双酚A单克隆抗体的制备及酶联免疫分析方法的建立.pdfVIP

202 2015, Vo1.36, No .20 自自融事 ※安全检测 双盼A单克隆抗体的制备及酶联兔疫 分析方法的建主 许龙1 ，章英2 ，朱立鑫1 ，范艳1 ，赖肖2 ，孟讳1 ，胡娜1 ，刘仁荣M (1.江西科技师范大学生命科学学院，江西南昌 3300曰; 2.南昌市疾病预防控制中心，江西南昌 330038) 摘 要z 采用活泼酶法将双盼A的结构类似物双酸酸与载体蛋白偶联制备人工抗原，用制备的人工抗原免疫BALB/c 小鼠，采用聚乙二醇法进行细胞融合制备双盼A单克隆抗体，成功获得一株分泌抗双盼A单克隆抗体的细胞株 3Hl ，经鉴定抗体属于IgGl亚型，轻链为1(，并建立了间接竞争酶联免疫分析法。线性范围为1 ~50 ng/r毡，最低检 测限为0.43 nglmL，半数抑制浓度为6.56 nglmL。回收率为82.83%~101.94%，变异系数为2.94%~ 12.95%。该方法 具有较高的灵敏度和特异性，具有良好的应用前景。 关键词:双盼A; 单克隆抗体:酶联免疫分析法 Preparation of Monoclonal Antibody for the Detection of Bisphenol A by Enzyme-Linked Immunoassay 2 2 XU Long , ZHANG Ying , ZHU Lixin , FAN Yan , LAI Xiao , MENG Wei , HU Na , LIU Renrong* (1. College ofLife Science, Jiangxi Science and Technology Norma1 University , Nanchang 330013 , China; 2. Nanchang Center for Disease Control and Prevention , Nanchang 330038 , China) Abstrad: 4 ,4-Bis (4-hydroxyphenyl) va1eric acid (BVA) , a derivative of bisphenol A (BPA) , was chosen as 由e hapten to prepare the artificia1 antigen. An active ester method was used for coupling BPA hapten with keyhole limpet hemocyanin (KLH) and ova1bumin (OVA) , res严ctively. BALB/c mice were immunized with the immune antigen to produce antibody. After cell 缸sion with PEG method, the hybridoma ce111ines generating anti-BPA monoclonal antibodies were obtained. The monoclonal antibody was identified as IgG 1 subtype with 1( light chain. An indirect competitive enzyme-linked immunoassay (ELISA) was set up using the monoclonal antibody. The linear range of the inhibition curve was between 1 and 50 ng旭L The limit of detection was 0.43 nglmL, and the ICso va1ue was 6.56 nglmL. The recovery rates of BPA in water ranged from 82.83% to 1