Development and comparative study of chemosynthesized antigen and mimotope-based immunoassays for class-specific analysis of O,O-dimethyl organophosphorus pesticides

Abstract

The multi-residue determination of organophosphorus pesticides (OPs) is an important task due to the wide application and high toxicity of OPs. However, there is no promising immunoassay to monitor the multi-residue of O,O-dimethyl OPs. In this study, a monoclonal antibody (mAb) against a generic hapten of O,O-dimethyl OPs (O,O-dimethyl O-(3-carboxyphenyl)phosphorothioate) was prepared. To develop an effective class-specific immunoassay, two strategies were performed to select the appropriate coating antigen or competing antigen. On the one hand, a total of 20 haptens were chemosynthesized, attached to ovalbumin for use as coating antigen candidates, and selected by direct competitive ELISA (dcELISA). As a second strategy, mimotopes of the mAb were selected from a random phage-display peptide library by panning, and the optimum mimotope was expressed as a fusion protein and biotinylated in vitro. Based on the selected chemosynthesized coating antigen and the biotinylated mimotope fusion protein, two sensitive broad-specificity dcELISAs were developed. The sensitivity, selectivity and practicability of the two immunoassays were compared. The results demonstrated that both methods showed similar selectivity and sensitivity and were reliable for O,O-dimethyl OP residues screening. However, the screening operation of mimotopes was much simpler and safer compared to the preparation of chemosynthesized coating antigens.