Nano-UPLC LTQ-Orbitrap Velos MS System for Yale University Keck Laboratory

描述(由申请人提供)：申请资金用于购买Waters NanACQUITY超性能LC和Thermo Fisher科学纳米电喷雾电离线性四极子离子陷阱-轨道捕获(LTQ-Orbitrap)Velos质谱仪系统，该系统将为耶鲁大学凯克实验室的MS和蛋白质组学资源提供独特的高性能质谱仪。LTQ-Orbitrap Velos是同类最新的Orbitrap MS平台之一，具有百万分之几的质量精度、新的大气压界面透镜设计以及其他高压离子捕获和离子传输特性，这些特性共同使新的Velos平台的速度比其Model XL前身快2倍，灵敏度高5-10倍。目前，耶鲁大学有一个LTQ-Orbitrap XL，它是在服务收费的基础上提供的，位于Keck实验室的MS和蛋白质组学资源中。该资源每年向90个机构的约100名耶鲁大学研究人员和110名非耶鲁大学研究人员提供蛋白质组学技术。对其LTQ-Orbitrap XL支持的技术的需求增长如此之快，以至于SILAC、无标记定量、磷酸蛋白质组学和许多其他依赖于LTQ-Orbitrap Velos的技术的延迟现在已经达到&gt；75天，该仪器以最大容量连续运行。将所需的LTQ-Orbitrap Velos添加到该资源中将大大减少样本积压，并提供5-10倍的灵敏度，使研究人员能够比目前可能的情况更深入地研究人类和其他复杂的蛋白质组。LTQ-Orbitrap Velos的速度越快，就能在SILAC或其他实验中从每种蛋白质中鉴定和量化更多的多肽，这将提供更积极的蛋白质鉴定，也将增加观察到的蛋白质表达差异的统计学意义。整合到LTQ-Orbitrap Velos中的先进技术也使其成为支持已成为该资源标志的磷酸蛋白质组学技术的理想平台。该应用程序得到了耶鲁大学的49个主要用户/研究人员和包括Mt.西奈医学院、洛克菲勒大学、芝加哥大学和斯坦福大学，他们建议开展51个依赖于所要求的仪器的研究项目。所需仪器的拟议主要用户包括四个与凯克实验室密切相关并具有强大蛋白质组学成分的NIH中心的成员(即耶鲁/NIDA神经蛋白质组中心、NCI资助的耶鲁癌症中心的蛋白质组学共享资源、NIAID资助的东北生物防御英才中心的蛋白质组学核心，以及即将进行竞争更新的耶鲁/NHLBI蛋白质组学中心)。这项提议的优势是多方面的，包括非常强大的机构支持；凯克实验室30年的成就记录，这是学术界最古老和最大的生物技术核心实验室之一；以及凯克实验室50名训练有素、非常敬业的工作人员。

DESCRIPTION (provided by applicant): Funding is requested towards the purchase of a Waters nanoACQUITY Ultra Performance LC and Thermo Fisher Scientific nano-electrospray ionization Linear Quadrupole Ion Trap - Orbitrap (LTQ-Orbitrap) Velos mass spectrometer system which would provide a uniquely high performance mass spectrometer for the MS & Proteomics Resource within the Keck Laboratory at Yale University. The LTQ-Orbitrap Velos is one of the newest Orbitrap MS platforms in its class that has sub-parts-per-million mass accuracy, a new atmospheric pressure interface lens design, and other high pressure ion trapping and ion transmission characteristics that together make the new Velos platform 2 times faster and 5-10 times more sensitive than its Model XL predecessor. Currently, Yale University has one LTQ-Orbitrap XL that is available on a service charge basis and that is located within the MS & Proteomics Resource of the Keck Laboratory. This Resource annually provides proteomics technologies to about 100 Yale and 110 non-Yale investigators at 90 institutions. The demand for technologies supported by its LTQ-Orbitrap XL has increased so rapidly that the delay for SILAC, Label-Free Quantitation, phosphoproteomics, and many other technologies dependent upon the LTQ-Orbitrap Velos has now reached >75 days with this instrument being run continuously at maximum capacity. Addition of the requested LTQ-Orbitrap Velos to this Resource would greatly reduce the sample backlog and provide a 5- 10 fold increase in sensitivity that would enable investigators to reach deeper into the human and other complex proteomes than is currently possible. The higher speed of the LTQ-Orbitrap Velos would enable more peptides to be identified and quantified from each protein in a SILAC or other experiment which would provide more positive protein identifications and also would increase the statistical significance of observed differences in protein expression. Advances incorporated into the LTQ-Orbitrap Velos also make it an ideal platform to support the phosphoproteomics technologies that have become a hallmark of this Resource. This application is supported by 49 major users/investigators at Yale and 8 other institutions including Mt. Sinai School of Medicine, Rockefeller U., the U. of Chicago, and Stanford U. who propose to carry out 51 research projects that depend upon the requested instrumentation. The proposed major users of the requested instrumentation include members of four NIH Centers that are closely associated with the Keck Laboratory and that have strong proteomics components (i.e. Yale/NIDA Neuroproteomics Center, the Proteomics Shared Resource in the NCI-funded Yale Cancer Center, the Proteomics Core of the NIAID-funded Northeast Center of Excellence for Biodefense, and the Yale/NHLBI Proteomics Center which has a pending competitive renewal). The strengths of this proposal are manifold and include very strong institutional support; the 30 year record of achievement of the Keck Laboratory, which is one of the oldest and largest biotechnology core labs in academia; and the 50 very highly trained and uncommonly dedicated staff that are the Keck Laboratory.