Preparation of monoclonal antibody against chlorprifos and development of ELISA for detection of chlorprifos residues

• 1、Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University

Abstract：Objective To produce monoclonal antibodies (McAb) against chlorpyrifos and to establish rapid assay for the resides of chlorprifos. Methods Hapten of chlorprifos was synthesized by using technical grade imidacloprid reacted with 3-Mercaptopropionic acid and the structure was identified. The hapten was covalently conjugated to BSA and OVA to prepare immunogen and coating antigen. Two hybridoma cell lines secreting monoclonal antibodies (McAb) against chlorpyrifos had been established by fusing mouse myeloma cells SP2/0 and splenocytes from the Balb/c mouse immunized with immunogen. Two cell lines were identified by indirect ELISA and indirect competitive ELISA; Cross-reactivity with other pesticides was also determined. Results Two monoclonal cell lines that secreted chlorprifos antibodies steadily were obtained . Based on the calibration curve, the linear detection was 0.04～0.42μg/mL and the IC50 was 0.135μg/mL. No cross-reactivity of the antibody with Sumithion parathion and malathion was observed, indicating that the antibody is highly specific for chlorprifos. Conclusions Two chlorpyrifos monoclonal antibody cell lines were prepared successfully. A ELISA was preliminarily developed using the monoclonal antibody to detect chlorprifos.

Keywords： chlorprifos artificial antigen monaclonal antibody enzyme-linked immunosorbent assay (ELISA)