BIOSENSOR STUDIES OF ESTROGENIC COMPOUNDS WITH HER-A & B

1999年，环保局将开始对内分泌干扰潜力的化学品进行第一级筛选。筛查包括评估与雌激素受体(HER)的相互作用。有益的植物雌激素和有害的合成雌激素一样束缚着她。需要一种将两者区分开来的方法。IA公司建议将其光纤传感器广泛而独特地应用于检测雌激素模拟物，以测试解决这一问题的创新方法的可行性。这项拟议的工作将检验这样一种假设，即这种区别与HER-α和HER-β与雌激素反应元件结合和分离的动力学上的可测量差异有关。根据这项提议的具体目标，詹姆斯·威特利夫博士将建立、标记和表征HER-β的荧光团供应。IA Inc.将制造具有雌激素配体或雌激素反应元件的纤维传感器。利用光纤传感器研究植物雌激素、外源雌激素和内源性雌激素对雌激素反应元件与HER-α和β结合动力学的影响。将研究10-10-10-5M之间的浓度，并与许多雌激素类化合物高剂量和低剂量生物效应差异的文献报道相关。拟议的商业应用化学工业和环境保护局的检测实验室需要一种成本相对较低(10,000美元)的便携式仪器和试剂，用于测试环境中的化学物质是否具有雌激素活性。这项研究将提供界定该仪器适用范围的数据。

In 1999 the EPA will begin Tier 1 screening of chemicals for endocrine disrupting potential. Screening includes assessment of interactions with estrogen receptor (hER). Beneficial phytoestrogens bind hER as do harmful synthetic estrogens. A method is needed which distinguishes one from the other. IA, Inc. proposes an extensive and unique application of its fiber optic sensor for detecting estrogen mimics so as to test the feasibility of an innovative approach to this problem. The proposed work will test the hypothesis that this distinction is related to measurable differences in the kinetics with which hER- alpha and hER-beta bind to and dissociate from the estrogen response element. Under the specific aims of this proposal Dr. James Wittliff will establish, fluorophore-label and characterize a supply of hER-beta. IA Inc. will create fiber sensors possessing either estrogenic ligand or estrogen response element. The effects of phytoestrogens, xenoestrogens, and endogenous estrogens on the kinetics of binding between estrogen response element and hER-alpha and beta will be studied using the fiber optic sensor. Concentrations between 10-10-10-5M will be studied and related to literature reports of differences in the biological effects of high and low doses of many estrogenic compounds. PROPOSED COMMERCIAL APPLICATION A relatively low cost (10,000 dollars) portable instrument with reagents for testing chemicals in the environment for estrogenic activity is needed by the chemical industry and by EPA testing laboratories. This research will provide data which defines the scope of application of the instrument.