Next-Generation Fourier-Transform Mass Spectrometers for Interrogation of Whole Proteins (pp 481-522)

不断改进高度复杂样品（例如完整蛋白质）的蛋白质组表征 对质谱仪器提出了更高的性能要求。改进检测的创新 完整蛋白质，添加新的和不同的碎片模式，增加动态范围并减少扫描时间 都需要让临床医生和研究人员能够获取产生重大影响所需的数据 拟议的八项 DBP 中的每一项都涉及人类健康。随着质量分析 作为蛋白质组学的核心，这些创新对拟议的 DBP 的影响很难低估。分成两份 为了具体目标，我们提出了两种新文书。第一个是改进的 Orbitrap Fusion 仪器，减少了 真空压力和添加用于 193nm 光解的紫外准分子激光器。第二个 仪器建立在 Exactive 平台之上，添加了飞行时间分离器来代替四极杆 高效、并行隔离和动态范围增强，可同时分析更多离子。 资源提出的 DBP 要求开发专门定制的新仪器 应对样品有限的应用中全蛋白质分析的挑战。新的、快速的碎片化 方法，结合选择性积累和已经令人印象深刻的碎片工具箱，将产生 一种能够彻底表征蛋白质目标的无与伦比的仪器（基于 Orbitrap Fusion 仪器）。作为这些功能的补充，TOF-Orbitrap 具有并行选择和 动态范围增强将发现和识别以前从未见过的蛋白质。这些的耦合 采用 TR&D 1 中描述的分离方法的仪器将为分析奠定基础 为 DBP 合作者获取最佳结果所需的平台。

Improved proteomic characterization of highly complex samples, such as intact proteins, continually demands higher performance from mass spectrometric instrumentation. Innovations that improve the detection of intact proteins, add new and different fragmentation modalities, increase dynamic range and reduce scan time are all required to make accessible the data needed by clinicians and researchers to generate the major impacts on human health envisaged in each and every one of the eight DBPs proposed. With mass analysis being central to proteomics, the impact of these innovations on the proposed DBPs is difficult to understate. In two specific aims, we present two new instruments. The first is a modified Orbitrap Fusion instrument with reduced vacuum pressure and the addition of an ultraviolet excimer laser for 193nm photodissociation. The second instrument builds upon the Exactive platform, adding a time-of-flight separator in place of the quadrupole for high-efficiency, parallel isolation and dynamic range enhancement to analyze more ions simultaneously. The DBPs proposed by the resource demand the development of new instrumentation specifically tailored to the challenges of whole protein analysis in applications where sample is limited. New and fast fragmentation approaches, combined with selective accumulation and an already impressive fragmentation tool box, will yield an instrument unsurpassed in the ability to thoroughly characterize protein targets (Orbitrap Fusion-based instrument). Functioning as a complement to these capabilities, a TOF-Orbitrap, with parallel selection and dynamic range enhancement will uncover and identify proteins never before seen. The coupling of these instruments with the separation approaches described in TR&D 1 will produce the foundation of the analytical platform required to extract the best possible outcomes for the DBP collaborators.