Thermo Electron LTQ Orbitrap XL ETD with ESI and Dionex Ultimate 3000 HPLC System

描述（由申请人提供）：该提案将通过购买热科学LTQ Orbitrap XL（带ETD）和来自Dionex的先进色谱系统来增强Case Western Reserve大学蛋白质组学和生物信息学中心（CPB）的质谱分析能力和研究通量。本中心支持的用户群体正在迅速增长，这反映在收取的用户费用增加、资助赠款增加、出版物增加和每月运行的样品增加上。特别是，用户需求和蛋白质组学补助金直接与需要高分辨率、高灵敏度和高质量精度质谱法的技术相关，目前增长最快。提案中概述的主要和次要用户的蛋白质组学研究得到了40多个r型，p型（或u型）赠款以及基金会和试点赠款的支持。此外，该中心还运营着美国国立卫生研究院资助的主要中心的七个同行评审核心，包括转化科学合作中心的转化技术核心和癌症中心蛋白质组学核心。不幸的是，目前的两种高分辨率仪器，Orbitrap和LTQ-FT，都来自Thermo，目前的项目已经超额预定，而最近NIH的一些拨款，包括一些依赖高分辨率仪器的项目项目和中心拨款，表明目前的积压将在未来12-24个月内大幅增加。特别是，用于鉴定蛋白质丰度变化的无标记蛋白质组学研究，用于了解细胞信号的翻译后修饰鉴定，以及用于了解溶液中蛋白质构象变化和结构的结构质谱研究，是该中心发展最快的三个蛋白质组学项目。这些项目将显著受益于Orbitrap仪器的高灵敏度和高精度，而ETD能力将增强蛋白质鉴定和蛋白质修饰的鉴定。本提案中确定的用户的研究跨越了广泛的生物医学科学和许多疾病，包括：干细胞，癌症生物学，HIV-AIDS发病机制，免疫激活生物标志物，糖尿病及其并发症，胰岛素结构-功能，免疫和皮肤疾病，骨髓移植排斥反应，酵母生物膜形成，朊病毒结构和功能，细胞因子信号传导等。他们在高分辨率和高精度仪器方面拥有丰富的经验，并在蛋白质组学方面发表了许多文章。

DESCRIPTION (provided by applicant): This proposal will enhance mass spectrometry capability and research throughput of the Center for Proteomics and Bioinformatics (CPB) at Case Western Reserve University by the purchase of a Thermo-Scientific LTQ Orbitrap XL (with ETD) and advanced chromatography systems from Dionex. The user group supported by this Center is growing rapidly, as reflected in increases in user fees collected, increases in funded grants, publications, and increases in samples run per month. In particular, user needs and proteomics grants directly tied to techniques that require high-resolution, high-sensitivity, and high mass accuracy mass spectrometry are currently increasing the fastest. The major and minor users' proteomics research as outlined in the proposal is supported by over forty R-type, P-type (or U-type) grants plus foundation and pilot grant support. Also, the Center operates seven peer-reviewed Cores of major NIH funded Centers, including the Translational Technologies Core of the Center for Translational Science Collaborative and the Cancer Center Proteomics Core. Unfortunately, the two current high-resolution instruments, an Orbitrap and an LTQ-FT, both from Thermo, are overbooked with current projects while the very recent award of a number of NIH grants, including several program project and center grants dependent on high-resolution instruments, suggests that the current backlog will increase substantially over the next 12-24 months. In particular, label-free proteomics studies to identify protein abundance changes, identification of post-translational modifications to understand cell signaling, and structural mass spectrometry studies to understand conformational changes and structure of proteins in solution, are the three most rapidly growing proteomics programs in the Center. These programs will significantly benefit from the high sensitivity and high accuracy of the requested Orbitrap instrument while the ETD capability will enhance protein identification and identification of protein modifications. The research of the users identified in this proposal spans a wide range of biomedical science and many diseases including: stem cells, cancer biology, HIV-AIDS pathogenesis, immune activation biomarkers, diabetes and its complications, insulin structure-function, immunity and skin disease, bone marrow transplant rejection, yeast biofilm formation, prion structure and function, cytokine signaling, and many others. They have extensive experience with high-resolution and high accuracy instruments and many publications in proteomics. PUBLIC HEALTH RELEVANCE: Knowledge about changes in protein expression, structure, interactions and modifications gives us a crucial information on the changes in signaling pathways underlying development and disease and procuring a novel characterization of phenotype for tissues, cells or cell lines, as well as plasma or other fluids collected non-invasively from patients or animals. This information provides us with understanding of the pathophysiological basis of disease and health while the reliable and reproducible detection of such changes in proteins can be utilized as biomarkers for the presence of disease and its remission. These biomedical science studies support the ARRA goals by providing and retaining jobs in a number of ways. Manufacturing and installation of the LTQ Orbitrap XL ETD and the Ultimate 3000 HPLC System will create or retain at least 5 jobs. Manufacturing the Orbitrap will require between 3-4 months personnel time. Building the Dionex instrument will require 2 months personnel time. Additionally, installation, familiarization, and training on utilization of the Orbitrap and Ultimate 3000 by personnel from Thermo Electron and Dionex will require three weeks. Four project managers are also required, two to oversee manufacturing and two to oversee the installation and training. The proposal also includes a 3-year service contract for the Orbitrap which will require yearly preventive maintenance. Additionally, we will schedule preventative maintenance of the Ultimate 3000.