Abstract:
Objective To produce monoclonal antibodies (McAb) against chlorpyrifos and to establish a rapid assay for detecting the residues of chlorpyrifos.
Methods Hapten of chlorpyrifos was synthesized by technical grade imidacloprid reacted with 3-mercaptopropionic acid and the structure of synthesized product was identified.Then,the hapten was covalently conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to prepare immunogen and coating antigen.Two hybridoma cell lines secreting McAb against chlorpyrifos were 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 enzyme-linked immunosorbent assay (ELISA) and indirect competitive ELISA; cross-reactivity with other organophosphate pesticides was also tested.
Results Two monoclonal cell lines that secreted Chlorpyrifos antibodies steadily were obtained.Based on the calibration curve,the range of detection was 0.04-0.42 mg/L,the median inhibitory concentration was 0.135 mg/L,and the limit of detection was 0.03 mg/L.No cross-reactivity of the antibody with sumithion,parathion and malathion was observed.
Conclusion Two chlorpyrifos monoclonal antibody cell lines are prepared successfully.A ELISA is preliminarily developed using the monoclonal antibody to detect chlorpyrifos.
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