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[目的]制备毒死蜱单克隆抗体,为建立毒死蜱残留检测方法奠定基础。[方法]以毒死蜱原药与3-巯基丙酸为起始原料,合成半抗原并对其进行结构鉴定。利用该半抗原与牛血清蛋白(BSA)和卵清蛋白(OVA)偶联制备毒死蜱人工免疫抗原和包被抗原,再取已免疫的经过筛选的Balb/c小鼠脾细胞与小鼠骨髓瘤细胞SP2/0融合,用间接ELISA和间接竞争ELISA方法对2株细胞进行鉴定,并测定与其他农药的交叉反应率。[结果]获得两株稳定分泌毒死蜱单克隆抗体的细胞株,其检测范围为0.04～0.42mg/L,半数抑制浓度为0.135mg/L,最低检测限为0.03mg/L,与杀螟硫磷、对硫磷和马拉硫磷几乎没有交叉反应。[结论]成功制备两株分泌毒死蜱抗体的单克隆细胞株,初步建立了毒死蜱间接ELISA检测方法。
[Objective] The research aimed to prepare the monoclonal antibody of chlorpyrifos and lay the foundation for the establishment of the detection method of chlorpyrifos residue. [Method] With the chlorpyrifos original drug and 3 - mercaptopropionic acid as starting materials, hapten was synthesized and its structure was identified. The hapten was used to conjugate bovine serum albumin (BSA) and ovalbumin (OVA) to prepare artificial immune antigen and coated antigen, and then the immunized selected Balb / c mouse spleen cells and mouse myeloma Cells SP2 / 0 fusion, indirect ELISA and indirect competitive ELISA method for the identification of two cells, and determine the cross-reaction rate with other pesticides. [Result] The two cell lines stably secreting monoclonal antibodies against chlorpyrifos were obtained. The detection range was 0.04-0.42 mg / L, the half inhibitory concentration was 0.135 mg / L, the minimum detectable limit was 0.03 mg / L, , Parathion and malathion almost no cross-reaction. [Conclusion] Two monoclonal cell lines secreting chlorpyrifos antibodies were successfully prepared, and an indirect ELISA method for the detection of chlorpyrifos was established.