18O-based protein quantitation: influence of separation on quantitation outcome in analysis of Staphylococcus a. enterotoxins

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HAVLIŠ Jan ŽUPKOVÁ Marcela NOVÁKOVÁ Kateřina ZDRÁHAL Zbyněk

Rok publikování 2010
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Popis Food contamination by enterotoxins from Staphylococcus aureus is of intense interest from side of food microbiologists and biochemists. Our work is focused on enterotoxin H (SEH). One of the main aims of our work is to reliably determine the content of SEH in whole secretome, on a background of all other staphylococcal enterotoxins that are partially sequence homologous. Mass spectrometry driven protein quantitation seems to be the most selective way. Prior to the quantitation, proteins must be separated. As the proteins are most conveniently quantified via their tryptic peptides, the separation should ensure availability of a quantifiable tryptic peptide. Here we present a study on influence of different separation ways, namely polyacrylamide gel electrophoresis, reversed phase liquid chromatography and off-gel isoelectric focusation, on availability of quantifiable tryptic peptides of SEH for 18O-based protein quantitation routine.
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