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.

项目概要： 该修订后的提案寻求资金购买 Thermo Orbitrap Eclipse 质谱仪 配备 ETD 和 UVPD，可满足 HDX-MS 和糖蛋白组学研究的关键需求 华盛顿大学。该系统将使我们的团队克服技术限制 样本分析瓶颈阻碍并阻碍了许多 NIH 资助的项目。讣告 Eclipse 平台是最适合扩大我们的项目范围和吞吐量的平台 HDX-MS 研究如下： 1) 提供最先进的灵敏度以降低检测限，以实现 HDX- 对材料非常有限的样品进行质谱研究，包括完整的病毒颗粒和难以纯化的蛋白质 来自天然生物来源； 2) 质量分辨率达到500,000，比传统技术高十倍 可在我们当前的仪器上获得，并且对于分析高度复杂的样品是必要的； 3) 是 能够进行常规且稳定的高通量电子转移解离（ETD），这对于 复杂蛋白质样品和糖肽的鉴定； 4) 加入紫外线 光解离 (UVPD) 用于扩大 HDX-MS 研究获得的分辨率；和 5) 通过 ETD 和 UVPD 对经典难降解的蛋白质进行自上而下的 HDX-MS 研究 自下而上的方法。快速组合不同分段方法的能力将 能够开发用于位点特异性糖蛋白组学研究的新工具和许多 糖生物学聚糖结构阐明所需的工具。该仪器以其广泛的 能力，也将有利于一般蛋白质组学应用和稀有和新颖的后蛋白质的鉴定 翻译修饰。虽然威斯康星大学提供多种质谱仪，但所有高端 系统被大量使用并致力于其他应用程序。所需的 HDX-MS 系统 我们的大多数用户都位于药学院质谱中心，无法 可以移动并与设施外部的仪器连接。此外，没有一个单一的 UVPD 在华盛顿大学或大西雅图地区的任何地方启用了高分辨率质谱仪 支持当前的技术需求。该共享工具将提供功能和访问 需要推进当前 NIH 资助的研究，并使我们能够突破蛋白质的界限 通过 HDX-MS 进行分析以及生物医学研究中的位点特异性糖蛋白组学研究以支持未来 NIH 资助的项目。