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

不断改进高度复杂的样品的蛋白质组学特征，如完整蛋白质 对质谱仪的性能提出了更高的要求。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 都需要提供临床医生和研究人员产生主要影响所需的数据 关于所提议的八个DBP中的每一个关于人类健康的设想。以质量分析为基础 作为蛋白质组学的核心，这些创新对拟议的DBP的影响很难被低估。分成两份 具体目标，我们提出了两个新的工具。第一个是改进的Orbitrap融合仪，减少了 真空压力和紫外准分子激光193 nm光解的加入。第二 仪器建立在Exactive平台上，增加了一个飞行时间分离器来取代四极杆 高效率、并行隔离和动态范围扩大，可同时分析更多离子。 DBP提出的资源需求专门为开发新的仪器而量身定做 在样本有限的应用中，整体蛋白质分析面临的挑战。新的快速碎片化 方法，结合选择性积累和已经令人印象深刻的碎片化工具箱，将产生 在彻底确定蛋白质靶标的能力方面无与伦比的仪器(基于Orbitrap Fusion 仪器)。作为这些能力的补充，TOF-Orbitrap具有并行选择和 动态范围增强将发现和识别前所未见的蛋白质。它们之间的耦合 具有在研发1中描述的分离方法的仪器将产生分析的基础 为DBP合作者获取最佳可能结果所需的平台。

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.