Acquisition of a Hybrid Quadrupole Time of Flight LC-MS/MS System for the Cornell

描述(由申请人提供)：新的质谱学(MS)技术为具有突破性发现潜力的创造性应用提供了一个特殊的机会。我们申请资助一台高性能Q-TOF质谱仪(AB SCIEX TripleTOF“5600)，与超高性能液相色谱系统(基于芯片的Ekpert”NanoLC 425)连接。这个最先进的系统将设在康奈尔大学生物技术资源中心(BRC)的蛋白质组学和质谱学设施中。将该仪器作为大学范围内共享的研究资源在BRC中实施，将使NIH资助的广泛研究能够以成本效益的方式获得这项至关重要的技术。康奈尔大学对蛋白质组学和质谱学服务的需求日益增长，这一点从该设施过去九年来处理的设施用户和样本数量的快速和持续增长中可见一斑。拟议的综合仪器系统将扩大设施的容量和能力，但不会重复核心设施的现有仪器，目前核心设施已接近满负荷使用。对于涉及复杂样品分析的应用而言，提高性能和容量的需求尤其关键，这些应用需要更高的扫描速度、高灵敏度和分辨率以及高精度的质量测量。所要求的集成系统将提供改进的动态范围，并将使从低丰度蛋白质和复杂样品中的翻译后修饰(PTM)中确定多肽成为可能。TripleTOF 5600将显著增强该设施的能力，以支持定性探索中的蛋白质组学发现研究，以及涉及复杂生物样本的高通量图谱和高分辨率量化研究。采用cHiPLC(R)系统的Ekpert“NanoLC 425是一种基于芯片的集成超高效液相，提供广泛的2D LC灵活性和连续的直接纳米流。它在使用基于芯片的柱的工作流选择方面具有灵活性，并为全局发现和更高吞吐量目标定量提供了广泛的流速范围。NanoLC 425系统将在MS之前提供自动样品处理和高分辨率多肽分离，这将使非常复杂的样品能够全天候连续分析。BRC在运行LC MS仪器系统和提供MS应用程序作为核心设施服务方面拥有丰富的经验。拟议的仪器系统将很容易地整合到该设施已建立的蛋白质组学工作流程中，并将大大扩展该设施的定量蛋白质组学管道。将仪器系统放置在BRC中将使NIH资助的不同群体的调查人员能够开展原本在经济上或技术上不可行的研究项目。这份提案描述了来自NIH资助的16个研究小组的39个项目，这些项目将利用拟议的系统。所要求的文书将使目前和未来由美国国立卫生研究院资助的各种项目受益。

DESCRIPTION (provided by applicant): New mass spectrometry (MS) technologies present an exceptional opportunity for creative applications with potential for breakthrough discoveries. We request funding a high performance Q-TOF mass spectrometer (AB SCIEX TripleTOF" 5600+), interfaced with an ultra-high performance liquid chromatography system (chip-based ekspert" nanoLC 425). This state-of-the-art system will be located in the Proteomics and Mass Spectrometry Facility of the Cornell University Biotechnology Resource Center (BRC). Implementation of the instrument in the BRC as a university-wide shared research resource will enable cost effective access to this vital technology for a broad range of NIH-funded research. There is a growing demand for proteomics and mass spectrometry services at Cornell, as shown by the rapid and continuing growth in the number of facility users and samples processed by the facility over the past nine years. The proposed integrated instrument system will expand the capacity and capabilities of the facility but not duplicate the existing instrumentation at the core facility, which is currently utilized at near full capacity. The need fr increased performance and capacity is particularly critical with respect to applications involving analysis of complex samples which require enhanced scan speed, high sensitivity and resolution, and highly accurate mass measurements. The integrated system requested will provide improved dynamic range and will enable confident identification of peptides from low abundance proteins and of post-translational modifications (PTMs) in complex samples. The TripleTOF 5600+ will significantly enhance the facility's capability to support proteomics discovery studies in qualitative exploration as well as high throughput profiling and high-resolution quantification studies involving complex biological samples. The ekspert" nanoLC 425 with the cHiPLC(R) system is an integrated chip-based UHPLC that provides broad 2D LC flexibility and continuous direct nano flow. It has flexibility in workflow selection using chip-baed columns and offers a broad range of flow rates for both global discovery and higher throughput targeted quantitation. The nanoLC 425 system will provide automated sample handing and high resolution peptide separation prior to MS, which will enable continuous 24/7 analysis of very complex samples. The BRC has extensive experience in running LC MS instrument systems and providing MS applications as core facility services. The proposed instrument system will be readily integrated into the facility's established proteomics workflows and will significantly expand the facility's quantitative proteomics pipeline. Placement of the instrument system in the BRC will enable a diverse group of NIH funded investigators to undertake research projects that would not otherwise be economically or technically feasible. This proposal describes 39 projects from 16 NIH funded investigator groups that will utilize the proposed system. The requested instrument will benefit a wide range of current and future NIH-funded projects.