Acquisition of a Matrix-Assisted Laser Desorption Time of Flight (MALD-TOF) Mass Spectrometer

本提案中要求的仪器是布鲁克BIFLEX基质辅助激光解吸飞行时间质谱仪（MALD-TOF-MS），配有HIMAS检测器、FAST附件、SCOUT多样品膜靶和观察光学器件、Nd：YAG激光器和PolyMASS软件，用于Mn、Mw和多分散指数测量，并可轻松与GPC数据进行比较。布鲁克BIFLEX专为生物分析和聚合物科学研究而设计。分子离子和结构显著碎片离子的外部质量校准通常为0.01%至0.1%。BIFLEX非常灵敏，可提供亚皮摩尔样品的分子量信息。HIMAS检测器在代替EM检测器使用时，提供了更高的高质量灵敏度，并且被制造商推荐用于高质量分子的线性分析，特别是聚合物。该检测器的动态范围为20，000至150，000，可提供最大的质量精度。 质谱仪的使用将由生物分子科学研究所（IBS）监督，仪器的日常使用、维护和调度由南佛罗里达大学新生物科学设施的研究助理教授和研究生研究助理监督。IBS是一个由来自坦帕湾地区四所USF学院和研究机构的78名科学家组成的联盟，旨在促进跨越生物学、化学和分子生物学界限的研究互动，并提供一种合作机制。此外，Rebecca O 'Malley博士将担任本提案中所有项目的顾问。她拥有博士学位。在质谱和经营一个积极的质谱研究计划，自1980年以来一直存在于USF。她还建立了多光子电离飞行时间质谱的研究计划，最近又建立了激光解吸飞行时间质谱的研究计划。 使用MALD-TOF-MS的研究项目将集中在聚合物，肽和蛋白质的表征上。Newkome博士（PI）、O 'Malley博士、Harmon博士和Garcia-Rubio博士专注于聚合物。他们的项目涉及新型树枝状聚合物的表征-影响阳离子附着到合成聚合物的因素的调查;耐辐射聚合物增透剂和光学聚合物，优化闪烁体和光纤包层聚合物的耐光辐射性;以及复杂聚合物分子的表征，如共聚物和支化聚合物。Solomonson、Barber、Neame和Ferreira博士专注于蛋白质和肽表征领域。 典型的实验包括二硫键排列的分析，翻译后修饰位点的分析，以及辅基连接的构象。 所要求的仪器具有广泛的质量范围，即小肽和聚合物，以及完整的蛋白质和较大的聚合物。 它在低质量水平下的极端准确性对于合成化学家和定义肽片段化模式的研究人员至关重要。这些特点使该仪器特别适合在该机构进行的研究活动。此外，据我们所知，在USF或坦帕湾地区没有质谱仪。这是一个严重的问题，教育和研究在这个大（35,000名学生）研究II大学。拥有MALD-TOF质谱仪将允许将“动手”经验整合到既定的研究生课程中，增加学生对MS应用于化学和生物化学问题的理解。此外，该仪器将可用于论文和论文研究，从而提高研究生调查的质量。 当IBS教师对USF最需要的仪器进行民意调查，以加强正在进行的研究并吸引优秀的年轻教师和学生时，他们压倒性地投票支持可以处理大型聚合物和肽的质谱仪。对于一些蛋白质和聚合物化学家来说，这一需求是他们研究计划的核心，而对于其他人来说，这将使他们能够扩展到新的方向。质谱法用于大型合成聚合物和生物分子是相对较新的，但却是一个不断发展和令人兴奋的领域。合作，多样化和互动的科学家，大型研究生和研究项目的结合以及生物分子科学研究所的参与应该确保这种仪器在最大程度上用于改善USF的生物分子科学计划。

The instrument requested in this proposal is the Bruker BIFLEX Matrix-Assisted LASER Desorption Time-of Flight Mass Spectrometer (MALD-TOF-MS) with HIMAS Detector, FAST accessory, SCOUT multisample membrane target and observation optics, an Nd:YAG laser and PolyMASS software for Mn, Mw and polydispersity index measurements with easy comparison to GPC data. The Bruker BIFLEX is specifically designed for research in bio-analytical and polymer sciences. External mass calibration for both molecular ions and structurally significant fragment ions is typically 0.01% to 0.1%. The BIFLEX is extremely sensitive and provides molecular weight information for sub picomoles of a sample. The HIMAS detector, when used in place of an EM detector, provides increased high-mass sensitivity and is recommended by the manufacturer for the linear analysis of high mass molecules, particularly polymers. This detector has a dynamic range of 20,000 to 150,000, providing maximum mass accuracy. The use of the mass spectrometer will be overseen by the Institute for Biomolecular Science (IBS) with the daily use, maintenance and scheduling of the instrument supervised by a Research Assistant Professor and graduate research assistant in the new Bioscience Facility at University of South Florida. IBS is a consortium of 78 scientists from four USF colleges and research institutes in the Tampa Bay area that promotes research interactions that cross the boundaries between biology, chemistry and molecular biology and offers a mechanism for collaboration. In addition, Dr. Rebecca O'Malley will act as a consultant on all the projects included in this proposal. She holds a Ph.D. in mass spectrometry and operates an active mass spectrometry research program that has been in existence since 1980 at the USF. She also established a research program in multiphoton ionization time-of-flight mass spectrometry and, more recently, one in laser desorption time-of-flight mass spectrometry. Research projects using the MALD-TOF-MS will center on the characterization of polymers, peptides and proteins. Dr. Newkome (PI), Drs. O'Malley, Harmon, and Garcia-Rubio, concentrate their efforts on polymers. Their projects involve the characterization of novel dendrimeric polymers- the investigation of the factors that affect cation attachment to synthetic polymers; radiation resistant polymer scintillators and optical polymers that optimize optical radiation resistance in scintillator and fiber cladding polymers; and the characterization of complex polymer molecules such as copolymers and branched polymers. Drs. Solomonson, Barber, Neame and Ferreira focus their efforts in the area of protein and peptide characterization. Typical experiments include the analysis of disulfide bond arrangements, analysis of sites of post-translational modification, and the conformation of attachment of prosthetic groups. The requested instrumentation has a broad mass range that is small peptides and polymers, as well as intact proteins and larger polymers. Its extreme accuracy at low mass levels is critical for synthetic chemists and investigators who are defining peptide fragmentation patterns. These features make this instrument uniquely suited to the research activities conducted at this institution. Furthermore, to the best of our knowledge, there is no mass spectrometer at USF or in the Tampa Bay area. This is a serious problem for education and research at this large (35,000 students) Research II university. Having a MALD-TOF Mass Spectrometer will allow the integration of "hands-on" experience into established graduate courses, increasing students' understanding of the applications of MS to chemical and biochemical problems. In addition, the instrument will be available for thesis and dissertation research and thus improve the quality of the graduate student investigations. When IBS faculty were polled about the instrument most needed at USF to augment ongoing research and att ract excellent young faculty and students, they overwhelming voted for a mass spectrometer that could handle large polymers and peptides. For some of the protein and polymer chemists the need is central to their research programs and for others it will allow them to expand into new directions. The use of mass spectrometry for large synthetic polymers and biomolecules is relatively recent but is a growing and exciting field. The combination of cooperative, diverse and interactive scientists, large graduate and research programs and the participation of the Institute for Biomolecular Science should ensure that this instrumentation is used to its fullest extent to improve the biomolecular science program at USF.