双酚A单克隆抗体的制备及酶联免疫分析方法的建立  

Preparation of Monoclonal Antibody for the Detection of Bisphenol A by Enzyme-Linked Immunoassay

作　　者：许龙[1] 章英[2] 朱立鑫[1] 范艳[1] 赖肖[2] 孟玮[1] 胡娜[1] 刘仁荣[1] 

机构地区：[1]江西科技师范大学生命科学学院,江西南昌330013 [2]南昌市疾病预防控制中心,江西南昌330038

出　　处：《食品科学》2015年第20期202-206,共5页Food Science

基　　金：国家自然科学基金地区科学基金项目(81360429);江西省高等学校科技落地计划项目(GJJ13573/13574);南昌市科技计划项目(洪财企[2012];37号-23)

摘　　要：采用活泼酯法将双酚A的结构类似物双酚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酚A单克隆抗体,成功获得一株分泌抗双酚A单克隆抗体的细胞株3H1,经鉴定抗体属于Ig G1亚型,轻链为κ,并建立了间接竞争酶联免疫分析法。线性范围为1～50 ng/m L,最低检测限为0.43 ng/m L,半数抑制浓度为6.56 ng/m L。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。4,4-Bis （4-hydroxyphenyl） valeric acid （BVA）, a derivative of bisphenol A （BPA）, was chosen as the hapten to prepare the artificial antigen. An active ester method was used for coupling BPA hapten with keyhole limpet hemocyanin （KLH） and ovalbumin （OVA）, respectively. BALB/c mice were immunized with the immune antigen to produce antibody. After cell fusion with PEG method, the hybridoma cell lines generating anti-BPA monoclonal antibodies were obtained. The monoclonal antibody was identified as IgG 1 subtype with K 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/mL. The limit of detection was 0.43 ng/mL, and the ICs0 value was 6.56 ng/mL. The recovery rates of BPA in water ranged from 82.83% to 101.94% with variation coefficients of 2.94%-12.95%.The proposed immunoassay offers a great potential for sensitive and specific detection of BPA in food safety monitoring.

关 键 词：双酚A 单克隆抗体 酶联免疫分析法 

分 类 号：TQ323.5[化学工程—合成树脂塑料工业]