ThermoFisher Scientific Orbitrap Eclipse with ETD and UVPD

项目概要： 这项修订后的提案寻求资金，以获得一个热轨道阱日食质谱仪 配备ETD和UVPD，以满足HDX-MS和糖蛋白组学研究的关键需求， 华盛顿大学。这个系统将使我们的团队能够克服技术限制， 和样本分析瓶颈，阻碍和停滞了许多NIH资助的项目。讣告 Eclipse平台是扩展我们的项目范围和吞吐量的最合适的平台。 HDX-MS研究：1）提供最先进的灵敏度，检测下限，使HDX- MS研究非常有限的材料样本，包括完整的病毒颗粒和难以纯化的蛋白质 2）达到500，000的质量分辨率，这是天然生物来源的10倍。 可在我们现有的仪器上获得，并且是分析高度复杂样品所必需的; 3） 能够进行常规和稳健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）紫外线吸收 光解离（UVPD），用于扩展可从HDX-MS研究获得的分辨率;和5） 能够通过ETD和UVPD对经典难治性蛋白质进行自上而下的HDX-MS研究 自下而上的方法。在迅速的时间范围内将联合收割机不同的破碎方法结合起来的能力将 能够开发用于位点特异性糖蛋白质组学研究的新工具， 糖生物学的聚糖结构解析所需的工具。这一工具，以其广泛的 能力，也将有利于一般蛋白质组学的应用和鉴定罕见的和新的后， 翻译修饰虽然有几个质谱仪可在华盛顿大学，所有的高端 系统被大量利用并专用于其它应用。HDX-MS系统是 我们的大多数用户都住在药学院质谱中心，不能 移动并与设施外的仪器连接。此外，没有一个单一的 UVPD使高分辨率质谱仪在华盛顿大学或任何地方在更大的西雅图地区， 满足当前的技术需求。这一共同工具将提供能力和途径， 需要推进目前NIH资助的研究，并将使我们能够推动蛋白质的边界 HDX-MS分析和生物医学研究中的位点特异性糖蛋白质组学研究，以支持未来 NIH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.