免疫偶联物设计、合成及间接ELISA方法快速检测纯净水中双酚A的研究
Design and synthesis of immunoconjugates and development of an indirect ELISA for rapid detection of bisphenol A in purified water
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摘要: 设计并合成了双酚A（BPA）的免疫抗原和包被抗原,制备抗BPA多克隆抗体,并建立了间接ELISA方法用于检测瓶装纯净水中BPA的残留。首先,合成了免疫抗原DPA-BSA,并免疫新西兰大白兔得到抗BPA多克隆抗体。其次,合成结构与DPA-OVA有较大差别的包被抗原HPPA-OVA作为"次级结构包被抗原"以提高ELISA方法的灵敏度。基于"次级结构包被原"的策略,建立了检测BPA的间接ELISA方法。该方法的IC50值为1.88ng/mL,最低检测限为0.08ng/mL。最后,用于瓶装纯净水样品中BPA的检测,其平均回收率为89.6%,平均变异系数为8.8%。因此,该方法可用于水样中BPA的快速检测。Abstract: The immunoconjugates were designed, synthesized and then used to develop a rapid, specific and sensitive indirect ELISA method to detect residues of bisphenol A ( BPA) in bottled purified water. The immunogen DPA-BSA was first synthesized and used to immunized to rabbits to produce a polyclonal antibody against BPA. Furthermore, HPPA-OVA structurally different from DPA-OVA was designed and used as a “substructural coating antigen” to improve the sensitivity of an indirect ELISA analysis for BPA detection. Based on the “substructural coating antigen” concept, an optimized indirect ELISA method was developed that exhibited good specificity and high sensitivity for detecting BPA, with IC50 of 1.88ng/mL and detection limit of 0.08ng/mL. Finally, simple and efficient analysis of BPA samples in bottled purified water showed that the average recovery rate of the ELISA analysis was 89.6%, with the average coefficient of variation 8.8%. Thus, the developed indirect ELISA method exhibited the potential for a rapid detection of BPA residues in water.
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