Acquisition of a Dual-Source, High-Performance, Ion Mobility, Quadrupole Time-of-Flight Mass Spectrometry System for Biomedical Research at UW-Madison

项目总结/摘要 威斯康星大学麦迪逊分校的前沿研究导致了定量和 研究性蛋白质组学，功能性糖组学询问，生物活性分子的结构表征， 天然产物发现和代谢组学分析，重点关注人类健康和疾病。后 评估26名NIH支持的研究人员的当前需求，并尽职地比较商业高， 分辨率质谱仪产品，该提案寻求资金购买仪器能力 减轻分析灵敏度、质量分辨率、离子分离和占空比方面的限制， 目前可获得的光谱仪，并直接受益于每个研究重点。考虑到 人类和疾病相关样品，电喷雾（ESI）和基质辅助激光解吸电离 基于MALDI的质谱（MS）仪器显示出由于有限的干扰而导致的灵敏度的限制。 前体选择、来自单电荷离子的干扰以及可消除高达90%离子的低占空比 在分析之前。在最初的选择中幸存下来的物种，通过串联识别生物分子， MS扫描可能会受到前体共分离和低丰度分析物信号抑制的严重阻碍。 考虑到这些限制，新兴的文献和个人评价表明，被困离子迁移率 光谱法（TIMS）是唯一能够扩展蛋白质组学中的分析灵敏度的范例， 糖蛋白质组学和小分子分析，同时还提供了最高的气相分辨率制度， 结构和构象研究。布鲁克timsTOF flexX MS的创新仪器设计 系统结合了几个新颖的设计改进的源极区，TIMS细胞，和四极， 显著提高仪器性能。我们对timsTOF flexX的初步测试以及与其他 目前的高分辨率轨道阱和离子迁移率仪器表明，获得timsTOF fleX提供了 对我们的研究合作伙伴最大的好处，因为它是蛋白质组学，糖蛋白质组学， 代谢组学和离子迁移率分析。这种新仪器将提供先进的质谱功能，以支持 26名高生产力的NIH资助的研究人员与73个正在进行的项目的研究。在这一广泛领域取得的进展 将通过与各组织的密切合作，有效利用新工具， 用户群，李博士，一个高生产力的教员超过27年的经验，生物质量 和Scarlett博士，UW-Madison Pharmacy-MS设施主任。替代文书或 如果这项申请获得拨款，当局定会考虑拨款。

PROJECT SUMMARY/ABSTRACT Cutting-edge research at the University of Wisconsin-Madison has led to pioneering efforts in quantitative and investigational proteomics, functional glycomic interrogation, structural characterization of bioactive molecules, natural product discovery and metabolomic profiling with significant focus on human health and disease. Upon evaluating the current needs of 26 NIH-supported investigators and dutiful comparison of commercial high- resolution mass spectrometer offerings, this proposal seeks funding for the purchase of instrumentation capable of alleviating the limitations in analytical sensitivity, mass resolution, ion separation, and duty cycle presented by currently-accessible spectrometers and that directly benefits each research focus. Given the complexity of human and disease-relevant samples, electrospray (ESI) and matrix-assisted laser desorption ionization (MALDI)-based mass spectrometry (MS) instrumentation demonstrate limitations in sensitivity due to limited precursor selection, interference from singly-charged ions, and low duty cycles that eliminate up to 90% of ions prior to analysis. Of the species that do survive initial selection, identification of biomolecules through tandem MS scans can be severely hindered by precursor co-isolation and signal suppression of low abundance analytes. With these limitations in mind, emerging literature and personal evaluations demonstrate that trapped ion mobility spectrometry (TIMS) is the paradigm uniquely capable of expanding analytical sensitivity in proteomic, glycoproteomic, and small molecule analyses, while also presenting the highest gas-phase resolution regime for structural and conformational investigation. The innovative instrument design of the Bruker timsTOF fleX MS system incorporates several novel design improvements to the source area, TIMS cell, and quadrupole to dramatically enhance instrument performance. Our initial testing of the timsTOF fleX and comparison to other current high-resolution orbitrap and ion mobility instruments indicate the acquisition of the timsTOF fleX provides the greatest benefit to our research partners, as it is the most effective regime in proteomic, glycoproteomic, metabolomic and ion mobility analysis. This new instrument will provide advanced MS capabilities to support the research of 26 highly productive NIH-funded investigators with 73 ongoing projects. Progress on this broad array of projects will be catalyzed by the effective usage of the new instrument through close cooperation among the user groups, Dr. Li, a highly productive faculty member with more than 27 years of experience in biological mass spectrometry, and Dr. Scarlett, the UW-Madison Pharmacy-MS Facility Director. Alternative instruments or allocation of funds would certainly be considered, should this application be funded.