Thermo Scientific Orbitrap Eclipse Tribrid ETD Mass Spectrometer for UConn Proteomics

项目摘要/摘要 这项建议的目的是获得资金购买新的Thermo Science Orbitrap月食三合一 支持ETD的质谱仪和完全专用的Dionex旗舰3000 RSLCNano超高性能 康涅狄格大学新的尖端蛋白质组学分析的高效液相色谱系统 (康涅狄格州)。拟议的仪器将是康涅狄格州大学蛋白质组学和代谢组学的组成部分 位于斯托尔斯学院校园的设施(PMF)。PMF是基于质谱学(MS)的共享 为整个康涅狄格州大学研究社区提供创新和定制方法的资源 多肽、蛋白质和小分子代谢物的鉴定和定量，以阐明 人类疾病背后的分子机制。自2.5年前开业以来，PMF已经提供了 为80多个实验室提供复杂的分析服务，其中许多实验室有多个项目 同时进行。目前，康涅狄格州大学的所有蛋白质组学实验都是在PMF中使用单个、 2015年购买的翻新Thermo Science Q Exactive HF质谱仪；这是唯一高 康涅狄格州大学调查人员可使用分辨率先进的蛋白质组学质谱仪。因此，它 始终满负荷运行，缺乏NIH资助的研究所需的关键分析选择。这个 所要求的仪器将独家提供健康所需的实质性技术升级- 相关项目侧重于阐明蛋白质对各种人类疾病的贡献。这些至关重要的 升级包括对低丰度离子检测的最高灵敏度，卓越的蛋白质组覆盖深度 对于每个实验，康涅狄格州大学没有新的肽/蛋白质解离技术，以及新的方法来 通过实时搜索为基于标记的多路蛋白质组学提供最高精度的量化 同步前身选择。这些功能在建议的仪器上独一无二地展示， 康涅狄格州大学斯托尔斯分校和康涅狄格州大学健康医学研究中心的15名(13名由NIH资助)研究人员需要 校园。相反，我们现有的单一仪器灵敏度较低，覆盖深度有限。 特别是对于蛋白质有限的样品，串联质谱仪仅用一种多肽解离技术，且不准确 基于标签的多路分析的量化。因此，收购Orbitrap Eclipse是必要的 对我们目前无法成功完成的NIH资助项目的科学研究进行转化。通过 实现Orbitrap Eclipse上的新技术，PMF将允许进行突破性研究 需要分析严重有限的蛋白质样本，全面表征重要的监管后 翻译修饰和识别与疾病有关的新的蛋白质靶点 准确的定量蛋白质组学。总体而言，这些进步将使NIH资助的康涅狄格州大学调查人员能够 确定潜在的蛋白质对人类疾病的贡献，并帮助设计未来的治疗方法。

Project Summary/Abstract The purpose of this proposal is to acquire funds to purchase a new Thermo Scientific Orbitrap Eclipse Tribrid ETD-enabled mass spectrometer and fully dedicated Dionex Ultimate 3000 RSLCnano ultra-high performance liquid chromatography system to enable new, cutting-edge proteomics analysis at the University of Connecticut (UConn). The proposed instrumentation will be an integral part of the UConn Proteomics and Metabolomics Facility (PMF) located on the Storrs academic campus. PMF is a mass spectrometry (MS)-based shared resource that provides the entire UConn research community with innovative and customized methods for the identification and quantification of peptides, proteins, and small molecule metabolites in order to elucidate the molecular mechanisms that underlie human disease. Since opening 2.5 years ago, PMF has already provided sophisticated analytical services to more than 80 laboratories and many of these have multiple projects ongoing simultaneously. Currently, all proteomics experiments at UConn are analyzed in PMF using a single, refurbished Thermo Scientific Q Exactive HF mass spectrometer purchased in 2015; this is the only high resolution advanced proteomics-capable mass spectrometer available to UConn investigators. As a result, it consistently runs at full capacity and lacks crucial analytical options required for NIH-funded research. The requested instrumentation will exclusively provide the substantial technological upgrades required for health- related projects focused on elucidating the protein contributions to various human diseases. These vital upgrades include maximum sensitivity for low abundance ion detection, superior depth of proteome coverage for every experiment, new peptide/protein dissociation techniques unavailable at UConn, and novel methods to provide the utmost accuracy of quantification for label-based, multiplexed proteomics via Real-Time Search Synchronous Precursor Selection. These features are uniquely exhibited on the proposed instrumentation and are required for 15 (13 NIH-funded) investigators at the UConn Storrs and UConn Health medical research campuses. Conversely, our single existing instrument suffers from poorer sensitivity, limited depth of coverage particularly for protein-limited samples, tandem MS by only one peptide dissociation technique, and inaccurate quantification for label-based, multiplexed analyses. Therefore, acquisition of the Orbitrap Eclipse is necessary to transform scientific research for NIH-funded projects that we cannot currently complete successfully. By implementing new technologies featured on the Orbitrap Eclipse, PMF will permit breakthrough research that requires analysis of severely limited protein samples, full characterization of important regulatory post- translational modifications, and identification of new protein targets implicated in disease using extremely accurate quantitative proteomics. Overall, these advancements will allow NIH-funded UConn investigators to determine the underlying protein contributions to human disease and help design future therapeutics.