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.

项目概要/摘要 此申请请求资金购买 Sierra SPR-24 Pro (Bruker)，这是一种高通量表面等离子体激元 该仪器将取代已弃用的 Biacore 4000 (Cytiva)。 SPR 已运行超过九年，是一种极其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实时测量信号为 与质量变化成正比，可以确定结合化学计量以及结合/解除结合 该仪器将放置在 UCSF Small Molecule Discovery 中。 中心（SMDC），最先进的高通量筛选、化学生物学和药物核心设施 十九名研究人员将在头两年使用该仪器，九名 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，并且 发现新的生物学领域包括：1) 蛋白质稳态 - 癌症和罕见疾病中的 AAA+ ATP 酶。 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 – 癌症中的去甲基化酶、免疫肿瘤学中的 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酶，疾病中的铁调节。 次要用户提供了额外的多样性，并代表了未来潜在的主要用户，SPR-24 将取代。 已使用 9 年的 Biacore 4000 对于当前和预期的使用而言现已过时 具体来说，1) 该仪器。 Cytiva 不再支持且关键部件不再可用，2) 该系统不适合小型项目， 导致 SMDC 拒绝多名用户，以及 3) 数据管理和数据分析软件 对于大型项目来说不方便，导致将文件传输到第三方软件需要花费大量时间。 不同的研究项目需要灵敏、稳健且具有灵活性的 SPR 仪器 我们研究了四种仪器；所有仪器都足够灵敏，可以满足需要。 项目中，我们确定 SPR-24 包含几个理想的特性：首先，芯片架构提供了。 三个表面同时进行八次注射，可实现高度的多重样品分析； 可以通过灵活的用户界面从板中进行选择，这有利于小型实验的成本。 该仪器非常受欢迎，并且使用成本具有竞争力。最后，该仪器节省空间。 现在并且以后可以自动化；它具有容纳两个 384 孔板的内部容量，并且可以连接到机器人 如果未来的项目需要的话，SMDC 拥有 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。 专家生物物理科学家将得到该机构的支持，确保仪器良好 维护后，用户将接受专业培训，十九名调查员将获得他们的关键数据。