NOVEL ASSAY FOR G PROTEIN COUPLED RECEPTORS

该项目的目标是开发一个系统， 用于快速基于细胞的测定的仪器、试剂和方法 配体与G蛋白偶联受体的结合并用于快速评估 的细胞反应。该方法是基于测量荧光 荧光配体缀合物之间的共振能量转移（FRET） （受体）和G蛋白偶联受体固有标记 绿色荧光蛋白（供体）。该方法的一个关键创新是 使用一种新的荧光寿命传感技术，最近 用于测量配体结合。也就是说， 配体的荧光寿命的减少被转导， 绿色荧光蛋白。长期目标是发展 紧凑、坚固、灵敏、低成本的高通量仪器 药物发现中的筛选拟议的工作涉及以下方面的需要： 制药公司在竞争压力下， 成功的新治疗产品上市更快， 有效地庞大的铅化合物库 从天然来源如植物、海洋生物和真菌或从 组合技术已经产生了对更好、更快和 更便宜的高通量筛选技术。一个最 药理学干预的重要药物靶点类别是 G蛋白偶联受体（GPCR）。市场潜力 用于GPCR的药物是巨大的。他们已经占到了 今天制造的所有药物，并占最大的目标之一， 人类基因组计划的结果。据我们所知，这 将是第一次利用荧光寿命传感 用于高通量筛选应用。 拟定商业应用： 先导药物发现、细胞和分子的高通量筛选 生物研究仪器，临床诊断仪器。

The objective of this project is to develop a system consisting of instrumentation, reagents and methods for rapid cell-based assays of ligand binding to G protein-coupled receptors and for rapid assessment of cellular response. The approach is based on measuring fluorescence resonance energy transfer (FRET) between fluorescent ligand conjugates (acceptor) and G protein-coupled receptor intrinsically tagged with green fluorescent protein (donor). A key innovation of the approach is the use of a new fluorescence lifetime sensing technology, recently developed by Ciencia, for measuring ligand binding. That is, the binding of ligand is transduced as a decrease in the fluorescence lifetime of the green fluorescent protein. The long term objective is to develop compact, robust, sensitive, low-cost instrumentation for high throughput screening in drug discovery. The proposed work addresses the needs of pharmaceutical companies which are under competitive pressures to bring successful new therapeutic products to market faster and more efficiently. The enormous libraries of lead chemical compounds assembled from natural sources such as plants, ocean life, and fungi or from combinatorial techniques have created a need for better, faster and cheaper technologies for high throughput screening. One of the most important classes of drug targets for pharmacological intervention is the family of G-protein coupled receptors (GPCR). The market potential for drugs targeted to GPCR is huge. Already, they comprise about 60% of all drugs manufactured today, and account for one of the largest target families resulting from the Human Genome Project. To our knowledge, this would be the first time for fluorescence lifetime sensing is exploited for high throughput screening applications. PROPOSED COMMERCIAL APPLICATION: High throughput screening in lead drug discovery, cell and molecular biology research instrumentation, clinical diagnostic instrumentation.