Multiresidue Method for Analysis of beta Agonists in Swine Urine by Enzyme Linked Receptor Assay Based on beta(2) Adrenergic Receptor Expressed in HEK293 Cells

文献类型： 外文期刊

第一作者： Wang, Jian

作者： Wang, Jian;She, Yongxin;Wang, Miao;Jin, Maojun;Li, Yongfei;Wang, Jing;Wang, Jian;Liu, Yuan

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2015 年 10 卷 9 期

页码：

收录情况： SCI

摘要： A novel enzyme-linked receptor assay (ELRA) based on beta(2)-adrenergic receptor (beta(2)-AR) has been developed for rapid and high-throughput detection of beta-adrenergic agonists (beta-agonists) in urine. Human embryonic kidney cells (HEK293) were introduced as the expression system to enhance the functionality of the recombinant beta(2)-AR, and the attempt to detect beta-agonists in swine urine using such approaches was accomplished unprecedentedly. In this article, a recombinant porcine beta(2)-AR was produced in the inner membrane of HEK293 cells and purified from crude membrane protein by nickel-nitrilotriacetic acid affinity chromatography. After activity identification, the recombinant receptor was used in the development of direct competitive ELRA. Several parameters such as blocking buffer and blocking process were optimized and the performance of the system was determined. The IC50 concentrations of clenbuterol, salbutamol, and ractopamine were 34, 53 and 63 mu g/L, and the average recovery rates were 68.2%, 60.3% and 65.5%, respectively. ELRA based on beta(2)-AR shows a series of advantages such as safety, easy operation, and high efficiency, making it promising for the rapid screening of beta-agonists in animal urine.

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