Orbitrap mass spectrometer for biomedical research

描述（由申请人提供）：为了提高芝加哥地区研究人员的NIH资助的蛋白质组学研究的生产率，我们建议将现有的离子阱质谱仪（Thermo LCQ Deca）换成Thermo LTQ Orbitrap Velos高分辨率质谱仪。该仪器将位于芝加哥生物医学联盟/伊利诺伊大学芝加哥分校研究资源中心蛋白质组学和信息学服务设施，该设施为西北大学、芝加哥大学、阿贡国家实验室和伊利诺伊大学芝加哥分校的研究人员提供服务。升级系统具有利用现有Dionex 2D微毛细管HPLC系统和专门为质谱设计的实验室空间的经济优势。我们应用的一个独特方面是，我们设计了一个Agilent Chip Cube微毛细管LC-纳米电喷雾系统，用于在LTQ Orbitrap Velos上运行。我们已经通过在加州的Thermo演示实验室的LTQ轨道阱Velos上操作我们改进的Chip Cube来证明这种能力。Chip Cube可在LTQ Orbitrap Velos上实现更快、更可重复、更可靠的微毛细管HPLC分离和纳米电喷雾。据我们所知，没有其他实验室具有这种能力。我们的设施是在芝加哥大都市区的蛋白质组学活动的中心，并由芝加哥生物医学联盟资助，以促进生物医学研究人员之间的校际合作研究。在2009年期间，我们的质谱实验室进行了近11，000小时的质谱分析（不包括维护时间），并在线性离子阱质谱仪和我们的FTMS上使用LC-MS/MS进行了近4，000小时的蛋白质组学研究。我们拥有245名蛋白质组学研究人员的用户群。拟议的LTQ Orbitrap Velos质谱仪是一种高分辨率、高质量准确度的仪器，将提高基于LC-MS/MS的蛋白质组学的效率和生产力，例如蛋白质鉴定和涉及稳定同位素标记的蛋白质和肽的应用。一些将受益的应用包括使用iTRAQ、ICAT和SILAC对蛋白质表达进行定量研究，以及对蛋白质结构和功能进行H/D交换测量。由于FTMS的扫描速度慢或灵敏度限制，或者离子阱在低质量和低分辨率下的灵敏度差，这些测量目前在我们的设施中不实用。Chip Cube技术的使用将通过减少运行时间和提高纳米电喷雾可靠性来提高通量，而Orbitrap Velos的高分辨率将提高信息学通量。拥有5个博士学位。我们的设施拥有质谱、蛋白质组学和信息学方面的高级专家，是蛋白质组学研究和教育的主要区域资源，拟议的质谱仪将提高我们所有NIH资助的研究人员的研究生产力。

DESCRIPTION (provided by applicant): To enhance the productivity of NIH-funded proteomics research of Chicago-area researchers, we propose to trade in an existing ion trap mass spectrometer (Thermo LCQ Deca) for a Thermo LTQ Orbitrap Velos high resolution mass spectrometer. This instrument will be located in the Chicago Biomedical Consortium/ University of Illinois at Chicago Research Resource Center Proteomics and Informatics Services Facility which serves researchers at Northwestern University, the University of Chicago, Argonne National Laboratory, and the University of Illinois at Chicago. Upgrading the system has the economic advantage of utilizing an existing Dionex 2D microcapillary HPLC system and laboratory space designed specifically for mass spectrometry. A unique aspect of our application is that we have engineered an Agilent Chip Cube microcapillary LC-nanoelectrospray system to operate on the LTQ Orbitrap Velos. We have demonstrated this capability by operating our modified Chip Cube on an LTQ Orbitrap Velos at the Thermo demonstration laboratory in California. The Chip Cube enables faster and more reproducible and reliable microcapillary HPLC separations and nanoelectrospray on the LTQ Orbitrap Velos than would otherwise be possible. To the best of our knowledge, no other laboratory has this capability. Our facility is the center of proteomics activity in the Chicago metropolitan area and is subsidized by the Chicago Biomedical Consortium to facilitate intercampus collaborative research between biomedical investigators. During 2009, our mass spectrometry laboratory carried out almost 11,000 hours of mass spectrometric analyses (not counting maintenance time), and almost 4,000 hours were devoted to proteomics using LC- MS/MS on a linear ion trap mass spectrometer and our FTMS. We have a user base of 245 proteomics investigators. The proposed LTQ Orbitrap Velos mass spectrometer is a high resolving power, high mass accuracy instrument that will enhance the efficiency and productivity of LC-MS/MS based proteomics such as protein identification and applications involving stable isotope labeled protein and peptides. Some of the applications that will benefit include quantitative studies of protein expression using iTRAQ, ICAT and SILAC and H/D exchange measurements of protein structure and function. These measurements are not currently practical in our facility due to slow scan speeds or sensitivity limitations on the FTMS or poor sensitivity at low mass and low resolution on the ion trap. The use of the Chip Cube technology will enhance throughput by reducing run times and increasing nanolectrospray reliability, and the high resolution of the Orbitrap Velos will enhance informatics throughput. Staffed with five Ph.D. level experts in mass spectrometry, proteomics and informatics, our facility is a major regional resource for proteomics research and education, and the proposed mass spectrometer will enhance the research productivity of all of our NIH-funded investigators.