Acquisition of a Hybrid Quadrupole, Ion Mobility Separation, Orthogonal Accelerat

描述（由申请人提供）：本申请请求支持从沃茨公司购买混合四极杆、离子迁移率分离、正交加速飞行时间串联质谱仪（Synapt HDMS），该质谱仪与完全集成的超高效液相色谱（nanoAcquityUPLC）平台连接。这种最先进的仪器将被用作康奈尔大学蛋白质组学和质谱（MS）设施的共享研究资源，并将对28个NIH资助的赠款至关重要。康奈尔大学对蛋白质组学和MS的兴趣越来越大，过去3年来该机构处理的样品数量呈指数级增长就证明了这一点（图2）。拟议的仪器将补充现有的仪器在康奈尔大学，特别是在其应用程序分析完整的蛋白质复合物，蛋白质-蛋白质/配体相互作用和探测蛋白质构象。这种高度灵敏和准确的质谱仪将提供蛋白质结构域作图和肽作图的能力，从而可靠地鉴定蛋白质修饰和更好地对没有可用基因组序列的蛋白质进行从头测序。这种高分辨率仪器对于使用稳定同位素标记（SILAC和iTRAQ）的定量蛋白质组学研究的成功至关重要。UPLC分离功能与MSE扫描功能相结合，提高了仪器的动态范围，增强了低丰度蛋白质的鉴定，并实现了无标记比较蛋白质组学。此外，该仪器包括离子迁移率和时间对齐平行（TAP）碎片化功能，这将提供更高的特异性和更可靠的蛋白质和其他生物分子的结构表征。所提出的仪器的特殊优势是：i）离子迁移率分离，其提供了一种除了质量之外还估计分子大小、形状和电荷的手段; ii）TAP片段化用于结构表征; iii）高质量准确度（通过LockSpray < 2 ppm RMS）、高分辨率（17，500）和宽动态范围（4阶线性度）对于提高数据质量至关重要; iv）高占空比和快速TDC（4GHz），用于快速数据采集速率; v）宽m/z范围（100，000 m/z），用于能够分析> 1，000 kDa蛋白质复合物的TOF分析仪; vi）具有MSE模式的UPLC-MS/MS，用于并行数据采集，使得能够灵敏地分析具有宽动态浓度的分析物的复杂样品。拟议的仪器将加强核心设施中现有的电喷雾质谱技术。该仪器的收购将对超过20个NIH资助的研究项目产生巨大影响，并将改善康奈尔大学的研究基础设施以及学生和研究人员的教育/培训计划。它将加速和促进康奈尔大学教师在六个与生命科学研究相关的学院的广泛研究项目，并有助于保持康奈尔大学的竞争地位。公共卫生相关性：这种应用对公共卫生的相关性是通过28个国家卫生研究院支持的研究项目建立的，这些项目将从这种仪器的购买中受益。本文所要求的仪器拥有的技术能力远远超过这些研究人员目前拥有的能力，因此将使他们能够通过收集更多更好的数据来实现这些资助项目的目标。提高与这些项目有关的数据的质量和数量对于实现公共卫生目标至关重要，每个项目都获得了优先资助。

DESCRIPTION (provided by applicant): This application requests support for the acquisition of a hybrid Quadrupole, Ion Mobility separation, orthogonal acceleration Time-of-Flight Tandem Mass Spectrometer (Synapt HDMS), interfaced with a fully integrated ultra-high performance liquid chromatography (nanoAcquityUPLC) platform from Waters Corporation. This state-of-the-art instrument will be utilized as a shared research resource in the Cornell Proteomics and Mass Spectrometry (MS) Facility and will be essential to 28 active NIH supported grants. There is a growing interest in proteomics and MS at Cornell, as is evidenced by the exponential growth in the number of samples processed by the facility over the past 3 years (Figure 2). The proposed instrument will complement the existing instrumentation in Cornell, particularly with respect to its applications for analysis of intact protein complexes, protein-protein/ligand interactions and probing protein conformation. This highly sensitive and accurate mass instrument will provide capabilities in protein domain mapping and peptide mapping resulting in confident identification of protein modifications and better de-novo sequencing of proteins for which there is no available genome sequence. This high resolution instrument will be critical to the success of quantitative proteomics studies using stable isotope labeling (SILAC and iTRAQ). The power of UPLC separation combining with MSE scan feature improves the dynamic range of the instrument enhancing the identification of low abundance proteins and enables label-free comparative proteomics. Furthermore, the instrument includes both ion mobility and Time Aligned Parallel (TAP) fragmentation features which will provide increased specificity and more confident structural characterization of proteins and other bio-molecules. The particular strengths of the proposed instrument are: i) Ion Mobility separation which provides a means to estimate molecular size, shape and charge in addition to mass; ii) TAP fragmentation for structural characterization; iii) High mass accuracy (< 2 ppm RMS via the LockSpray), high resolution (17,500) and wide dynamic range (4 orders of linearity) critical for improved data quality; iv) High duty cycle and fast TDC (4GHz) for fast data acquisition rates; v) Wide m/z range (100,000 m/z) for TOF analyzer capable of analysis > 1,000 kDa protein complexes; vi) UPLC-MS/MS with MSE mode for a parallel data acquisition enabling sensitive analysis of complex samples with a wide dynamic concentration of analytes. The proposed instrument will enhance the electrospray MS technologies available in the core facility. The acquisition of this instrument will have an enormous impact on more than two dozens of NIH-funded research projects, and will improve the research infrastructure of Cornell University and the education/training programs of students and researchers. It will accelerate and facilitate a wide range of research projects of Cornell faculty across six colleges relevant to life sciences research, and help to maintain Cornell's competitive position. PUBLIC HEALTH RELEVANCE: The relevance of this application to public health is established through the 28 odd NIH supported research programs that would benefit from the acquisition of this instrumentation. The instrumentation requested herein possesses technical abilities that far exceed those that are currently available to these researchers and thus would allow them to accomplish the goals of these funded projects through the gathering of more and better data. The improved quality and quantity of data gathered in connection with these projects is critical to the achievement of the public health goals for which each was granted priority funding.