| 莱克多巴胺免疫亲和柱的制备与应用研究 |
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| 引用本文： | 王迪,杨曙明,刘潇威,于洪侠.莱克多巴胺免疫亲和柱的制备与应用研究[J].分析测试学报,2010,29(8). |
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| 作者姓名： | 王迪 杨曙明 刘潇威 于洪侠 |
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| 作者单位： | 1. 农业部环境保护科研监测所,农业部/天津市产地环境与农产品安全重点开放实验室,天津,300191
2. 中国农业科学院农业质量标准与检测技术研究所,北京,100081 |
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| 基金项目： | 农业部948项目资助 |
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| 摘 要： | 用多元酸酐与混合酸酐相结合的方法合成了莱克多巴胺(Rac)抗原,免疫动物获得特异性抗体,并以蛋白A柱纯化得到IgG抗体。琼脂糖凝胶(Sepharose 4B)经溴化氰(CNBr)活化后与IgG抗体偶联,制备莱克多巴胺免疫吸附剂。据此建立了尿液中莱克多巴胺的免疫亲和柱净化/液相色谱-荧光法(HPLC-FL)测定的分析方法。免疫制备特异抗体50%抑制浓度(IC50)为5μg/L。Sepharose 4B经CNBr活化后与2 mg抗体的偶联率达87.4%。1 mL吸附剂的柱容量为67.57 ng。尿液中莱克多巴胺的回收率为76%~90%。
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| 关 键 词： | 免疫亲和柱 莱克多巴胺 β-受体激动剂 检测 |
Preparation and Application of Immunoaffinity Column for Purification of Ractopamine |
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| WANG Di,YANG Shu-ming,LIU Xiao-wei,YU Hong-xia.Preparation and Application of Immunoaffinity Column for Purification of Ractopamine[J].Journal of Instrumental Analysis,2010,29(8). |
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| Authors: | WANG Di YANG Shu-ming LIU Xiao-wei YU Hong-xia |
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| Institution: | WANG Di1,YANG Shu-ming2,LIU Xiao-wei1,YU Hong-xia2(1.Key Laboratory of Production Environment and Agro-product Safety of Ministry of Agriculture and Tianjin Key Laboratory of Agro-environment and Food Safety,Agro-environmental Protection Institute,Ministry of Agriculture,Tianjin 300191,China,2.Institute of Quality Standard and Test Technology for Agri-product,The Chinese Academy of Agricultural Sciences,Beijing 100081,China) |
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| Abstract: | A polyclonal antibody-based immunoaffinity column(IAC) was developed as a cleanup method for the detection of ractopamine(Rac).Anti-rac specific antibody was gotten with immunized New Zealand rabbit.The IgG was isolated from antiserum with HiTrap Protein AHP column(PrA) and SephadexG25 cartridge.The lmmunosorbents were prepared by coupling anti-Rac IgG with sepharose 4B actived by cyanogens bromide.Rac was cleaned up with the prepared immunoaffinity column and detected by high-performance liquid chromatogra... |
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