Thermo Scientific Orbitrap Eclipse Tribrid ETD Mass Spectrometer for UConn Proteomics

项目总结/摘要 该提案的目的是获得资金，以购买新的Thermo Scientific Orbitrap Eclipse Tribrid 支持ETD的质谱仪和完全专用的Dionex Ultimate 3000 RSLCnano超高性能 液相色谱系统，使新的，尖端的蛋白质组学分析在康涅狄格大学 （UConn）.拟议的仪器将是康州大学蛋白质组学和代谢组学的一个组成部分 设施（PMF）位于斯托尔斯学术校园。PMF是基于质谱（MS）的共享 资源，为整个康州大学研究界提供创新和定制的方法， 肽、蛋白质和小分子代谢物的鉴定和定量，以阐明 人类疾病的分子机制自2.5年前开业以来，PMF已经提供 为80多个实验室提供先进的分析服务，其中许多实验室都有多个项目 同时进行。目前，在康州大学的所有蛋白质组学实验都是在PMF中使用单一的， 2015年购买的翻新Thermo Scientific Q Exactive HF质谱仪;这是唯一一台 分辨率先进的蛋白质组学能力的质谱仪提供给康州大学的研究人员。结果它 它一直在满负荷运行，缺乏NIH资助的研究所需的关键分析选项。的 所要求的仪器将专门提供卫生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作用。这些重要 升级包括低丰度离子检测的最大灵敏度，蛋白质组覆盖的上级深度 对于每一个实验，新的肽/蛋白质解离技术在康州大学不可用，和新的方法， 通过实时搜索为基于标记的多重蛋白质组学提供最高的定量准确度 同步前体选择。这些特征在申报手术工具上表现独特， 康州大学斯托尔斯和康州大学健康医学研究所的15名（13名NIH资助的）研究人员需要 校园。相反，我们现有的单一仪器灵敏度较差，覆盖深度有限， 特别是对于蛋白质有限的样品，串联MS仅通过一种肽解离技术， 用于基于标记的多重分析的定量。因此，收购Orbitrap Eclipse是必要的 为我们目前无法成功完成的NIH资助项目转变科学研究。通过 实施Orbitrap Eclipse上的新技术，PMF将允许突破性的研究， 需要分析非常有限的蛋白质样品，充分表征重要的调控后 翻译修饰，并使用极端的方法鉴定与疾病有关的新蛋白质靶点。 精确的定量蛋白质组学。总的来说，这些进步将使NIH资助的康州大学研究人员能够 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.