沙丁胺醇人工抗原的合成及抗体制备  

Preparation of Salbutamol Artificial Antigen and Antibody

作　　者：孙海新 凌红丽 张玉兰[2] 曹照真[2] 高亚东 郗日沫[2] 

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

出　　处：《中国生物工程杂志》2009年第2期48-53,共6页China Biotechnology

基　　金：国家“863”计划(07AA10Z435,2007AA06A407);国家自然科学基金(20675048);青岛市科技计划项目-食品安全专项(06-3-2-4-nsh)资助项目

摘　　要：沙丁胺醇是一种β-兴奋剂,常被很多畜禽水产养殖户非法用于动物养殖。为建立沙丁胺醇在食品中残留的快速检测方法,研究了沙丁胺醇免疫原的合成和抗体的制备方法。采用对氨基苯甲酸法合成了沙丁胺醇(SAL)免疫原SAL-cBSA,采用重氮化法合成的克伦特罗(CL)偶合物CL-cOVA作为包被抗原,用紫外光谱法分析了所合成免疫原和包被抗原。用免疫原SAL-cBSA免疫新西兰大白兔获得多克隆抗体,抗体效价达到32000。采用间接ELISA法检测抗体IC50值为8.79ng/ml,SAL的浓度在1ng/ml～100ng/ml区间时,SAL与对抗体的竞争结合力呈直线关系。表明所制备的沙丁胺醇免疫原具有良好的免疫原性,所制备的抗体拥有很高的灵敏度。To develo Pa rapid analytical method to detect residue of salbutamol in food, the antigen and antibody of salbutamol were prepared. A new salbutamol immunogen was synthesized by using 4-aminobenzoic acid as a linker to connect hapten with carrier protein bovine serum albumin （cBSA）, whereas the coating antigen was synthesized using elenbuterol conjugated with cOVA. UV spectrum analysis proved that the hapten salbutamol was conjugated successfully to cBSA. The cBSA conjugate was used to immunize New Zealand rabbits and polyclonal antibodies against salbutamol were obtained. Titers of antibodies were determined to be 32000 by indirect ELISA and the antibodies showed high sensitivity in the heterologous assay using clenbuterol-cOVA as a coating antigen,with an IC50 value of 8.97ng/ml toward salbutamol. As the concentration of salbutamol increasing from lng/ml to 100ng/ml, the combined degree of sabutamol to antibodies is obviously linear with the concentration of sabutamol. These results showed that the artificially synthesized antigen has perfect immunogeneity and the antibody obtained has high quality.

关 键 词：沙丁胺醇 ELISA 偶联 抗原 抗体 

分 类 号：S859.84[农业科学—临床兽医学]