沙丁胺醇多克隆抗体的制备与鉴定_张赛.pdf

现代食品科技 Modern Food Science and Technology 2013, Vol.29, No.3 沙丁胺醇多克隆抗体的制备与鉴定 张赛 1，许青华 1，李文美 1,2，何小维 1 （1.华南理工大学轻工与食品学院，广东广州 510640） （2.广州万孚生物技术有限公司国家地方联合工程实验室，广东广州 510530） 摘要：为了对沙丁胺醇进行酶联免疫吸附法（ELISA）测定，通过四种方法制备沙丁胺醇衍生物，分别将其与牛血清蛋白（BSA）、 卵清蛋白（OVA）偶联，制备人工完全抗原，采用紫外分光光度法测定蛋白质浓度并计算偶联比，用所制备的四种免疫原免疫新西 兰白兔获得抗沙丁胺醇（SAL）抗体，每一种抗体都用四种包被抗原进行配对筛选，据此得???最佳的抗体-包被抗原的组合，对筛选 出的抗体用正辛酸-硫酸铵法进行纯化，采用间接 ELISA 法测定多克隆抗体（pAb）的效价并进行特异性鉴定。结果表明：成功偶联 了完全抗原，四种免疫原的偶联比分别为 9:1、6:1、6:1、12:1，得到最佳的抗体-包被抗原的组合，筛选出的抗体效价达到 1:64000， 此抗体对克伦特罗和特布他林的交叉反应率分别为 144%和 169%，对其他类似物几乎没有交叉反应。本研究为沙丁胺醇、克伦特罗 和特布他林多残留酶联免疫检测法的建立奠定了基础。 关键词：沙丁胺醇；人工抗原；多克隆抗体 文章篇号：1673-9078(2013)3-563-567 Preparation and Identification of Polyclonal Antibodies against Salbutamol ZHANG Sai1, XU Qing-hua1, LI Wen-mei1,2, HE Xiao-wei1 (1.College of Light Industry and Food Science, South China University of Technology, Guangzhou 510640, China) (2.National Local United Engineering Laboratory, Wondfo Biotech Co., Ltd, Guangzhou 510530, China) Abstract: In order to prepare the polyclonal antibody against Salbutamol(SAL), SAL derivatives were prepared by four kinds of reactions, then they were coupled to BSA and OVA, synthesizing complete antigen. The protein concentration was determined by Ultraviolet spectrophotography and the coupling ratios of four immunogens were calculated. New Zealand white rabbits were immunized by four immunogens to develop anti-SAL antibodies. All the antibodies and coating antigenes were screened and then selected the best combination. The polyclonal antiserum was purified b