Yang Zheng-You, Kolosova Anna Yu, Shim Won-Bo, Chung Duck-Hwa
Division of Applied Life Science, Graduate School of Gyeongsan National University, Chinju, Gyeongnam 660-701, South Korea.
J Agric Food Chem. 2006 Jun 28;54(13):4551-6. doi: 10.1021/jf0606196.
To detect the organophosphorus (OP) pesticide pirimiphos-methyl in grain samples, a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed and optimized. By the active esters method, pirimiphos-methyl hapten A was conjugated to keyhole limpet hemocyanin to be used as the immunogen for the production of monoclonal antibodies, and pirimiphos-methyl hapten B was conjugated to ovalbumin to be used as coating antigen. By using the monoclonal antibody and the coating antigen, an IC-ELISA has been developed. Under the established optimized conditions, the IC-ELISA showed an IC50 of 4.2 ng/mL with a detection limit of 0.07 ng/mL. The IC-ELISA showed negligible cross-reactivity with other OP pesticides except with pirimiphos-ethyl. Recoveries of pirimiphos-methyl from spiked grain samples ranged from 83 to 96%.
为检测谷物样品中的有机磷（OP）农药甲基嘧啶磷，开发并优化了一种基于单克隆抗体的间接竞争酶联免疫吸附测定法（IC-ELISA）。通过活性酯法，将甲基嘧啶磷半抗原A与钥孔血蓝蛋白偶联用作制备单克隆抗体的免疫原，将甲基嘧啶磷半抗原B与卵清蛋白偶联用作包被抗原。利用该单克隆抗体和包被抗原，建立了IC-ELISA。在既定的优化条件下，该IC-ELISA的半数抑制浓度（IC50）为4.2 ng/mL，检测限为0.07 ng/mL。除了与乙基嘧啶磷有交叉反应外，该IC-ELISA与其他OP农药的交叉反应可忽略不计。加标谷物样品中甲基嘧啶磷的回收率为83%至96%。
