QCM Quantification of Endocrine Disruptor Activity

描述(申请人提供)：具有雌激素样活性的环境制剂是一个主要的公共卫生问题。尽管已经开发了各种方法来检测与雌激素受体(ER)的结合，并开发了其他方法来监测雌激素敏感基因转录的后续影响，但一个替代方法验证机构间协调委员会(ICCVAM)最近报告了所有可用的方法都存在困难。 我们正在开发石英晶体微天平(QCM)平台技术，用于复杂大分子相互作用的定量高通量分析。本项目的重点是开发QCM生物传感器，用于快速定量内分泌干扰物对雌激素受体(ER)、辅助激活因子或辅助抑制因子和雌激素反应元件(ERE)之间相互作用的影响。 QCM已被应用于研究蛋白质与小分子的相互作用、蛋白质与蛋白质的相互作用以及DMA杂交。预计QCM将提供迄今为止无法获得的关于内分泌干扰物对ER与ERE的动力学和亲和力的影响的定量信息，以及关于内分泌干扰物对ER与辅助激活因子、辅助抑制因子和其他转录因子之间相互作用的影响的可比信息，这些转录因子的相互作用调节许多细胞类型和生物体中的雌激素反应基因。QCM的主要优点是灵敏度高、成本低、实时无标记、定量分析。鉴于美国国会的授权和世界卫生组织对化学品内分泌干扰物测试的建议，我们预计市场对ERE生物传感器技术的需求强劲。

DESCRIPTION (provided by applicant): Environmental agents with estrogen-like activity are a major public health issue. Although a variety of assays have been developed to detect binding to the estrogen receptor (ER) and other assays have been developed to monitor subsequent effects on estrogen sensitive gene transcription, an Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) recently reported difficulties with all available methods. We are developing quartz crystal microbalance (QCM) platform technology for the quantitative high-throughput analysis of complex macromolecular interactions. The focus of this project is the development of QCM biosensors for rapid quantification of the influence of endocrine disrupters on the interactions among the estrogen receptor (ER), coactivators or corepressors and the estrogen response element (ERE). QCM has been applied to study interactions of proteins with small molecules, protein-protein interactions, and DMA hybridization. It is expected that QCM will provide heretofore inaccessible quantitative information regarding effects of endocrine disrupters on the kinetics and affinity of the ER for the ERE as well as comparable information about the influence of endocrine disrupters on interactions between the ER and coactivators, corepressors, and other transcription factors whose interactions regulate estrogen-responsive genes in many cell types and organisms. Major advantages of QCM include high sensitivity, low cost, real time label-free, quantitative analysis. Given the U.S. congressional mandate and World Health Organization recommendations for endocrine disrupter testing of chemicals, we anticipate strong market demand for ERE biosensor technology.