Bioassays Capable of Detecting Single Molecules

描述（由申请人提供）：海贝科技公司开发了一种可以实时光学检测单分子事件的检测方法。该分析利用了等离子共振粒子（PRP）的独特特性，这是一种非常明亮，非漂白的亚微米报告标签。测量单个PRPs从固体底物中通过敏感连接分子的切割释放，被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释放格式的灵敏检测(1)检测蛋白酶活性的多肽连接体，以及(2)检测限制性内切酶介导的切割的核酸连接体。此外，我们已经证明，可以检测到与阻断切割位点的连接子结合的蛋白质，或直接与酶结合，抑制酶活性的蛋白质。

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.