双酚A单克隆抗体的制备及酶联免疫分析方法的建立

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IgG1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/mL，最低检测限为0.43 ng/mL，半数抑制浓度为6.56 ng/mL。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。

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

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 PE G method, the hybridoma cell lines generating anti-BPA monoclonal antibodies were obtained. The
monoclonal antibody was identified as IgG1 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/mL.
The limit of detection was 0.43 ng/mL, and the IC50 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.