Orbitrap mass spectrometer for biomedical research

描述(申请人提供)：为了提高芝加哥地区研究人员在美国国立卫生研究院资助的蛋白质组学研究的效率，我们建议将现有的离子陷阱质谱仪(Thermo LTQ Deca)换成Thermo LTQ Orbitrap Velos高分辨率质谱仪。该仪器将设在芝加哥生物医学联盟/伊利诺伊大学芝加哥研究资源中心蛋白质组学和信息学服务设施，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。升级该系统具有经济优势，可以利用现有的Dionex 2D微毛细管高效液相色谱系统和专门为质谱学设计的实验室空间。我们应用的一个独特方面是，我们设计了一种安捷伦芯片立方体微毛细管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Thermo演示实验室的LTQ Orbitrap Velos上操作我们的改装芯片立方体来展示这种能力。芯片立方体使LTQ Orbitrap Velos上的微毛细管高效液相色谱分离和纳米电喷雾比其他任何方式都更快、更可靠和更可靠。据我们所知，没有其他实验室拥有这种能力。我们的设施是芝加哥大都市区蛋白质组学活动的中心，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。2009年，我们的质谱仪实验室进行了近11,000小时的质谱分析(不包括维护时间)，并使用线性离子陷阱质谱仪和我们的FTMS上的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基础。建议的LTQ Orbitrap Velos质谱仪是一种高分辨率、高质量精度的仪器，将提高基于LC-MS/MS的蛋白质组学的效率和生产率，例如蛋白质鉴定和涉及稳定同位素标记蛋白质和多肽的应用。一些有益的应用包括使用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。这些测量目前在我们的设备中是不实用的，因为扫描速度慢，或者FTMS的灵敏度限制，或者离子陷阱在低质量和低分辨率下的低灵敏度。芯片立方体技术的使用将通过减少运行时间和提高纳米喷雾的可靠性来提高吞吐量，而Orbitrap Velos的高分辨率将提高信息学吞吐量。我们的设施拥有五名质谱学、蛋白质组学和信息学方面的博士水平的专家，是蛋白质组研究和教育的主要地区性资源，拟议的质谱仪将提高我们所有NIH资助的研究人员的研究效率。

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.