伏马菌素B_1单克隆抗体的制备及鉴定

Generation and Identification of the Monoclonal Antibody Against Fumonisin B_1

【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F3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1.60×10-8 mol.L-1;IC50值可达3.589ng.mL-1;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FB1单克隆抗体具有较好亲和力和高特异性,具有良好的应用价值。

【Objective】 Mycotoxins can lead to serious food security issues.This study focused on the generation of the specific monoclonal antibody（McAb） against fumonisin B1.【Method】 The artificial antigen of fumonisin B1 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 F3 was screened.The antibody secreted by F3 was classified into IgG1 and the light chain type was κ;the results of 10% SDS-PAGE showed that the molecular weight of high chain was 50 kD and the light chain was 25 kD,respectively.The titer of McAb was 1：3 200 in supernatant of F3 and 1：51 200 in ascites with indirect enzyme-linked immunosorbent assay.The affinity was 1.60×10-8 mol？L-1 and the value of IC50 was 3.589 ng？mL-1.There were no cross reactions to other mycotoxins.The high specificity of antibody was identified by western blotting.【Conclusion】The McAb against FB1 in this experiment are of sensitivity and specificity.