ThermoFisher Scientific Orbitrap Eclipse with ETD and UVPD

带 ETD 和 UVPD 的 ThermoFisher Scientific Orbitrap Eclipse

• 批准号: 10427682
• 负责人: Miklos Guttman
• 金额: $ 118.42万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10427682
• 关键词: Address; Area; Biological; Biomedical Research; Complex; Development; Dissociation; Electron Transport; Funding; Future; Glycobiology; Glycopeptides; Mass Spectrum Analysis; Methods; Pharmacy Schools; Polysaccharides; Post-Translational Protein Processing; Protein Analysis; Proteins; Proteomics; Refractory; Research; Resolution; Sampling; Site; Source; System; Time; United States National Institutes of Health; Universities; Virion; Washington; detection limit; glycoproteomics; instrument; instrumentation; mass spectrometer; novel; protein complex; tool; ultraviolet

Project summary: This revised proposal seeks funds to acquire a Thermo Orbitrap Eclipse mass spectrometer equipped with ETD and UVPD to address critical needs for HDX-MS and glycoproteomics studies at the University of Washington. This system would allow our groups to overcome the technical limitations and sample analysis bottlenecks that have hindered and stalled many NIH funded projects. The Obitrap Eclipse platform is the most suitable platform for expanding the projects scopes and throughput of our HDX-MS studies as it: 1) Offers state of the art sensitivity to lower limits of detection to enable HDX- MS studies of very material-limited samples, including intact virus particles and hard to purify proteins from native biological sources; 2) Achieves a mass resolution of 500,000 which is ten times higher than obtainable on our current instrumentation and is necessary for analyzing highly complex samples; 3) Is capable of routine and robust high-throughput electron transfer dissociation (ETD), which is vital for identification of complex protein samples and glycopeptides; 4) Incorporates ultra-violet photodissociation (UVPD) for expanding the resolution obtainable from HDX-MS studies; and 5) Enables top-down HDX-MS studies through both ETD and UVPD for proteins refractory to classical bottom-up approaches. The capacity to combine different fragmentation methods a rapid timescale will enable development of new tools for site-specific glycoproteomics studies and development of much needed tools for glycan structural elucidation for glycobiology. This instrument, with its broad capabilities, will also benefit general proteomics applications and identification of rare and novel post- translational modifications. While several mass spectrometers are available at UW, all the high-end systems are heavily utilized and devoted to other applications. The HDX-MS system that is required for the majority of our users is housed in the School of Pharmacy Mass Spectrometry Center and cannot be moved and interfaced with the instruments outside of the facility. Additionally, there is not a single UVPD enabled high resolution mass spectrometer at UW or anywhere in the greater Seattle area to support out current technological needs. This shared instrument will provide the capabilities and access needed to advance current NIH funded research and will allow us to push the boundaries of protein analysis by HDX-MS and site-specific glycoproteomics studies in biomedical research to support future NIH funded projects.

项目摘要： 该修订的提案寻求资金来获取热轨道日食质谱仪 配备ETD和UVPD，以满足HDX-MS和糖蛋白质组学研究的关键需求 华盛顿大学。该系统将允许我们的小组克服技术限制 样本分析瓶颈阻碍了许多NIH资助的项目。 itrap Eclipse平台是扩展项目范围和我们的吞吐量的最合适的平台 HDX-MS的研究：1）对检测下限的最高敏感性，以实现HDX- MS研究非常有限的样品，包括完整的病毒颗粒和难以纯化蛋白质 来自本地生物学来源； 2）达到500,000的质量分辨率，比 在我们当前的仪器上可获得，对于分析高度复杂的样品是必不可少的； 3）是 能够常规和强大的高通量电子传递解离（ETD），这对于 鉴定复杂的蛋白质样品和糖肽； 4）融合了超紫色 用于扩大可从HDX-MS研究获得的分辨率的光解离（UVPD）； 5） 通过ETD和UVPD启用自上而下的HDX-MS研究，以使蛋白质难治性 自下而上的方法。结合不同分裂方法的能力快速时间尺度将 启用用于特定地点糖蛋白质组学研究的新工具，并开发了很多 需要用于糖生物学的聚糖结构阐明所需的工具。这个乐器，其广泛 能力，还将受益于一般蛋白质组学的应用，并确定稀有和新颖的后 翻译修改。虽然UW上有几个质谱仪，但所有高端 系统大量利用并专用于其他应用程序。 HDX-MS系统需要 我们的大多数用户都位于药学院质谱中心，不能 被移动并与设施外的乐器交织。另外，没有一个 UVPD启用了UW的高分辨率质谱仪或大西雅图地区的任何地方 支持当前的技术需求。这种共享工具将提供功能和访问权限 需要推进当前的NIH资助研究，并使我们能够突破蛋白质的界限 HDX-MS和生物医学研究中特定地点糖蛋白质组学研究的分析以支持未来 NIH资助的项目。