Bruker Sierra SPR-24 Pro

布鲁克塞拉 SPR-24 Pro

• 批准号: 10175786
• 负责人: Michelle Arkin
• 金额: $ 30.43万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10175786
• 关键词: 9 year old; ATP phosphohydrolase; Antibodies; Architecture; Area; Binding; Biology; Biophysics; Biosensor; Calendar; Chemicals; Computer software; Core Facility; Coronavirus; Data; Data Analyses; Diagnostic; Disease; Ensure; Enzymes; Equilibrium; Equipment; Experimental Designs; Extracellular Protein; Funding; Future; Immobilization; Immunooncology; Injections; Institution; Iron; Kinetics; Malignant Neoplasms; Measures; Membrane Proteins; Methods; Minor; Molecular Chaperones; Nerve Degeneration; Online Systems; Peptide Hydrolases; Pharmaceutical Preparations; Phase; Protein Engineering; Proteins; Rare Diseases; Regulation; Request for Applications; Research; Research Personnel; Research Project Grants; Robotics; Sampling; Scientist; Signal Transduction; Surface; Surface Plasmon Resonance; System; Time; Training; United States National Institutes of Health; cost; data management; drug discovery; experimental study; flexibility; high throughput screening; instrument; instrumentation; operation; proteostasis; small molecule; stoichiometry; ubiquitin ligase; virology

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.

项目摘要/摘要 该申请要求资金购买Sierra Spr-24 Pro（Bruker），这是一个高通量表面的高素质 共振生物传感器（HT-SPR）。该仪器将取代已弃用的biacore 4000（Cytiva） 在运行九年以上。 SPR是测量结合相互作用的精确敏感方法 实时在固定分子和溶液相分子之间。测得的信号是 与质量变化成正比，允许确定结合化的化学计量和结合/解除 动力学和平衡结合常数。该仪器将放置在UCSF小分子发现中 中心（SMDC），一种用于高通量筛查，化学生物学和药物的最先进的核心设施 发现。十九个调查人员将在头两年使用该工具。 NIH资助的主要用户 需要立即该仪器表征蛋白质 - 蛋白质和蛋白质/小分子相互作用，以及 发现新问题。生物学领域包括：1）蛋白质稳态 - 癌症和罕见的AAA+ ATPase 疾病，癌症和神经退行性的伴侣，病毒学中的泛素连接酶； 2）调节酶 - 癌症中的脱甲基酶，免疫肿瘤中的NSMase； 3）冠状病毒 - 抗体和设计蛋白 药物和诊断； 4）膜蛋白 - 癌症中的细胞外蛋白，疾病中的铁调节。十 次要用户提供额外的多样性，并代表未来潜在的主要用户。 SPR-24将替换 现年9岁的Biacore 4000现在已经过时使用了当前和预期的用法。具体而言，1）仪器 不再受细胞胞菌的支持，关键部分不再可用，2）该系统不适合小型项目， 带领SMDC拒绝了几个用户，3）数据管理和数据分析软件是 大型项目的不便，导致将文件传输到第三方软件花费了大量时间。这 多样化的研究项目需要SPR仪器敏感，稳健，并允许灵活性 实验设计。我们研究了四种乐器。虽然所有这些都足够敏感 项目，我们确定SPR-24包括几个理想功能。首先，芯片架构提供 在三个表面上进行了八次简单注射，可以高度多重。另外，样品 可以通过灵活的用户界面从板上选择该设施小实验。费用 该工具非常有利，使用成本具有竞争力。最后，该乐器是有效的 现在，以后可自动；它具有内部容纳两个384孔板的能力，可以连接到机器人 如果将来的项目需要它，将购买的板厂处理程序。 SMDC拥有十三年的记录 使用已建立的基于Web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得到该机构的支持，以确保该工具会很好 维护，用户将接受专业培训，而十九个调查人员将获得其关键数据。