胶体金免疫层析卡优化及其与ELISA法检测克伦特罗残留的比较  

Optimization of Colloidal Gold Immunochromatographic Card and Comparison with ELISA for the Detection of Clenbuterol Residues

作　　者：刘颖沙 刘昭彤[2] 邢维维 鲁瑶 岳彩洋 杨洋 LIU Yingsha;LIU Zhaotong;XING Weiwei;LU Yao;YUE Caiyang;YANG Yang(College of Bioengineering,Yangling Vocational Technical College,Yangling,Shaanxi 712100,China;Xi'an Branch of China Academy of Space Technology,Xi'an,Shaanxi 710100,China;Beijing Weide Weikang Biotechnology Co.,Ltd.,Beijing 100095,China)

机构地区：[1]杨凌职业技术学院生物工程分院,陕西杨凌712100 [2]中国空间技术研究院西安分院,陕西西安710100 [3]北京维德维康生物技术有限公司,北京100095

出　　处：《热带农业科学》2023年第1期62-67,共6页Chinese Journal of Tropical Agriculture

基　　金：陕西省教育厅2021年度一般专项科研计划项目(No.21JK1001);杨凌职业技术学院2022年院内基金项目研究成果(No.ZK22-47)。

摘　　要：盐酸克伦特罗俗称“瘦肉精”,国家明令禁止使用,市面上最常见的瘦肉精快速检测方法即为快速检测试纸条,因其特异性和灵敏度较低,容易造成结果的不准确。本文优化克伦特罗单抗的制备方法,单克隆抗体的免疫球蛋白亚类为IgG1,分子量为148.5 kDa,染色体数目为83~91条,亲和常数为2.51×10^(8)M^(-1),得到最佳的标记条件pH=8、最佳抗体标记量为10μL/mL的120%(12μL/mL)抗体标记量、最佳离心力为8000 r/min,对样品垫、吸水纸、NC膜进行探索与优化,本研究研制的克伦特罗检测卡检出限为3 ng/mL,与莱克多巴胺、沙丁胺醇和妥布特罗无交叉反应,特异性好,与酶联免疫吸附法测定值符合率良好。Clenbuterol hydrochloride,commonly known as"leptin"is banned by the state.The most common rapid detection method of leptin in the market is the fast detection strip.Because of its low specificity and sensitivity,it is easy to cause inac-curate results.In this paper,the preparation method of clenbuterol monoclonal antibody was optimized,the immunoglobulin subclass of monoclonal antibody was IgG1,the molecular weight was 148.5 kDa,the number of chromosomes was 83-91,the affinity constant was 2.51×10^(8)M^(-1),the optimal labeling conditions were pH=8,the optimal antibody labeling amount was 120%(12μL/mL)of 10μL/mL,and the optimal centrifugal force was 8000 r/min.The detection limit of the clenbuterol de-tection card was 3 ng/mL.There was no cross-reaction with ractopamine,salbutamol,and tobuterol,and the specificity was excellent.The results were in good agreement with those measured by enzyme-linked immunosorbent assay.

关 键 词：盐酸克伦特罗 检测 单克隆抗体 准确度 特异性 酶联免疫吸附法 

分 类 号：TS251.7[轻工技术与工程—农产品加工及贮藏工程]