Instituto Interuniversitario de Investigación en Biongeniería y Tecnología Orientada al Ser Humano, Universidad Politécnica de Valencia, Spain.
J Environ Sci Health B. 2011;46(6):509-17. doi: 10.1080/03601234.2011.583871.
The aim of this work was the development of monoclonal antibodies (MAbs) and highly sensitive immunoassays (ELISAs) to bisphenol A (BPA), a well-known endocrine disruptor able to migrate from the internal coating of cans to food contained inside, particularly vegetables. To produce MAbs to BPA, four synthetic compounds were conjugated to proteins and used as immunizing haptens in mice. By applying hybridoma technology, several MAbs were produced and selected. These antibodies were characterized in the conjugate-coated and in the antibody-coated formats, using both homologous and heterologous conjugates. Three indirect ELISA based on the MAbs showing the highest affinity to BPA were selected. The limit of detection of the most sensitive ELISA was 0.22 nM (0.05 ng/mL), with an I₅₀ value of around 1 nM (0.23 ng/mL). An homologous ELISA based on the MAb BPAB-11 was applied to the simple, direct determination of BPA in the liquid portion of canned artichoke, peas, and sweet corn. Only sample dilution in an appropriate saline buffer was required to minimize matrix effects and to enter the ELISA working range. Recovery and precision of the method were evaluated by spiking the liquid portion of these cans with BPA at 20, 50, and 100 ng/mL. Coefficients of variation were below 20% in most cases. With regard to recovery, the analytical data obtained were also acceptable. This immunoassay has therefore proved its potential as a new tool for the rapid, sensitive and accurate determination of BPA in canned food.
本工作旨在开发针对双酚 A（BPA）的单克隆抗体（MAb）和高灵敏度免疫分析（ELISA），BPA 是一种众所周知的内分泌干扰物，能够从罐头的内部涂层迁移到内部的食物中，尤其是蔬菜。为了生产针对 BPA 的 MAb，将四种合成化合物与蛋白质缀合，并用作小鼠的免疫原。通过应用杂交瘤技术，生产并选择了几种 MAb。使用同源和异源缀合物，在缀合物包被和抗体包被的格式中对这些抗体进行了表征。选择了基于对 BPA 具有最高亲和力的 MAb 的三种间接 ELISA。最敏感的 ELISA 的检测限为 0.22 nM（0.05 ng/mL），I₅₀ 值约为 1 nM（0.23 ng/mL）。基于 MAb BPAB-11 的同源 ELISA 被应用于简单、直接地测定罐装朝鲜蓟、豌豆和甜玉米液体部分中的 BPA。只需将样品稀释在适当的盐缓冲液中，以最小化基质效应并进入 ELISA 的工作范围。通过在这些罐头的液体部分中以 20、50 和 100 ng/mL 的浓度添加 BPA 来评估方法的回收率和精密度。在大多数情况下，变异系数低于 20%。就回收率而言，获得的分析数据也是可以接受的。因此，该免疫测定法已被证明具有作为快速、灵敏和准确测定罐装食品中 BPA 的新工具的潜力。
