Production of Antigen-Binding Fragment against O,O-Diethyl Organophosphorus Pesticides and Molecular Dynamics Simulations of Antibody Recognition

Chen, Zijian; Zhang, Xuan; Wang, Bingfeng; Rao, Meifang; Wang, Hong; Lei, Hongtao; Liu, Hui; Zhang, Yan; Sun, Yuanming; Xu, Zhen-Lin

doi:10.3390/ijms19051381

Cited by 11 publications

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References 42 publications

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“…Compared with the reported anti-chlorpyrifos mAbs, − scFvs , and Fab (IC 50 : 2.08–1600 ng/mL), our newly prepared mAb-model-1 and rAb-model-1 achieved satisfying sensitivities and limit of detections (LODs) (0.17–1.11 ng/mL) (Table S5). Moreover, they exhibited fewer CRs (CR ≤ 3.6%) to chlorpyrifos-methyl, in contrast to mAb-14G4, rAb-14G4, and previously reported anti-chlorpyrifos antibodies with similar IC 50 values for chlorpyrifos. , As Table S6 depicts, model-1 and model-14G4 showed identities of 42.2 and 84.1% for VH and VL, with similarities of 65.5 and 93.5%, respectively.…”

Section: Resultsmentioning

confidence: 83%

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

et al. 2023

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A novel virtual screening strategy was proposed for the profiling and discovery of active variable regions (VRs) that encode hapten-specific recombinant antibodies (rAbs). Chlorpyrifos, a hazardous organophosphorus pesticide, was selected as the target. First, a VR model-14G4 from anti-chlorpyrifos hybridoma was built via homology modeling. Its binding pattern toward seven organophosphorus analogues was assessed through virtual screening by performing molecular docking. Based on energy scoring, visual examination, and molecular interaction analysis, chlorpyrifos-methyl was also inferred as the high-affinity target for model-14G4 and was then confirmed via a non-competitive surface plasmon resonance (SPR) assay. Subsequently, we attempted to discover hapten-specific VRs by creating a collection of VR models for anonymous testing. Chlorpyrifos and model-14G4 were employed as the known hit and active VRs, respectively. After molecular docking, a novel anti-chlorpyrifos VR (model-1) was identified due to its satisfactory energy scoring and a similar binding pattern to the reference model-14G4. Expressed by HEK293(F) mammalian cells, the newly prepared full-length rAb-model-1 and rAb-14G4 exhibited high sensitivities for detecting chlorpyrifos by the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), with IC50 of 3.01 ng/mL and 42.82 ng/mL, respectively. They recognized chlorpyrifos-methyl with a cross-reactivity (CR) of 2.5–17.3%. Moreover, the binding properties of rAb-model-1 for recognizing chlorpyrifos and chlorpyrifos-methyl were confirmed via a non-competitive microscale thermophoresis (MST) method. Thus, the experimental results showed good agreement with computational outputs on antibody profiling. Furthermore, the recognition diversity of rAb-model-1 for chlorpyrifos and chlorpyrifos-methyl was studied via molecular dynamics simulation. Overall, the proposed study provides a versatile and economical strategy for antibody characterization and promotes the in vitro production of rAbs for pesticide monitoring.

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