Preparation of Monoclonal Antibodies to Trimethoprim and ELISA Kit for Rapid Detection

作     者：韩深 吴小胜 贾芳芳 罗晓琴 万宇平 何方洋 Shen HAN;Xiaosheng WU;Fangfang JIA;Xiaoqin LUO;Yuping WAN;Fangyang HE

作者机构：北京出入境检验检疫局北京100026 北京勤邦生物技术有限公司北京102206 北京市食品安全免疫快速检测工程技术研究中心北京102206 

出 版 物：《Agricultural Science & Technology》 (农业科学与技术（英文版）)

年 卷 期：2016年第17卷第10期

页      面：2267-2270,2372页

学科分类：082803[工学-农业生物环境与能源工程] 08[工学] 0828[工学-农业工程] 0804[工学-仪器科学与技术] 

基　　金：Supported by Beijing Municipal Science and Technology Project(Z151100002115059)~~ 

主　　题：Trimethoprim Monoclonal antibodies Enzyme linked immunosorbent assay kit 

摘      要：[Objective] This study was conducted to find out an approach for determining trimethoprim residues in water. [Method] Trimethoprim antigen was prepared through a series of reactions from trimethoprim hapten which was generated through the reaction between trimethoprim and maleic anhydride. And trimethoprim monoclonal antibodies were prepared by animal immune, and used to prepare ELISA kit to detect trimethoprim residues in water. Finally, the limit of detection （LED） of the ELISA kit was determined. [Result] The standard curve covered a concentration range of 0-80 μg/L. The LeD of trimethoprim in water using the ELISA kit was 2.34 μg/kg; the IC50 （half maximal inhibitory concentration） was 4.8 μg/L; the recovery rate of added trimethoprim standard ranged from 60.5% to 79.7%; within-and among-batches RSD was less than 10%. The trimethoprim monoclonal antibody was specific, as the cross-reactivity rate of trimethoprim antibody and diaveridine was less than 1%. The stability tests revealed that the ELISA kit was stable after being stored at 4 ℃ for 12 months. [Conclusion] The results will provide references for controlling the abuse of trimethoprim.