Purchase of a Biacore T200 Surface Plasmon Resonance Label-free Detection System

描述(由申请者提供)：需要资金购买Biacore T200台式表面等离子体共振(SPR)无标签检测系统。Wistar研究人员发现迫切需要这项技术，以支持NIH资助的研究，以研究蛋白质：蛋白质、蛋白质：核酸和小分子配体之间的结构功能关系：蛋白质相互作用对我们对肿瘤细胞生物学、治疗、耐药性、肿瘤复发和转移的基本理解具有关键影响。Wistar研究所没有具备Biacore T200功能的仪器，在我们的研究社区内直接访问这项技术作为共享资源是有限的。Wistar分子筛选实验室的建立是为了使其科学家能够在高密度微滴定板中筛选小分子化学库，在我们的研究计划中加强化学生物学和转化医学，并促进新的合作研究和学术-商业伙伴关系。目前的实验室基础设施使研究人员能够通过标记的(即放射性、荧光、化学发光)示踪剂/底物识别影响目标活动的分子。尽管来自标记底物的读数对高通量非常敏感和服从，但它们也容易受到干扰，导致假阳性和阴性发现率增加，并且没有证据表明调节器直接与目标靶点结合。自成立以来，该实验室促进了针对10种不同靶标的小分子筛查的开发和执行，包括伽玛疱疹病毒潜伏期、人乳头瘤病毒癌蛋白、p53和人类端粒酶。这些筛选的结果已经为每个项目确定了2-3个确认的“命中”系列，这些项目已经准备好通过药物化学进行优化。然而，现有的技术(即ITC、分析超速离心法的沉降平衡)产量较低，需要大量稀有或敏感材料来评估小分子与靶之间的相互作用。因此，所要求的Biacore T200将在维斯塔尔研究所分子筛查设施中定位和维护。Biacore T200的灵敏度、多功能性和吞吐量将立即影响NIH资助的6个主要用户和一群次要用户定量评估广泛生物分子相互作用的能力，并开发基于片段的筛选，作为蛋白质功能小分子抑制剂的替代品。一个由维斯塔尔研究所的用户和高级工作人员组成的内部咨询委员会将监督该仪器的操作、维护和公平使用。在本申请中，我们介绍了需要获得表面等离子体共振无标记检测灵敏度的研究项目和初步数据。购买Biacore T200将极大地增强研究人员表征生物分子相互作用的能力，并扩大Wistar分子筛查设备的能力和多功能性，以支持旨在减轻人类疾病负担的变革性研究。

DESCRIPTION (provided by applicant): Funds are requested to purchase a Biacore T200 benchtop Surface Plasmon Resonance (SPR) label-free detection system. Wistar investigators have identified an urgent need for this technology in order to support NIH funded research to study structure function relationships between protein: protein, protein: nucleic acid, and small molecule ligand: protein interactions with critical implications to our fundamental understanding of tumor cell biology, treatment, drug resistance, tumor recurrence, and metastasis. The Wistar Institute does not have an instrument with the capabilities of the Biacore T200 and direct access to this technology within our research community is limited as a shared resource. The Wistar Molecular Screening Laboratory was established to enable its scientists to screen small molecule chemical libraries in high-density microtiter plates enhance chemical biology and translation medicine in our research programs and foster new collaborative research and academic-commercial partnerships. The current laboratory infrastructure enables investigators to identify molecules that affect the activity of a target via a labeled (i.e. radioactive, fluoresent, chemiluminescent) tracer/substrate. Although readouts from labeled substrates are very sensitive and amenable to high-throughput, they are also prone to interference, leading to increased false-positive and - negative discovery rates, and provide no evidence that modulators directly bind to the target of interest. Since its inception, the laboratory has facilitted the assay development and execution of small molecule screens against 10 different targets, including gamma Herpes Virus latency, Human Papillomavirus oncoproteins, p53, and human telomerase. The results from these screens have led to the identification of 2-3 series of confirmed "hits" for each project that are ready for optimization through medicinal chemistry. However, existing technology (i.e. ITC, sedimentation equilibrium by analytical ultracentrifugation) are lower throughput and require large amounts of rare or sensitive materials to assess interactions between the small molecule and target. As such, the requested Biacore T200 will be located and maintained in the Wistar Institute Molecular Screening Facility. The sensitivity, versatility, and throughput of the Biacore T200 will immediately impact the ability of6 Major NIH-funded users and a group of Minor users to quantitatively evaluate a broad spectrum of biomolecular interactions and develop Fragment Based Screening, as an alternative to small molecule inhibitors of protein function. An internal advisory committee comprised of users and senior staff from the Wistar Institute will oversee the operation, maintenance, and equitable access to this instrument. In this application, we present projects and preliminary data from studies that require access to sensitivity of Surface Plasmon Resonance label-free detection. The purchase of the Biacore T200 will greatly enhance the ability of investigators to characterize biomolecular interactions and expand the capabilities and versatility of the Wistar Molecular Screening Facility to support transformative research aimed to reduce the burden of human disease.