊影响因子：0.0
出版年份：2018
出版刊次：5
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