ThermoFisher Scientific Orbitrap Eclipse with ETD and UVPD

项目总结： 这项修订后的提案寻求资金，以购买Thermo Orbitrap日食质谱仪 配备ETD和UVPD，以满足HDX-MS和糖蛋白质组研究的关键需求 华盛顿大学。这个系统将允许我们的团队克服技术限制 以及样本分析瓶颈，这些瓶颈阻碍和拖延了许多NIH资助的项目。Obitrap ECLIPSE平台是最适合于扩大我们的项目范围和吞吐量的平台 HDX-MS的研究如下：1)提供最先进的检测下限灵敏度，以使HDX-MS能够- 材料非常有限的样品的MS研究，包括完整的病毒颗粒和难以纯化的蛋白质 来自本地生物来源；2)达到50万的质量分辨率，这比 在我们目前的仪器上可以获得，对于分析高度复杂的样品是必要的；3)是 能够常规和强大的高通量电子转移解离(ETD)，这对 复合蛋白质样品和糖肽的鉴定；4)结合紫外光 用于扩展从HDX-MS研究获得的分辨率的光解离(UVPD)；以及5) 通过ETD和UVPD实现对经典蛋白质的自上而下的HDX-MS研究 自下而上的方法。能够将不同的分段方法组合在一个快速的时间尺度上 能够开发用于位点特异性糖蛋白组学研究的新工具和开发多个 糖生物学所需的糖链结构阐明工具。这件乐器，带着它的宽阔 能力，也将有利于一般蛋白质组学的应用和识别稀有和新的后 翻译修饰语。虽然威斯康星大学提供了几种质谱仪，但所有高端 系统被大量利用，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在药学院质谱学中心，不能 可移动到设施外的仪器并与其对接。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。该共享仪器将提供以下功能和访问 以推进目前由美国国立卫生研究院资助的研究，并将使我们能够突破蛋白质的界限 生物医学研究中的HDX-MS分析和定点糖蛋白质组研究为未来提供支持 美国国立卫生研究院资助的项目。

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.