基于ELISA的受体竞争结合法检测环境雌激素

Competitive Estrogen Receptor Binding Based ELISA for Detection of Estrogen in Environment

为建立环境雌激素的体外快速筛选方法,采用免疫测定介导受体竞争结合试验定量检测环境雌激素.将17β-雌二醇-BSA固定在微孔板上,环境雌激素与微孔板上的17β-雌二醇-BSA竞争性地与雌激素受体结合,与微孔板上17β-雌二醇-BSA结合的受体与雌激素受体的抗体、雌激素受体抗体的二抗形成一个三明治形式的复合物,通过显色剂显色,最后用比色法对试验结果进行定量.结果表明,环境雌激素的浓度与溶液吸光度值成负相关.定量研究可以得到环境雌激素剂量-效应关系曲线,在一定浓度范围内(10ng·L-1～100μg·L-1),溶液的吸光度值与环境雌激素的浓度直接呈现良好的线性关系(R2=0.9583).利用这种方法能检测到的标准品雌二醇的最低浓度为10ng·L-1.以上结果表明,本方法检测环境雌激素操作简便,具有广泛的应用前景.

In order to develop a rapid and sensitive bioassay method for the detection of environmental estrogens, we present a simple method for quantitative assessment of environmental estrogens, based on the competitive estrogen receptor（ER）mediated immunoassay in microplates. Estrogen, ER, anti-ER antibody and secondary antibody labeled horseradish peroxidase（HPR）can form a sandwich complex. The signals of the result were produced by color reagent, and record with colorimetric absorbance. The assay result showed that 10ng·L-1 of 17β-estradiol could be detected and have a linear relationship between the absorbance and the estrogen concentrations in the range from 10ng·L-1 to 100μg·L-1（R2=0.9583）. It indicated that this method provides a simple way to quantification environmental estrogens in samples and could be wildly used.