Bruker Sierra SPR-24 Pro

项目总结/摘要 该申请要求资金购买Sierra SPR-24 Pro（Bruker），一种高通量表面等离子体激元 共振生物传感器（HT-SPR）。该仪器将取代已弃用的Biacore 4000（Cytiva）， 运行了9年多。SPR是一种非常灵敏的测量结合相互作用的方法 固定化分子和溶液相分子之间的真实的时间。所测量的信号 与质量变化成比例，允许确定结合化学计量以及结合/未结合 动力学和平衡结合常数。该仪器将被放置在UCSF小分子发现 中心（SMDC），高通量筛选，化学生物学和药物的最先进的核心设施 的发现19名研究人员将在头两年使用该仪器。九个NIH资助的主要用户 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， 来发现新的探测器生物学领域包括：1）蛋白质稳态- AAA+ ATP酶在癌症和罕见 疾病，癌症和神经变性中的伴侣蛋白，病毒学中的泛素连接酶; 2）调节酶- 癌症中的脱甲基酶，免疫肿瘤学中的nSMase; 3）冠状病毒-抗体和设计的蛋白质， 药物和诊断; 4）膜蛋白-癌症中的细胞外蛋白酶，疾病中的铁调节。十 次要用户提供了额外的多样性，并代表未来的潜在主要用户。SPR-24将取代 9年前的Biacore 4000，现在已经过时，不适合当前和预期的使用。具体而言，（1）工具 Cytiva不再支持，关键部件不再可用，2）该系统不适合小型项目， 导致SMDC拒绝了一些用户，3）数据管理和数据分析软件， 不适合大型项目，导致将文件传输到第三方软件花费大量时间。的 不同的研究项目需要SPR仪器，该仪器灵敏、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， 项目中，我们确定SPR-24包括几个理想的功能。首先，芯片架构提供了 在三个表面上同时进样八次，允许高度复用;此外， 可以通过灵活的用户界面从板中选择，这有利于小型实验。的成本 该仪器是非常有利的，并且使用成本是有竞争力的。最后，该仪器具有空间效率 现在和自动化以后;它有内部容量，以容纳两个384孔板，并可以连接到一个机器人 如果未来的项目需要，将购买板处理器。SMDC有13年的跟踪记录， 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 专家生物物理科学家将由该机构的支持，确保该仪器将很好地 用户将接受专业培训，19名调查员将获得关键数据。

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.