文献类型：期刊文献

英文题名：Computer-aided profiling of a unique broad-specific antibody and its application to an ultrasensitive fluoroimmunoassay for five N-methyl carbamate pesticides

作者：Zou, Rubing[1,2] Guo, Yuanhao[1] Chen, Yang[1] Zhao, Ying[1] Zhao, Li[3] Zhu, Guonian[1] Liu, Yihua[4] Peters, Jeroen[2] Guo, Yirong[1]

第一作者：Zou, Rubing

通信作者：Guo, YR[1]

机构：[1]Zhejiang Univ, Key Lab Mol Biol Crop Pathogens & Insects, Minist Agr, Key Lab Biol Crop Pathogens & Insects Zhejiang Pr, Hangzhou 310058, Peoples R China;[2]Wageningen Food Safety Res, Akkermaalsbos 2, NL-6708 WB Wageningen, Netherlands;[3]Shanghai Agr Technol Extens Serv Ctr, Shanghai 201103, Peoples R China;[4]Chinese Acad Forestry, Res Inst Subtrop Forestry, Hangzhou 311400, Peoples R China

年份：2022

卷号：426

外文期刊名：JOURNAL OF HAZARDOUS MATERIALS

收录：;EI(收录号：20214911292837);Scopus(收录号：2-s2.0-85120638730);WOS:【SCI-EXPANDED(收录号:WOS:000752577200002)】；

基金：This work was supported by grants from Shanghai Agriculture Applied Technology Development Program, China (No. 2020-02-08-00-08-F01456) and Key Research & Development Program in Zhejiang Province, China (No. 2020C02024-2) .

语种：英文

外文关键词：molecular docking; virtual screening; broad-specific monoclonal antibody; microsphere-based immunoassay; N-methyl carbamate

摘要：Pollution of N-methyl carbamate (NMC) pesticides is threatening the non-target organisms' survival. Thus, broad-specific antibodies and class-selective immunoassays are demanding for multiple NMCs determination. In this study, we employed a molecular docking-based virtual screening strategy to fast profile antibody spectrum, based on a designed chemical pool containing 17 compounds. A monoclonal antibody (mAb)-6G against carbofuran was used as the objective. The recombinant full-length IgG was successfully expressed to validate the antibody sequences for homology modeling. After docking, we manually categorized the antibody-chemical binding strength into three groups. Non-competitive surface plasmon resonance (SPR) demonstrated the mAb6G affinitive binding toward five NMCs (carbofuran, isoprocarb, propoxur, carbaryl and carbosulfan), which were classified into strong and moderate binding categories. Antibody binding properties were confirmed again by ic-ELISA and lateral flow immunochmmatographic strip. Subsequently, an ultrasensitive indirect competitive fluommicrosphere-based immunoassay (ic-FMIA) was established with the IC50 (half-maximal inhibitory concentration) values of 0.08-3.37 ng/mL. This portable assay presented a 30-230-fold improved sensitivity than traditional ic-ELISA and was applied in European surface water analysis. Overall, our work provides an efficient platform integrating in-silico and experimental methodologies to accelerate the characterization of hapten-specific antibody binding properties and the development of high-sensitive immunoassays for multi-pollutants monitoring.