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Construction and characterization of a class-specific single-chain variable fragment against pyrethroid metabolites

文献类型： 外文期刊

作者： Liu, Yuan 1 ; Liu, Dan 3 ; Shen, Chen 1 ; Dong, Sa 3 ; Hu, Xiaodan 1 ; Lin, Manman 1 ; Zhang, Xiao 1 ; Xu, Chongxin 1 ; Zhon 1 ;

作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Safety, Key Lab Food Qual & Safety Jiangsu Prov, Room 213, Nanjing 210014, Peoples R China

2.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Jiangsu, Peoples R China

3.Yangzhou Univ, Sch Hort & Plant Protect, Yangzhou 225009, Jiangsu, Peoples R China

关键词： Single-chain variable fragment (scFv); 3-phenoxybenzoic acid (3-PBA); 3-phenoxybenzaldehyde (PBAld); 3-phenoxybenzyl alcohol (PBAlc)

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.813； 五年影响因子：4.697 ）

ISSN： 0175-7598

年卷期： 2020 年 104 卷 17 期

页码：

收录情况： SCI

摘要： Pyrethroids are insecticides that are widely used in rural and urban areas worldwide. After entering the environment, pyrethroids are rapidly metabolized or degraded by various biological or abiotic methods. In this study, a single-chain variable fragment (scFv) which could simultaneously detect three pyrethroid metabolites was constructed based on a hybridoma raised against 3-phenoxybenzoic acid (3-PBA). By molecular docking, it showed that there were hydrogen bonds, hydrophobic interactions, CH-pi interaction, and cation-pi interaction between 3-PBA and its scFv. All the contact residues contributing to hydrogen bonds are located in VH-CDR2 or its neighboring region, and two of them were mutants of the closest germline sequence. Based on competitive ELISA, the half maximal inhibitory concentration (IC50) of the scFv for 3-PBA, 3-phenoxybenzaldehyde (PBAld), and 3-phenoxybenzyl alcohol (PBAlc) were calculated to be 0.55, 0.59, and 0.63 mu gmL(-1), respectively. The scFv also showed 23.91%, 13.41%, 1.15%, 1.00%, and 0.56% cross-reactivity with phenothrin, deltamethrin, fenvalerate, beta-cypermethrin, and fenpropathrin. The broad specificity of the scFv may be due to its hapten design. The scFv could be employed in class-specific immunoassays for pyrethroid metabolites with phenoxybenzyl (PB) group. It is also potentially used for characterizing degradation of pyrethroids or detecting PBAlc (PBAld) alone, and the detection results should be confirmed by other selective methods.

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