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Application of affinity chromatography and affinity-mass spectrometry in protein identification

Autoři: Svobodová Eliška | Jankovičová Barbora | Bílková Zuzana | Przybylski M.

Rok: 2007

Druh publikace: ostatní - přednáška nebo poster

Strana od-do: nestránkováno

Tituly:

Jazyk	Název	Abstrakt	Klíčová slova
cze	Aplikace afinitní chromatografie a afinitní hmotnostní spektrometrie v identifikaci proteinů.	Aplikace afinitní chromatografie a afinitní hmotnostní spektrometrie v identifikaci proteinů.	ESI MS, MALDI MS, proteolytická excision, peptidová extraction
eng	Application of affinity chromatography and affinity-mass spectrometry in protein identification	Affinity chromatography is a powerful technique for protein or peptide separation based on the specific interaction between immobilized ligand and target protein [1]. The interactions that occur in this method are the same that take place in many biological systems, i. e. the binding of an antibody with an antigen or of an enzyme with a substrate. The nature of these interactions makes affinity chromatography a high selective separation and purification method and makes it possible to isolate only a certain analyte or group of analytes from complex samples. Usually proteome analysis consists of protein isolation by two-dimensional electrophoresis and identification of a set of proteolytic fragments by mass spectrometry. Affinity chromatography is used within these procedures for simplification of protein or peptide mixtures complexity. Peptide mapping is a convenient and widely used method for the identification and characterization of proteins. This technique consists of site-specific enzymatic digestion of the protein followed by separation using affinity chromatography and detection of the produced peptides using ESI and MALDI mass spectrometry [2]. Proteolytic peptide excision and peptide extraction following affinity-binding to an immobilised antibody are important techniques of protein identification (affinity-mass spectrometry [3]) used recently for identification of diagnostic markers. Affinity mass spectrometry of specific proteins as well as of protein complexes followed by enzymatic digestion serves as an effective tool for identification of proteins as will be shown by recent examples of carbohydrate-binding proteins. [1] AZARKAN M., HUET J., BAEYENS-VOLANT D., LOOZE Y., VANDENBUSSCHE G.: Affinity chromatography: A useful tool in proteomics studies. J. Chromatogr. B (2006), doi:10.1016/j.jchromb.2006.10.056. [2] LEE W.-CH., LEE K. H.: Application of affinity chromatography in proteomics. Anal. Biochem., 324 (2004): 1 - 10. [3] MACHT M., MARQUARDT A.	ESI MS; MALDI MS; proteolytic excision; peptide extraction

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