Bioassays Capable of Detecting Single Molecules

描述(申请人提供)：贝壳科技公司开发了一种可以光学实时检测单分子事件的分析方法。该检测方法利用了等离子体共振颗粒(PRP)的独特性质，这是一种非常明亮的、不漂白的亚微米报告标记。通过裂解敏感的连接物分子从固体底物上测量单个PRPS的释放，可用作酶活性的超灵敏检测器。我们设计并实现了两种基于PRP释放形式的灵敏分析方法：(1)检测蛋白水解酶活性的多肽连接物；(2)检测限制性内切酶介导的切割的核酸连接物。此外，我们已经证明，可以检测到与阻断裂解位点的连接子结合的蛋白质，或者直接与酶结合、抑制酶活性的蛋白质。 开发的分析方法将简单易用，只需要非常少量的样品，并且将可以实时检测。我们期望我们的SBIR计划的结果，与工业合作伙伴合作，使我们能够创造商业产品销售给研究和临床生物医学界。

DESCRIPTION (provided by applicant): Seashell Technology has developed an assay that can optically detect single molecular events in real time. The assay makes use of the unique properties of the plasmon resonant particle (PRP), an extremely bright, non-bleaching, sub-micron reporter label. The measured release of individual PRPs from a solid substrate by cleavage of susceptible linker molecules is used as an ultrasensitive detector of enzymatic activity. We have designed and implemented two sensitive assays based on the PRP release format (1) a polypeptide linker that detects protease activity, and (2) a nucleic acid linker that detects restriction enzyme mediated cleavage. In addition, we have demonstrated that proteins that bind to the linker blocking the cleavage site, or bind directly to the enzyme, inhibiting the enzymatic activity, can be detected. The developed assays will be simple to use, require very small quantities of sample, and will be detectable in real time. We expect the results of our SBIR program, in collaboration with industrial partners, to allow us to create commercial products for sale to both the research and clinical biomedical communities.