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

点击翻译按钮获取中文摘要

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.