文献类型：期刊文献

英文题名：Insights into the Binding Profile of Anti-chlorpyrifos Recombinant Antibodies: From Computational Simulation to Immunoassay Validation

作者：Zou, Rubing[1] Guo, Yuanhao[1,2] Wang, Yan[1] Lu, Xinying[1] Ma, Zhongjie[1] Shou, Linfei[3] Liu, Yihua[4] Zhu, Guonian[1] Guo, Yirong[1]

第一作者：Zou, Rubing

通信作者：Guo, YR[1]

机构：[1]Zhejiang Univ, Inst Pesticide & Environm Toxicol, Key Lab Biol Crop Pathogens & Insects Zhejiang Pro, Minist Agr,Key Lab Mol Biol Crop Pathogens & Insec, Yuhangtang Rd 866, Hangzhou 310058, Peoples R China;[2]Zhejiang Gongshang Univ, Qual & Safety Engn Inst Food & Drug, Hangzhou 310018, Peoples R China;[3]Zhejiang Prov Plant Protect Quarantine & Pesticide, Hangzhou 310004, Peoples R China;[4]Chinese Acad Forestry, Res Inst Subtrop Forestry, Hangzhou 311400, Peoples R China

年份：2023

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20233214485008);Scopus(收录号：2-s2.0-85166428527);WOS:【SCI-EXPANDED(收录号:WOS:001027362100001)】；

基金：This research was supported by the Department of Science and Technology in Zhejiang Province (grant no. 2023C02039) and the Department of Agriculture and Rural Affairs in Zhejiang Province (grant no. 2022SNJF012).

语种：英文

外文关键词：Antibodies - Binding energy - Mammals - Molecular dynamics - Molecular modeling - Pesticides

摘要：A novelvirtual screening strategy was proposed for theprofilingand discovery of active variable regions (VRs) that encode hapten-specificrecombinant antibodies (rAbs). Chlorpyrifos, a hazardous organophosphoruspesticide, was selected as the target. First, a VR model-14G4 fromanti-chlorpyrifos hybridoma was built via homology modeling. Its bindingpattern toward seven organophosphorus analogues was assessed throughvirtual screening by performing molecular docking. Based on energyscoring, visual examination, and molecular interaction analysis, chlorpyrifos-methylwas also inferred as the high-affinity target for model-14G4 and wasthen confirmed via a non-competitive surface plasmon resonance (SPR)assay. Subsequently, we attempted to discover hapten-specific VRsby creating a collection of VR models for anonymous testing. Chlorpyrifosand model-14G4 were employed as the known hit and active VRs, respectively.After molecular docking, a novel anti-chlorpyrifos VR (model-1) wasidentified due to its satisfactory energy scoring and a similar bindingpattern to the reference model-14G4. Expressed by HEK293(F) mammaliancells, the newly prepared full-length rAb-model-1 and rAb-14G4 exhibitedhigh sensitivities for detecting chlorpyrifos by the indirect competitiveenzyme-linked immunosorbent assay (ic-ELISA), with IC50 of 3.01 ng/mL and 42.82 ng/mL, respectively. They recognized chlorpyrifos-methylwith a cross-reactivity (CR) of 2.5-17.3%. Moreover, the bindingproperties of rAb-model-1 for recognizing chlorpyrifos and chlorpyrifos-methylwere confirmed via a non-competitive microscale thermophoresis (MST)method. Thus, the experimental results showed good agreement withcomputational outputs on antibody profiling. Furthermore, the recognitiondiversity of rAb-model-1 for chlorpyrifos and chlorpyrifos-methylwas studied via molecular dynamics simulation. Overall, the proposedstudy provides a versatile and economical strategy for antibody characterizationand promotes the in vitro production of rAbs forpesticide monitoring.