Bioassays Capable of Detecting Single Molecules

描述（由申请人提供）：Seashell Technology已开发出一种可真实的实时光学检测单分子事件的检测方法。该检测利用了等离子体共振粒子（PRP）的独特性质，这是一种非常明亮、非漂白的亚微米报告标记。通过敏感连接分子的切割从固体基质中测量的单个PRP的释放被用作酶活性的超灵敏检测器。我们设计并实施了两种基于PRP释放形式的灵敏测定法：（1）检测蛋白酶活性的多肽接头，和（2）检测限制性内切酶介导的切割的核酸接头。此外，我们已经证明，可以检测到与阻断切割位点的接头结合或直接与酶结合从而抑制酶活性的蛋白质。 所开发的检测方法将易于使用，需要非常少量的样品，并且将在真实的时间内被检测到。我们希望我们的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.