[关键词]
[摘要]
本文以RAC-linker-BSA为免疫抗原获得了莱克多巴胺的多克隆抗体,用混合酸酐法合成包被抗原,建立了莱克多巴胺的间接竞争酶联免疫检测方法(ELISA),并对抗体性质进行了评价。结果表明,制备出的莱克多巴胺抗体具有较高的灵敏度,IC50为0.9 ng/mL,检出限(IC15)为0.1 ng/mL,低于同类文献的报道结果;除了与多巴酚丁胺的交叉反应为7.5%外,与克伦特罗、沙丁胺醇、特步他林、异丙肾上腺素和异奎胍均无交叉现象,说明抗体特异性较高;此外,抗体具有很好的稳定性,可在4 ℃冰箱中储存半年以上。为建立酶联免疫快速检测方法,实现对莱克多巴胺的有效监控奠定了良好的基础。
[Key word]
[Abstract]
An indirect competitive enzyme-linked immunosorbent assay (ELISA) procedure was established to detect ractopamine employing a polyclonal antibody generated from RAC-linker-BSA and a coating antigen synthesized by method of mixed acid anhydride. The antibody against ractopamine showed high sensitivity with an IC50 of 0.9 ng/mL and the detection limit (IC15) was 0.1 ng/mL, which was lower than the data reported in literatures. The antibody with high specificity showed no cross-reactivity with clenbuterol, salbutamol, isoproterenol, terbutaline and isoxsuprine, while had the cross-reactivity of 7.5% with dobutamine. In addition, it was stable enough to be stored at 4 ℃ for half a year. This study laid a well foundation for the control of ractopamine by ELISA method.
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[基金项目]
国家十一五科技支撑计划(2006BAD05A06)