Abstract:

In this study, artificial antigens for organophosphate insecticide triazophos including one immunogen and two coating antigens were prepared. The hapten (TZPM-Hap) of organophosphate insecticide triazophos was coupled to bovine serum albumin (BSA) by the active ester method to prepare the artificial immunogen TZPM-A-BSA. Coupling of TZPM-Hap to ovalbumin (OVA) by the active ester method or the mixed anhydride method produced the two coating antigens TZPM-A-OVA and TZPM-M-OVA, respectively. Balb/c mice were immunized with the immunogen, and the splenocytes of the immunized mice were fused with Sp2/0 cells to generate monoclonal antibody (McAbs). A hybridoma cell line (FC3) secreting antitriazophos McAbs was obtained, whose subtype was IgM. Compared with TZPM-M-OVA, better sensitivity in the icELISA format using TZPM-A-OVA was found, which demonstrated 9.7μg/L of IC50, 1.0μg/L of LOD, and 2.5－100.0μg/L of work range (IC20－IC80). The McAbs showed no obvious cross-reactivity with other structural-related organophosphate pesticides. The result indicated that one hybridoma cell line with high specificity and sensitivity was obtained, which will be of importance to develop an ELISA kit for triazophos in the future.

Key words: triazophos, monoclonal antibody, enzyme-linked immunosorbnent assay