Detection of xenoestrogens in serum after immunoprecipitation of endogenous steroidal estrogens.
Detection of xenoestrogens in serum after immunoprecipitation of endogenous steroidal estrogens.
复制标题
DOI：
10.1289/ehp.02110791
复制
发表时间：
2002-08
影响因子：
10.4
通讯作者：
Lasley BL
中科院分区：
文献类型：
--
作者：
Natarajan K;Overstreet JW;Rogers JM;Denison MS;Chen J;Lohstroh PN;McConnell DS;Lasley BL
在这篇文章中，我们报告了一个简单而有效的方法检测生物样品中的非甾体雌激素。该方法使用雌二醇（E2）的多克隆抗体来免疫沉淀这些主要的生物活性甾体雌激素，留下非甾体雌激素，然后在基于细胞的雌激素受体激动剂的转录激活生物测定中检测。免疫沉淀法有效地去除99%的放射性标记的E2和雌酮（E1）从人血清。在实验中，超生理浓度的E2和E1的人血清中，所有的免疫反应性雌激素仍然被删除的免疫沉淀协议。我们进行了这种方法的体内验证研究，其中，我们治疗雌性猕猴与异雌激素壬基酚（NP），在卵泡晚期的月经周期。我们使用治疗前后采集的血液样本来评估和表征内源性和外源性血清雌激素。E2的免疫测定未检测到治疗猴中的NP。基于细胞的生物测定法也没有检测到NP的雌激素活性，因为其饱和的内源性血清甾体雌激素。然而，当通过免疫沉淀去除甾体雌激素时，我们在生物测定中检测到NP的雌激素活性。因此，这种方法适用于检测血清样品中的外源性非甾体雌激素。
In this article we report a simple and efficient method for detecting nonsteroidal estrogens in a biologic sample. This method uses polyclonal antibodies to estradiol (E2) to immunoprecipitate these major biologically active steroidal estrogens, leaving behind the nonsteroidal estrogens, which are then detected in a cell-based transcriptional activation bioassay for estrogen receptor agonist. The immunoprecipitation method efficiently removed 99% of radiolabeled E2 and estrone (E1) from human serum. In experiments in which supraphysiologic concentrations of E2 and E1 to human serum, all of the immunoreactive estrogens were still removed by the immunoprecipitation protocol. We carried out an in vivo validation study of this method in which we treated female macaques with the xenoestrogen nonylphenol (NP), during the late follicular phase of the menstrual cycle. We used blood samples collected before and after treatment to evaluate and characterize endogenous and exogenous serum estrogens. An immunoassay for E2 did not detect the NP in treated monkeys. The cell-based bioassay also did not detect the estrogenic activity of NP because of its saturation by the endogenous serum steroidal estrogens. However, when steroidal estrogens were removed by immunoprecipitation, we detected the estrogenic activity of NP in the bioassay. Thus, this approach is appropriate for detecting exogenous, nonsteroidal estrogens in serum samples.