Production and characterization of monoclonal antibody for class-specific determination of O,O-dimethyl organophosphorus pesticides and effect of heterologous coating antigens on immunoassay sensitivity
文献类型: 外文期刊
第一作者: Liu, Yan
作者: Liu, Yan;Lou, Yang;Xu, Dan;Qian, Guoliang;Wu, Runru;Hu, Baishi;Liu, Fengquan;Zhang, Qi
作者机构:
关键词: Organophosphorus pesticides;Monoclonal antibody;Class-specific manner;Heterology;ELISA;Sensitivity
期刊名称:MICROCHEMICAL JOURNAL ( 影响因子:4.821; 五年影响因子:4.364 )
ISSN:
年卷期:
页码:
收录情况: SCI
摘要: A generic hapten (H1), 3-(4-Dimethoxyphosphorothioyloxy phenyl)propanoic acid, was synthesized to produce monoclonal antibodies (mAbs) for the determination of organophosphorus pesticides (OPs) in a class-specific manner. Six heterologous haptens were designed to study the effect of hapten heterology on immunoassay sensitivity. Several mice were immunized with this H1-BSA immunogen. Spleen cells of two immunized mice were fused with myeloma cells, and the resulting hybridomas were screened using HI-OVA. In a competitive indirect enzyme-linked immunosorbent assay (ELISA) format, three hybridoma cell lines (B4-C6, D12-B5, E5-H2) that produced mAbs with high selectivity and broad specificity were selected and expanded. The monoclonal antibody D12-B5 with higher titer was chosen for further study. In heterologous assay, the combination of D12-B5 and coating antigen H7-OVA constituted a particularly sensitive assay for competitive indirect ELISA and showed broad specificity for the determination of OPs, including parathion-methyl, chlorpyrifos-methyl, tolclofos-methyl. fenthion, malathion, fenitrothion. This combination resulted in the IC50 of 0.58-10.47 mu g/ml.
分类号: O655.11
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