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基于单克隆抗体的酶联免疫吸附法检测拟除虫菊酯类杀虫剂溴氰菊酯的研究。

Development of a monoclonal antibody-based enzyme immunoassay for the pyrethroid insecticide deltamethrin.

机构信息

Chinese Academy of Agricultural Sciences, Wuhan 430062, China.

出版信息

J Agric Food Chem. 2010 Jul 28;58(14):8189-95. doi: 10.1021/jf101483w.

Abstract

With a screening hemisolid stem-cell culture, four positive hybridomas 2B(12), 2C(1), 2F(1), and 3D(4) were screened and used to prepare four correspondent monoclonal antibodies (McAbs) against the pyrethroid insecticide deltamethrin. These McAbs showed I(50) values in a range of 17-94 ng mL(-1), from among which the antibody 2B(12) with the lowest I(50) value was selected to develop an optimized enzyme-linked immunosorbent assay (ELISA). In the developed ELISA, the I(50) of deltamethrin was 17.0 +/- 3.3 ng mL(-1) and the limit of detection (LOD) was 1.2 +/- 1.3 ng mL(-1). There seemed to be little or no cross-reactivity with other tested pyrethroids and their metabolites. For validation of the assay method, environmental water samples fortified with deltamethrin were analyzed with the ELISA and gas chromatography (GC) methods. The recoveries of the developed ELISA ranged from 82 to 117%, which were close to those of the GC method (94-103%). These results suggested that the developed ELISA based on the McAb 2B(12) could be used for the rapid and sensitive determination of deltamethrin in environmental water.

摘要

采用半固体干细胞筛选法,筛选出了 4 株阳性杂交瘤细胞 2B(12)、2C(1)、2F(1)和 3D(4),并以此制备了 4 种针对拟除虫菊酯类杀虫剂溴氰菊酯的单克隆抗体(McAbs)。这 4 种 McAbs 的半数抑制浓度(I(50))在 17-94ng/mL 之间,其中 I(50)值最低的抗体 2B(12)被选中用于优化酶联免疫吸附测定(ELISA)。在优化后的 ELISA 中,溴氰菊酯的 I(50)值为 17.0 ± 3.3ng/mL,检测限(LOD)为 1.2 ± 1.3ng/mL。该方法与其他测试的拟除虫菊酯及其代谢物的交叉反应性很小或没有。为了验证该检测方法,采用 ELISA 和气相色谱(GC)法对加标环境水样进行了分析。该 ELISA 的回收率在 82%至 117%之间,与 GC 法(94%-103%)接近。这些结果表明,基于 McAb 2B(12)的 ELISA 可用于快速、灵敏地检测环境水中的溴氰菊酯。

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