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

配备 ESI 和 Dionex Ultimate 3000 HPLC 系统的 Thermo Electron LTQ Orbitrap XL ETD

• 批准号: 7838746
• 负责人: MARK R CHANCE
• 金额: $ 98.3万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-08 至 2012-07-07
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7838746
• 关键词: Animals; Award; Bioinformatics; Biological Markers; Bone Marrow Transplantation; Cancer Biology; Cancer Center; Cell Line; Cells; Center for Translational Science Activities; Chromatography; Contract Services; Cytokine Signaling; Detection; Development; Diabetes Mellitus; Disease; Disease remission; Electrons; Fees; Foundations; Funding; Goals; Graft Rejection; Grant; Health; High Pressure Liquid Chromatography; Human Resources; Immunity; Insulin; Knowledge; Label; Liquid substance; Maintenance; Mass Spectrum Analysis; Microbial Biofilms; Minor; Modification; Occupations; Pathogenesis; Patients; Peer Review; Phenotype; Plasma; Post-Translational Protein Processing; Preventive; Prions; Proteins; Proteomics; Publications; Research; Resolution; Running; Sampling; Schedule; Science; Signal Pathway; Signal Transduction; Solutions; Stem cells; Structure; Support Groups; System; Techniques; Technology; Time; Tissues; Training; United States National Institutes of Health; Universities; Yeasts; base; experience; immune activation; instrument; novel; programs; protein expression; protein structure; public health relevance; skin disorder

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.

描述（由申请人提供）：该提案将通过从Dionex购买Thermo-Scientific LTQ Orbitrap XL（带ETD）和先进的色谱系统，提高凯斯西储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。该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增加、资助赠款增加、出版物增加以及每月运行的样本增加。 特别是，与需要高分辨率、高灵敏度和高质量准确度质谱分析的技术直接相关的用户需求和蛋白质组学赠款目前增长最快。提案中概述的主要和次要用户的蛋白质组学研究得到了四十多个R型，P型（或U型）赠款以及基金会和试点赠款的支持。此外，该中心还运营着七个由NIH资助的主要中心的同行评审核心，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，目前的两个高分辨率仪器，一个Orbitrap和一个LTQ-FT，都来自Thermo，被当前的项目超额预订，而最近获得的一些NIH赠款，包括几个依赖于高分辨率仪器的计划项目和中心赠款，表明当前的积压将在未来12-24个月内大幅增加。特别是，无标记蛋白质组学研究，以确定蛋白质丰度的变化，识别翻译后修饰，以了解细胞信号传导，和结构质谱研究，以了解构象变化和蛋白质的结构在溶液中，是三个最快速增长的蛋白质组学计划在该中心。这些程序将显著受益于所要求的Orbitrap仪器的高灵敏度和高准确度，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涵盖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，艾滋病毒-艾滋病发病机制，免疫激活生物标志物，糖尿病及其并发症，胰岛素结构-功能，免疫和皮肤病，骨髓移植排斥，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。 公共卫生相关性：关于蛋白质表达、结构、相互作用和修饰的变化的知识为我们提供了关于发育和疾病背后的信号通路变化的关键信息，并获得了组织、细胞或细胞系以及从患者或动物非侵入性收集的血浆或其他液体的表型的新表征。这些信息为我们提供了对疾病和健康的病理生理学基础的理解，而蛋白质中这种变化的可靠和可重复的检测可以用作疾病存在及其缓解的生物标志物。这些生物医学科学研究通过以多种方式提供和保留工作来支持ARRA的目标。LTQ Orbitrap XL ETD和Ultimate 3000 HPLC系统的制造和安装将创造或保留至少5个工作岗位。制造Orbitrap将需要3-4个月的人工时间。构建Dionex仪器将需要2个月的人工时间。此外，Thermo Electron和Dionex的人员安装、熟悉和培训Orbitrap和Ultimate 3000的使用将需要三周时间。还需要四名项目经理，两名监督制造，两名监督安装和培训。该提案还包括一份为期3年的轨道阱服务合同，该合同将需要每年进行预防性维护。此外，我们将安排Ultimate 3000的预防性维护。