抗甲基嘧啶磷单克隆抗体的研制及其在间接竞争酶联免疫吸附测定中的应用。

Development of monoclonal antibodies against pirimiphos-methyl and their application to IC-ELISA.

作者信息

Yang Zheng-You, Kolosova Anna Yu, Shim Won-Bo, Chung Duck-Hwa

机构信息

Division of Applied Life Science, Graduate School of Gyeongsan National University, Chinju, Gyeongnam 660-701, South Korea.

出版信息

J Agric Food Chem. 2006 Jun 28;54(13):4551-6. doi: 10.1021/jf0606196.

DOI:10.1021/jf0606196

PMID:16786997

Abstract

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%.

摘要

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%。