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

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2018 May 9

PMID 29734787

Citations 3

Authors

Zi-Jian Chen

Xuan Zhang

Bing-Feng Wang

Mei-Fang Rao

Hong Wang

Hong-Tao Lei

Hui Liu

Yan Zhang

Yuan-Ming Sun

Zhen-Lin Xu

Affiliations

Soon will be listed here.

Abstract

Immunoassay for pesticides is an emerging analytical method since it is rapid, efficient, sensitive, and inexpensive. In this study, a recombinant antigen-binding fragment (Fab) against a broad set of ,-diethyl organophosphorus pesticides (DOPs) was produced and characterized. The κ chain and Fd fragment were amplified via PCR and inserted into the vector pComb3XSS and the soluble Fab on phagemid pComb3XSS was induced by isopropyl -d-thiogalactoside in TOP 10F’. SDS-PAGE, Western blotting, and indirect competitive ELISA results indicated that Fab maintained the good characteristics of the parental mAb. To better understand antibody recognition, the three-dimensional (3D) model of Fab was built via homologous modeling and the interaction between Fab and DOPs was studied via molecular docking and dynamics simulations. The model clearly explained the interaction manner of Fab and DOPs, and showed that the Arg-L96 and Arg-H52 were mainly responsible for antibody binding. This work provided a foundation for further mutagenesis of Fab to improve its characteristics.

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