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Mass spectrometry-based proteomics – evolving technology in the field of medicine

The emerging need for systems-based approach introduced application of proteomic technologies to the field of medicine. MS-based proteomics is nowadays employed for large-scale protein identification in vitro, in vivo or in clinical research to determine pathways, subcellular protein distributions, cellular functions, protein interaction maps, posttranslational modifications (e.g. glycosylation, phosphorylation), and diagnostic/prognostic biomarkers. It is also applied to quantitative protein analyses. Proteins can be identified according to bottom up gel- or liquid chromatography (LC)-based proteomic protocols. For that purposes two major MS platforms based on the ionization type are employed: matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI). Generally, MALDI is more related to gel-based, while ESI is applied in LC-based proteomics. In gel-based proteomic approach, after protein extraction from the biological samples, complex protein mixtures are resolved by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Deregulated protein spots are excised, digested in gel, and resulting peptides are analyzed by MS. Tandem mass spectrometry coupled to liquid chromatography (LC-MS/MS) allows protein identification in a complex mixtures after protein digestion in solution without need for previous protein purification (approach known as "shotgun" proteomics). Digested proteins are separated by LC and analyzed by MS in order to produce mass and tandem mass spectra. Obtained data are used for bioinformatics analyses, such as database search for protein identities, gene ontology analysis, and interactome. Systems-based approach, technical advances in LC-based identification and quantification, especially in targeted analyses, and sophisticated bioinformatic algorithms made mass spectrometry-based proteomics a powerful tool in diagnostics and biological research.

mass spectrometry; proteomics; 2-D gel; LC-MS

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