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Application of Mass Spectrometry in Proteomics
Authors:Ida Chiara Guerrera  Oliver Kleiner
Institution:(1) Department of Medicine, Centre for Molecular Medicine, University College London, 5 University Street, WC1E 6JJ London, UK;(2) The Rayne Institute, University College London, 5 University Street, WC1E 6JJ London, UK
Abstract:Mass spectrometry has arguably become the core technology in proteomics. The application of mass spectrometry based techniques for the qualitative and quantitative analysis of global proteome samples derived from complex mixtures has had a big impact in the understanding of cellular function. Here, we give a brief introduction to principles of mass spectrometry and instrumentation currently used in proteomics experiments. In addition, recent developments in the application of mass spectrometry in proteomics are summarised. Strategies allowing high-throughput identification of proteins from highly complex mixtures include accurate mass measurement of peptides derived from total proteome digests and multidimensional peptide separations coupled with mass spectrometry. Mass spectrometric analysis of intact proteins permits the characterisation of protein isoforms. Recent developments in stable isotope labelling techniques and chemical tagging allow the mass spectrometry based differential display and quantitation of proteins, and newly established affinity procedures enable the targeted characterisation of post-translationally modified proteins. Finally, advances in mass spectrometric imaging allow the gathering of specific information on the local molecular composition, relative abundance and spatial distribution of peptides and proteins in thin tissue sections.
Keywords:Mass spectrometry  proteomics  top-down and bottom-up approaches  relative and absolute protein quantitation
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