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

描述（由申请人提供）：本拨款申请要求从Applied Biosystems （AB）购买基质辅助激光解吸/电离飞行时间串联质谱仪（4800 Plus MALDI-TOF/TOF-MS），用于脂质组学和糖组学研究，这在不断发展的系统生物学科学中至关重要。这些研究领域目前正在成为诊断和新药物治疗发展的焦点，并了解传染病，癌症和代谢紊乱。AB 4800MALDI将配备以线性模式获取高分子量分析数据，以反射模式获取高分辨率分析数据，并在正离子和负离子模式下选择真前体MS/MS。该仪器将由俄亥俄州立大学（OSU）校园化学仪器中心的质谱和蛋白质组学设施（MS&P）进行管理。该仪器将填补我院脂质组学和糖组学研究缺失的重要空白。目前MS&P设施中可用的仪器能够进行高通量LC-MS/MS (LTQ)，分子式测定（Q-TOF和LCT），常规LC-MS (Q-TOF和LCT)，肽和聚合物筛选（Reflex MALDI）以及需要高分辨率LC-MS/MS的高级蛋白质组学（Orbitrap）。然而，没有一种仪器能够以足够高的灵敏度和分辨率从生物样品中获取数据，也没有能力进行碰撞诱导解离（CID）以进行MALDI结构测定。本提案的研究小组对与感染性和代谢性疾病相关的脂质、聚糖和碳水化合物的检查感兴趣。所要求的仪器满足了研究活动需要MALDI和CID功能的主要、次要和其他用户的需求。描述的主要研究活动包括：i)结核分枝杆菌（M.tb）和人类宿主内的其他分枝杆菌感染；ii)沙门氏菌和弗朗西斯菌在人类宿主体内生存的分子机制；iii)围绝经期妇女干眼病的临床研究；4)膳食脂肪酸与2型糖尿病关系的临床研究。描述的小项目包括结核分枝杆菌细胞包膜对肺微环境的适应，分析眼病不同状态下泪膜中发现的蛋白质上的聚糖部分，以及早产儿高频/高压通气后的高氧肺损伤。俄勒冈州立大学脂质科学小组的其他研究人员将使用所要求的仪器，包括营养学、皮肤、物理科学、牙科、验光、兽医学和癌症研究。除了作为我们研究项目成功的重要工具外，ab4800maldi还将加强与外部用户的合作。

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.