抗呋喃唑酮单克隆抗体的制备及其应用.pdf

※生物工程 食品科学 2016, Vol.37, No.03 157 抗呋喃唑酮单克隆抗体的制备及其应用 1 1 1,2,* 3 3 陈荫楠 ，陈 华 ，石贤爱 ，叶小军 ，樊海平 （1.福州大学生物科学与工程学院，福建 福州 350108 ；2.福建省医疗器械和医药技术重点实验室，福建 福州 350002 ； 3.福建省淡水水产研究所，福建 福州 350002 ） 摘  要：为实现呋喃唑酮（furazolidone ，FZD ）的快速定量检测，本研究制备了抗FZD全抗原的单克隆抗体，建立 了抗FZD 的单克隆抗体酶联免疫检测方法。结果表明，FZD单克隆抗体在质量浓度为10～500 ng/mL范围具有较好 的线性，IC50值为0.06 μg/mL ，最低检测限为6.92 ng/mL ，对于其他硝基呋喃类抗生素及其代谢物均不存在交叉反 应。该方法重现性较好，平均误差为6.54%，回收率为76.84%～88.31%。 关键词：呋喃唑酮；单克隆抗体；酶联免疫检测 Preparation and Application of Monoclonal Antibody against Furazolidone 1 1 1,2,* 3 3 CHEN Yinnan , CHEN Hua , SHI Xianai , YE Xiaojun , FAN Haiping (1. College of Biological Science and Engineering, Fuzhou University, Fuzhou 350108, China; 2. Fujian Key Laboratory of Medical Instrument and Pharmaceutical Technology, Fuzhou 350002, China; 3. Fresh Water Fisheries Research Institute of Fujian Province, Fuzhou 350002, China) Abstract: Based on the hapten of furazolidone (FZD), a monoclonal (mAb)-based enzyme-linked immunosorbent assay (ELISA) method for the detection of FZD was established after the preparation of mAb against FZD-bovine serum albumin (BSA). The linear range for furazolidone detection was 10–500 ng/mL, and the IC50 was 0.06 μg/mL with a lowest detection limit of 6.92 ng/mL. Meanwhile, the mAb showed almost no cross reactivity with other nitrofurans or their metabolites. This method had good reproducibility, with an average error of 6.54%. In tested samples, the recovery for furazolidone additio