LUMINESCENCE ASSAY FOR PHARMACEUTICAL SCREENING

用于药物筛选的发光测定

• 批准号: 2449921
• 负责人: A EIDSATH
• 金额: --
• 依托单位: NATIONAL CENTER FOR RESEARCH RESOURCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/2449921
• 关键词: Xenopus oocyte; biomedical automation; biomedical equipment development; calcium indicator; calcium metabolism; luminescence; pharmacokinetics; photomultiplier

The gene for the photoprotein aequorin has been incorporated into several mammalian cell lines. Because the luminescence of this protein reflects the local Ca++ concentration, it is very useful for assaying cellular Ca++ signals that are activated or blocked by drugs of potential therapeutic value. A major experimental difficulty lies in the very low level of light that is emitted, since only one photon is liberated by each molecule of aequorin, and we have calculated that there are only 20,000 molecules per cell. We plan to modify a photon-counting instrument that we built to measure luminescence from frog oocytes. The photon count will be used as an assay to determine the activity of drugs that influence cell Ca++ metabolism. Large numbers of chemicals will be screened for activity, so the instrument will be automated as much as possible. We have the difficult job of injecting small quantities (20 to 50 microliters) of fluids and cell suspensions.

光蛋白aequorin的基因已经被整合到几个 哺乳动物细胞系。因为这种蛋白质的发光反映了 局部的钙离子浓度，对于细胞的分析是非常有用的 潜在药物激活或阻断的Ca++信号 治疗价值。一个主要的实验困难在于非常低的 发射的光的级别，因为只有一个光子被 每个Aequorin分子，我们计算出只有 每个细胞20000个分子。我们计划修改一个光子计数装置 我们建造了一台仪器来测量青蛙卵母细胞的发光。这个 将使用光子计数作为一种测定药物活性的方法 影响细胞内钙代谢的物质。大量的化学品将被 对活动进行筛选，因此仪器的自动化程度将与 有可能。我们有一项困难的工作，那就是注射少量(20 至50微升)液体和细胞悬浮液。