伏马菌素B1单克隆抗体的制备及鉴定.pdfVIP

中国农业科学 2011,44(20):4302-4308 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2011.20.019 伏马菌素B 单克隆抗体的制备及鉴定 1 王 莹，郑 浩，何成华，张爱华，冯璐璐，皇超英，张海彬 （南京农业大学动物医学院，南京210095 ） 摘要:【目的】真菌毒素可导致严重的食品安全问题。本试验旨在制备可用于检测伏马菌素B1 的特异性单克隆 抗体。【方法】制备伏马菌素B1人工抗原，杂交瘤细胞法筛选杂交瘤细胞株制备腹水型单克隆抗体，并对其特性进 行鉴定。【结果】筛选得到杂交瘤细胞株F3 ，分泌的单克隆抗体亚类为IgG1，轻链类型为κ链；10%变性聚丙烯酰 胺凝胶电泳显示单抗蛋白重链分子质量约为50 kD，轻链分子质量约为25 kD；间接酶联免疫吸附法测定杂交瘤细 胞F -8 -1 细胞培养上清和腹水效价分别为 1:3 200 和 1:51 200；亲和力常数为 1.60×10 mol·L ；IC 值可达 3.589 3 50 -1 ng·mL ；对其它真菌毒素不存在交叉反应；免疫转印法鉴定抗体具有高特异性。【结论】本试验制备FB 单克隆抗 1 体具有较好亲和力和高特异性，具有良好的应用价值。 关键词：伏马菌素B1 ；人工抗原；单克隆抗体；酶联免疫吸附法 Generation and Identification of the Monoclonal Antibody Against Fumonisin B1 WANG Ying, ZHENG Hao, HE Cheng-hua, ZHANG Ai-hua, FENG Lu-lu, HUANG Chao-ying, ZHANG Hai-bin (College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P. R. China) Abstract: 【Objective】Mycotoxins can lead to serious food security issues. This study focused on the generation of the specific monoclonal antibody (McAb) against fumonisin B 1 . 【Method 】 The artificial antigen of fumonisin B 1 was prepared and a hybridoma cell line was screened with method of hybridoma. Monoclonal antibody was generated by murine ascites and identified. 【Result 】 A hybridoma cell line of F 3 was screened. The antibody secreted by F 3 was classified into IgG1 and the light chain type