CCD-BASED FLUORESCENCE POLARIZATION ASSAY PLATE READER

基于 CCD 的荧光偏振分析读板机

• 批准号: 6392362
• 负责人: CLIFFORD C HOYT
• 金额: $ 33.46万
• 依托单位: CAMBRIDGE RESEARCH AND INSTRUMENTATION
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-15 至 2002-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6392362
• 关键词: biomedical equipment development; charge coupled device camera; computer program /software; computer system design /evaluation; drug screening /evaluation; fluorescence polarization; image processing; liquid crystal; method development

Pharmaceutical companies currently screen millions of compounds looking for promising drug candidates such as agents that bind with high affinity to cellular receptors. Many binding assays (often fluorescence-based) being applied to this challenge suffer from drawbacks that limit their use. A technique examining changes in fluorescence polarization (FP) promises several advantages over other assays, including improved quantification of binding affinity between ligand and receptor, and increased simplicity and lower cost, due to its homogeneous (mix-and-read) format. Currently available FP-based instruments are not ideal, however, because of their low sensitivity and relatively slow operation. An approach, "Symmetric FP", developed at CRI, will lead to analytical instruments with greatly improved sensitivity and speed and should bring FP-based assays to the forefront of high-throughput screening methods. We propose to continue development of an FP high-density plate reader. Phase I and subsequent work have demonstrated 100-fold improvement in sensitivity using standard plastic microtitre plates and sample volumes as low as 1 mu1, and a more than 4-fold increase in plate-reading speed, compared to existing methods. The instrument will be particularly useful for developing drugs targeting clinically significant cellular receptors expressed at low levels in vivo. PROPOSED COMMERCIAL APPLICATION: Our plate reader would be used for drug discovery by pharmaceutical companies. The larger companies, of which there are at least 50, have between 20 and 50 drug screening programs ongoing at any given time. We expect that most of their existing ligand-screening instrumentation will be replaced by higher throughput, more cost-effective instruments, notably the proposed CRI FP platform. Conservative estimates suggest a potential market for 6-18 million dollars.

制药公司目前筛选了数百万种化合物，寻找有希望的候选药物，如与细胞受体具有高亲和力的药物。应用于这一挑战的许多结合测定（通常基于荧光）存在限制其使用的缺点。检查荧光偏振（FP）变化的技术承诺优于其他测定的几个优点，包括配体和受体之间的结合亲和力的改进的定量，以及由于其同质（混合和读取）形式而增加的简单性和更低的成本。然而，目前可用的基于FP的仪器并不理想，因为它们的灵敏度低且操作相对较慢。CRI开发的一种方法“对称FP”将使分析仪器的灵敏度和速度大大提高，并将基于FP的测定带到高通量筛选方法的最前沿。我们建议继续开发FP高密度酶标仪。第一阶段和随后的工作已经证明，使用标准塑料微量滴定板和样品体积低至1 μ l的灵敏度提高了100倍，与现有方法相比，读板速度提高了4倍以上。该仪器将特别适用于开发靶向体内低水平表达的临床显著细胞受体的药物。 拟定商业应用： 我们的酶标仪将被制药公司用于药物发现。大公司，其中至少有50个，有20至50个药物筛选计划正在进行中的任何给定的时间。我们预计，他们现有的大多数配体筛选仪器将被更高的通量，更具成本效益的仪器，特别是拟议的CRI FP平台所取代。保守估计，潜在市场为600万至1800万美元。