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设计一种新型半抗原,并开发一种用于环境水样中麦草畏分析的灵敏单克隆免疫分析方法。

Design of a novel hapten and development of a sensitive monoclonal immunoassay for dicamba analysis in environmental water samples.

机构信息

Department of Organic Chemistry, University of Valencia, Doctor Moliner 50, Burjassot 46100, València, Spain.

Institute of Agricultural Chemistry and Food Technology (IATA), Spanish Council for Scientific Research (CSIC), Av. Agustí Escardino 7, Paterna 46980, València, Spain.

出版信息

Sci Total Environ. 2022 Nov 20;848:157770. doi: 10.1016/j.scitotenv.2022.157770. Epub 2022 Aug 1.

Abstract

Weed resistance to glyphosate has been a driving force behind the increased use of alternative herbicides in agriculture. Recently, dicamba-tolerant recombinant plants were introduced to the market, which may result in residues of this agrochemical contaminating environmental waters. Given that restrictions on the use of dicamba have consequently been established by regulatory agencies, it is therefore also desirable to conduct extensive controls on dicamba residues. Immunoassays are currently the most powerful bioanalytical technology for the rapid monitoring of chemical residues and contaminants. In the present study, a novel hapten was designed maintaining unaltered all the antigenic moieties of the target molecule, and this was used to generate high-affinity monoclonal antibodies against dicamba for the first time. Additionally, a collection of haptens with different linker composition or linker tethering site was synthesized and conjugated to proteins. Using these novel immunoreagents, a direct competitive enzyme-linked immunosorbent assay with a limit of detection for dicamba of 0.24 ng/mL was developed and validated. Analysis of water samples from different origins afforded recovery values between 90 % and 120 %, and coefficients of variation below 20 % were obtained. These results indicate that the developed immunochemical assay is suitable for the rapid determination of dicamba residues in environmental water samples.

摘要

草甘膦抗性杂草已成为农业中增加使用替代除草剂的主要驱动力。最近,耐草铵膦的重组植物被推向市场,这可能导致这种农用化学品的残留污染环境水。鉴于监管机构随后对草铵膦的使用制定了限制,因此也需要对草铵膦残留进行广泛的控制。免疫测定法目前是快速监测化学残留物和污染物的最强大的生物分析技术。在本研究中,设计了一种新型半抗原,保持了目标分子所有抗原部分的不变,并首次用于生成针对草铵膦的高亲和力单克隆抗体。此外,还合成了一系列具有不同连接子组成或连接子连接点的半抗原,并将其与蛋白质偶联。使用这些新型免疫试剂,开发并验证了一种用于检测草铵膦的直接竞争酶联免疫吸附测定法,其检测限为 0.24ng/mL。对来自不同来源的水样进行分析,得到的回收率在 90%至 120%之间,变异系数低于 20%。这些结果表明,所开发的免疫化学分析方法适用于快速测定环境水样中的草铵膦残留。

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