莱克多巴胺人工抗原及抗体的制备  

Preparation of Ractopamine Artificial Antigen and Antibody

作　　者：孙海新 凌红丽 曹照真[2] 刘围[2] 高亚东 郗日沫[2] 

机构地区：[1]青岛康地恩实业有限公司,山东青岛266111 [2]山东大学化学与化工学院,山东济南250100

出　　处：《食品科学》2009年第9期25-28,共4页Food Science

基　　金：青岛市科技计划项目(06-3-2-4-nsh);国家自然科学基金项目(20675048);国家“863”计划项目(07AA10Z435;2007AA06A407)

摘　　要：莱克多巴胺是一种禁用β-兴奋剂,建立莱克多巴胺药残的快速检测方法是实现对其进行有力监控的有效途径,而莱克多巴胺抗体是快速免疫检测法的基本试剂。本实验用全新的方法研究了莱克多巴胺免疫原的合成,采用对氨基苯甲酸(ABA)和1,4-丁二醇缩水甘油醚(BDE)将莱克多巴胺分别和cBSA、cOVA偶联,合成了莱克多巴胺的免疫原和包被抗原,并对其进行了紫外分析。用免疫原免疫新西兰大白兔,获得了高灵敏度和特异性的莱克多巴胺多克隆抗体,采用间接ELISA法检测抗体的IC50值为4.34ng/ml,所得多克隆抗体效价达到102400。As one of β-agonists, ractopamine has been prohibited in food production and animal husbandry. To enforce regulation, it is necessary to develop a rapid detection system to measure the residue of ractopamine in animal food production. In this study, a new ractopamine immunogen was synthesized by using 4-aminobenzoic acid as a linker to connect hapten with carrier protein, bovine serum albumin （BSA）. Besides the immunogen, a new ractopamine coating antigen was synthesized also by using 1,4-butanediol diglycidyl ether （BDE） as a linker to connect ovalbumin （OVA）. The BSA conjugate was used to immunize New Zealand rabbits and polyclonal antibodies against ractopamine were obtained. The titers of antibodies were up to 102400 by indirect ELISA and the antibodies showed high sensitivity in the heterologous assay using RAC-BDE-cOVA as coating antigen, with an IC50 value of 4.34 ng/ml. The artificial antigen has perfect immunogencity and the obtained antibody has excellent sensitivity.

关 键 词：莱克多巴胺 ELISA 偶联 免疫原 

分 类 号：S859.84[农业科学—临床兽医学]