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

点击翻译按钮获取中文摘要

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.