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当前位置: 首页 > 文章 > 双酚A单克隆抗体的制备及酶联免疫分析方法的建立 食品科学 2015,36 (20) 202-206
Position: Home > Articles > Preparation of Monoclonal Antibody for the Detection of Bisphenol A by Enzyme-Linked Immunoassay FOOD SCIENCE 2015,36 (20) 202-206

双酚A单克隆抗体的制备及酶联免疫分析方法的建立

作  者：
许龙;章英;朱立鑫;范艳;赖肖;孟玮;胡娜;刘仁荣
单  位：
南昌市疾病预防控制中心;江西科技师范大学生命科学学院
关键词：
双酚A;单克隆抗体;酶联免疫分析法
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。
译  名：
Preparation of Monoclonal Antibody for the Detection of Bisphenol A by Enzyme-Linked Immunoassay
作  者：
XU Long;ZHANG Ying;ZHU Lixin;FAN Yan;LAI Xiao;M ENG Wei;HU Na;LIU Renrong;College of Life Science, Jiangxi Science and Technology Normal University;Nanchang Center for Disease Control and Prevention;
关键词：
bisphenol A;;monoclonal antibody;;enzyme-linked immunoassay
摘  要：
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 antibo dy. After cell fusion with PEG method, the hybridoma cell lines generating anti-BPA monoclonal antibodies were obtained. The monoclonal antibody was identifi ed as Ig G1 subtype with κ 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/m L. The limit of detection was 0.43 ng/m L, and the IC50 value was 6.56 ng/m L. The recovery rates of BPA in water ranged from 82.83% to 101.94% with variation coefficients of 2.94%–12.95%.The propose d immunoassay offer s a great potential for sensitive and specific detection of BPA in food safety monitoring.

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