CCD-BASED FLUORESCENCE POLARIZATION ASSAY PLATE READER

基于 CCD 的荧光偏振分析读板机

• 批准号: 6143138
• 负责人: CLIFFORD C HOYT
• 金额: $ 41.51万
• 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/6143138
• 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.

制药公司目前正在筛选数以百万计的化合物，寻找有希望的候选药物，比如与细胞受体结合的高亲和力药物。应用于这一挑战的许多结合测定法（通常是基于荧光的）都存在限制其使用的缺点。一种检测荧光偏振（FP）变化的技术比其他检测方法有几个优势，包括改进配体和受体之间结合亲和力的量化，由于其同质（混合-读取）格式，增加了简单性和降低了成本。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，操作相对缓慢。CRI开发的“对称FP”方法将大大提高分析仪器的灵敏度和速度，并使基于FP的分析方法成为高通量筛选方法的前沿。我们建议继续开发FP高密度板阅读器。与现有方法相比，第一阶段和随后的工作已经证明，使用标准塑料微滴板和低至1 μ 1的样品体积，灵敏度提高了100倍，板读取速度提高了4倍以上。该仪器对于开发针对体内低水平表达的临床重要细胞受体的药物特别有用。