TITLE:
Validation of an Easy Acetonitrile Fractionation for the Simplification of Protein Samples Prior to Proteomics Analyses
AUTHORS:
Yannis Karamanos, Barbara Deracinois, Gwënaël Pottiez, Sophie Duban-Deweer
KEYWORDS:
Proteomics, Acetonitrile, Fractionation
JOURNAL NAME:
Open Access Library Journal,
Vol.2 No.10,
October
14,
2015
ABSTRACT: In most proteomics analyses and in particular for the “off-gel”
approaches, based essentially on chromatography, the complexity of the proteome
should be reduced; otherwise identifications can be hindered, especially if the
mass spectrometry analysis is not conducted using state-of-the-art
instrumentation. Even if the method used is a bottom-up proteomics, it appears
mandatory to pre-fractionate the proteins in order to reduce the complexity. We
report here the development and validation of a pre-fractionation based on the
differential solubilisation of proteins using increasing concentrations of
acetonitrile (ACN). This “ACN fractionation” was applied to the study of the
Triton X-100 soluble sub-proteome of brain capillary endothelial cells (BCEC)
with re-induced blood-brain barrier (BBB) functions.