Production of Broad Specificity Anti-Methoxyorganophosphorus Pesticides Antibody
The O,O-dimethyl phosphorodithioate,which is the generic structure of methoxy organophosphorus pesticide,was used as the target determinate group to generate broad specificity polycolonal antibodies against a group of organophosphorus pesticides.S-carboxymethyl O,O-dimethylphorodithioate(CMP) was synthesized as generic hapten and conjugated with the carrier proteins bovine serum albumin(BSA) and ovalbumin(OVA) by the mixed anhydride(MA) and active ester(AE) method respectively.New Zealand white rabbits were immunized with CMP-MA-BSA and CMP-AE-BSA to produce the polyclonal antibodies(PAbs).The PAbs displayed high level for CMP-AE-BSA with the favorable titers of 256000.After optimization of the ELISA conditions such as organic solvents and pH values,an indirect ELISA procedure for the determination of CMP was established.Based on statistical analysis,the limit deteciton was 0.076 μg/L and IC50 was 93.97 μg/L.Different cross-reactions with 14 methoxy organophosphorus pesticide including malathion,phenthoate,dimethoate,phosmet,fenthion,pirimiphos-methyl,methyl parathion,fenitrothion,methidathion were observed,with IC50 value of 69.92,136.90,230.39,416.84,508.57,510.38,607.21,835.30 and 850.21 μg/L respectively.The result indicated the antibody was practicable screening tool for quantities and semi-quantities detection of the nine methoxy organophosphorus pesticides.