Department of Entomology, University of California, Davis, California 95616, USA.
Environ Sci Technol. 2009 Oct 15;43(20):7784-90. doi: 10.1021/es9009037.
We developed a selective competitive enzyme-linked immunosorbent assay (ELISA) to monitor environmental and human exposure to polybrominated diphenyl ether BDE-47 that is used as a flame retardant 2,2',4,4'-Tetrabromodiphenyl ether (BDE-47), a dominant PBDE congener of toxicological concern, was the target analyte. To achieve effective hapten presentation on the carrier protein for antibody production, immunizing haptens with a rigid double-bonded hydrocarbon linker introduced at different positions on the target molecule were synthesized as well as coating haptens that mimic a characteristic fragment of the molecule. Rabbit antisera produced against each immunizing antigen were screened against competitive hapten coating antigens. Underoptimized competitive indirect ELISA conditions, the linear detection range in the assay buffer that includes 50% dimethyl sulfoxide was 0.35-8.50 microg/L with an IC50 value of 1.75 microg/L for BDE-47. Little or no crossreactivity (<6%) was observed to related PBDE congeners containing the BDE-47 moiety and other halogenated compounds. Using a magnetic particle-based competitive direct ELISA increased the sensitivity by 10-fold over the indirect ELISA. The ELISA provided quantitative results when performed on small volume/weight samples such as dust furniture foam, and blood/ serum following sample preparation, suggesting a convenient screening tool.
我们开发了一种选择性竞争酶联免疫吸附测定(ELISA),以监测环境和人体对多溴二苯醚 BDE-47 的暴露情况。BDE-47 被用作阻燃剂,2,2',4,4'-四溴二苯醚(BDE-47)是一种具有毒理学关注的主要多溴联苯醚同系物,是目标分析物。为了在载体蛋白上实现有效的半抗原呈现,用于抗体生产的免疫原是在目标分子的不同位置上引入刚性双键烃键合的半抗原,以及模拟分子特征片段的包被半抗原。针对每种免疫原合成的兔抗血清在竞争性半抗原包被抗原上进行筛选。在包括 50%二甲亚砜的检测缓冲液中,优化后的竞争性间接 ELISA 条件下,该测定的线性检测范围为 0.35-8.50 μg/L,BDE-47 的 IC50 值为 1.75 μg/L。对含有 BDE-47 部分和其他卤代化合物的相关多溴联苯醚同系物的交叉反应性很小或几乎没有(<6%)。使用基于磁性颗粒的竞争直接 ELISA 可使间接 ELISA 的灵敏度提高 10 倍。该 ELISA 在经过样品制备后,从小体积/重量的样品(如灰尘、家具泡沫和血液/血清)上进行时可提供定量结果,表明其是一种方便的筛选工具。