Teacher
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TIMPERIO Anna Maria
(syllabus)
PRINCIPLES OF SCIENCES "OMIC" Introduction: • "omics" sciences and biology of complex systems. • The hypothesis revolution in retrospect and the search for new hypotheses. • What are the "omic" sciences ?. • From proteins to proteomics, metabolomics to the metabolites and lipids to Lipidomics. • Relationship of proteomics and other post-genomic science. technological platforms: • Preparation of the sample (plants, bacteria, animals): protein extract preparation for DIGE analysis, general guidelines for sample preparation for LC-MS analysis. • protein expression analysis, metabolites and lipids by electrophoretic techniques or Applications of 'SDS-PAGE and dell'IEF proteomics, metabolomics and lipidomics. or Differential Gel Electrophoresis (DIGE), 2D-DIGE saturation or capillary electrophoresis (CE), two-dimensional electrophoresis (2DE). or Applications of 2-DE in proteomics: limits and perspectives. or Blue Native electrophoresis on polyacrylamide gel (Blue Native PAGE). or Clear Native PAGE (CN-PAGE). Electrophoresis or 'gel free' or Electrophoresis on cellulose acetate • Methods for post analysis disclosure: reverse coloring, fluorescent dyes, Image Analysis for multiplexing technologies • Methods of detection of post-translational modifications in mass spectrometry: or glycosylation, phosphorylation, nitration, carbonylation • protein expression analysis using chromatographic techniques: or proteomic approach Bottom-up or proteomic approach Middle-down Approach or top-down proteomics or Multidimensional Separations • Mass Spectrometry: introduction and concepts of resolution and accuracy o The most used sources in the field of proteomics: MALDI, ESI o The most used sources in Metabolomics: EI o The most used sources in the field lipidomic: ESI, MALDI or analyzers: quadrupole, TOF and ion trap, Orbitrap, hybrid • The tandem mass spectrometry (MS \ MS) quantitative methods: liquid chromatography system (LC), reverse phase (RP) coupled to mass spectrometry analysis. or strategies relative quantification based on labeling with stable isotopes (iTRAQ, SILAC spike-in, Super-SILAC and SILAC organisms, ICAT, O18) or relative quantification strategies without marking ( "label-free" methods: intensity-based and spectral-counting, calculation of the PAI and dell'emPAI). or proteomics, metabolomics and targeted quantitative Lipidomics: SRM hyphenated techniques: GC-MS, CE-MS, LC-MS \ MS, MudPIT
Anna Maria Timperio
(reference books)
T. Alberio, M. Fasano, P. Roncada "PROTEOMICA" EdiSES; I. Lavagnini, F. Magno, R. Seraglia e P. Traldi "Quantitative Applications of Mass Spectrometry (English Edition)" WILEY.
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