Acquisition of a MALDI-TOF/TOF MS for Glycomic and Lipidomic Research

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DESCRIPTION (provided by applicant): This grant application is requesting funding to purchase a Matrix-Assisted Laser Desorption/Ionization Time-of- Flight tandem mass spectrometer (4800 Plus MALDI-TOF/TOF-MS) from Applied Biosystems (AB) for lipidomic and glycomic research, critical in evolving systems biology science. These research fields are currently emerging as a focus of the development of diagnostics and novel drug therapies, and understanding infectious diseases, cancer, and metabolic disorders. The AB 4800MALDI will come equipped to acquire data in linear mode for high molecular weight analysis, reflectron mode for high resolution analysis and true precursor selected MS/MS in both positive and negative ion mode. The instrument will be located and administered by the Mass Spectrometry and Proteomics Facility (MS&P) in the Campus Chemical Instrument Center at The Ohio State University (OSU). This instrument will complete an important missing niche for lipidomic and glycomic research at our Institution. The current instrumentation available in the MS&P Facility is capable of high-throughput LC-MS/MS (LTQ), molecular formula determination (Q-TOF and LCT), routine LC- MS (Q-TOF and LCT), peptide and polymer screening (Reflex MALDI) and advanced proteomics requiring high resolution LC-MS/MS (Orbitrap). However, none of the instruments has the ability to acquire data from biological samples with high enough sensitivity and resolution, nor do they have the ability to perform collision- induced dissociation (CID) for structural determination by MALDI. The team of investigators in this proposal is interested in the examination of lipids, glycans, and carbohydrates related to infectious and metabolic diseases. The requested instrumentation addresses the needs of major, minor and other users whose research activities require MALDI and CID capabilities. The major research activities described involve: i) Mycobacterium tuberculosis (M.tb) and other Mycobacterium spp infections within the human host; ii) Molecular mechanisms used by Salmonella and Francisella spp to survive within the human host; iii) Clinical research on dry eye disease in perimenopausal women; and iv) Clinical investigation of the relationship of dietary fatty acids with type II diabetes. Minor projects described are adaptation of the M.tb cell envelope to the lung microenvironment, analysis of glycan moieties on proteins found in the tear film in various states of eye disease, and hyperoxic lung injury following high frequency/pressure ventilation in premature infants. Other investigators within the established OSU Lipid Science Group that will use the requested instrument involve nutrition, skin, physical sciences, dentistry, optometry, veterinary medicine and cancer research. In addition to being an essential tool for the success of our research programs, the requested AB 4800MALDI will enhance collaborations with external users. PUBLIC HEALTH RELEVANCE: The fields of lipidomics and glycomics are now recognized as playing a critical role in systems biology and are now becoming the focus of the development of diagnostic tests, novel drug therapies, and understanding infectious diseases, cancer and metabolic disorders. Given the importance of lipidomics and glycomics in these fields, the team of investigators in this proposal is interested in the examination of lipids, glycans and carbohydrates in infectious and metabolic diseases. Matrix-Assisted Laser Desorption/Ionization Time-of-Flight mass spectrometry (MALDI-TOF-MS) analysis of lipids and glycans is sensitive, less affected by impurities than electrospray, involves relatively easy sample preparation and is an excellent analytical technique for rapid screening of lipids and glycans from biological matrices. The ability to perform true precursor ion selected collision-induced dissociation (CID) on these samples will also allow for fine structure elucidation of complex mixtures.