Bruker Sierra SPR-24 Pro

项目摘要/摘要 此应用程序申请资金购买高通量表面等离子体激元Sierra SPR-24 Pro(Bruker) 共振生物传感器(HT-SPR)。该仪器将取代已过时的Biacore 4000(Cytiva) 已经运作了九年多。SPR是一种非常灵敏的测量结合相互作用的方法 在固定化分子和溶液相分子之间进行实时检测。测量到的信号是 与质量变化成正比，允许确定结合化学计量比以及结合/解除结合 动力学和平衡结合常数。该仪器将放置在加州大学旧金山分校的小分子发现号上 中心(SMDC)，高通量筛选、化学生物学和药物的最新核心设施 发现号。19名调查人员将在头两年使用该仪器。美国国立卫生研究院资助的九个主要用户 要求仪器立即表征蛋白质-蛋白质和蛋白质/小分子相互作用 去发现新的探测器。生物学领域包括：1)蛋白质稳态-AAA-ATPase在癌症和罕见疾病中的作用 疾病，癌症和神经变性中的伴侣，病毒学中的泛素连接酶；2)调节酶- 癌症中的去甲基酶，免疫肿瘤学中的nSMase；3)冠状病毒抗体和设计的蛋白质为 药物和诊断；4)膜蛋白--癌症中的胞外蛋白酶，疾病中的铁调节。十 次要用户提供了额外的多样性，并代表了未来的潜在主要用户。SPR-24将取代 已有9年历史的Biacore 4000，现在已经过时，不能用于当前和预期的用途。具体地说，1)仪器 不再受Cytiva支持，关键部件不再可用；2)该系统不适合小型项目， 导致SMDC拒绝了几个用户，以及3)数据管理和数据分析软件 对大型项目不方便，导致花费大量时间将文件传输到第三方软件。这个 不同的研究项目要求SPR仪器灵敏、坚固，并允许灵活地 实验设计。我们研究了四种仪器；虽然所有仪器都足够灵敏，可以满足 项目，我们确定SPR-24包括几个理想的功能。首先，芯片架构提供 在三个表面上同时进行八次注射，可实现高度的多路复用；此外，样品 可通过灵活的用户界面从板材中选择，这方便了小型实验。的成本 该仪器非常优惠，使用成本也很有竞争力。最后，该仪器节省了空间。 现在和以后可以自动化；它具有容纳两个384孔板的内部容量，并可以连接到机器人 板材处理器，将购买，如果未来的项目需要它。SMDC拥有13年的历史记录， 使用已建立的基于网络的日历和充值系统成功管理高端设备。一个 专家生物物理学家将得到该机构的支持，确保仪器完好无损 维护后，用户将得到专业培训，19名调查人员将获得关键数据。

PROJECT SUMMARY/ABSTRACT This application requests funds to purchase a Sierra SPR-24 Pro (Bruker), a high-throughput surface plasmon resonance biosensor (HT-SPR). The instrument will replace a deprecated Biacore 4000 (Cytiva) that has been in operation for over nine years. SPR is an exquisitely sensitive method for measuring binding interactions between an immobilized molecule and a solution-phase molecule in real time. The measured signal is proportional to the change in mass, allowing determination of binding stoichiometry as well as binding/unbinding kinetics and equilibrium binding constants. The instrument will be placed in the UCSF Small Molecule Discovery Center (SMDC), a state-of-the-art core facility for high-throughput screening, chemical biology, and drug discovery. Nineteen investigators will use the instrument over the first two years. Nine NIH-funded major users require the instrument immediately to characterize protein-protein and protein/small-molecule interactions and to discover new probes. Areas of biology include: 1) protein homeostasis - AAA+ ATPases in cancer and rare disease, chaperones in cancer and neurodegeneration, ubiquitin ligases in virology; 2) regulatory enzymes – demethylases in cancer, nSMase in immuno-oncology; 3) coronavirus – antibodies and designed proteins as drugs and diagnostics; 4) membrane proteins - extracellular proteases in cancer, iron regulation in disease. Ten minor users provide additional diversity and represent potential major users in the future. The SPR-24 will replace a 9-year old Biacore 4000 that is now obsolete for current and anticipated usage. Specifically, 1) the instrument is no longer supported by Cytiva and key parts are no longer available, 2) the system is ill-suited to small projects, leading the SMDC to turn away several users, and 3) data management and data analysis software are inconvenient for large projects, resulting in significant time spent transferring files to third-party software. The diverse research projects require SPR instrumentation that is sensitive, robust, and allows flexibility in experimental design. We researched four instruments; while all are sensitive enough for the needs of the projects, we determined that the SPR-24 included several ideal features. First, the chip architecture provides eight simultaneous injections across three surfaces, allowing a high degree of multiplexing; additionally, samples can be selected from the plate through a flexible user interface, which facilitates small experiments. The cost of the instrument is highly favorable and the cost-of-use is competitive. Finally, the instrument is space efficient now and automatable later; it has internal capacity to hold two 384-well plates and can be connected to a robotic plate handler that will be purchased if future projects require it. The SMDC has a thirteen-year track record of successfully managing high-end equipment using an established web-based calendar and recharge system. An expert biophysical scientist will be supported by the institution, ensuring that the instrument will be well maintained, users will be expertly trained, and the nineteen investigators will obtain their critical data.