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Improved Mass Spectrometric Proteomic Profiling of the Secretome of Rat Vascular Endothelial Cells

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journal contribution
posted on 06.10.2006, 00:00 by M. C. Pellitteri-Hahn, M. C. Warren, D. N. Didier, E. L. Winkler, S. P. Mirza, A. S. Greene, M. Olivier
Serum albumin contamination of cells cultured in vitro significantly impedes the mass spectrometric analysis of proteins secreted by the cells. Here we report a novel washing and culturing technique for rat vascular endothelial cells that considerably reduces the concentration of the commonly used additive for cell culture, bovine serum albumin (BSA), in the secretome of these cells. Cells are rinsed stringently and cultured for 24 h in serum-free media without appreciably impeding cell growth or viability. The percentage of BSA scans identified by tandem mass spectrometry (LC−MS/MS) in stringently rinsed cells (average 13.2%) was significantly lower than either the moderately rinsed or no rinse cell treatments (average 35.2% and 45.2% respectively). Furthermore, the stringent wash treatment allowed the confident identification of a larger portion of the secretome of rat endothelial cells by LC−MS/MS. Keywords: secretome • serum removal • liquid chromatography mass spectrometry • rat endothelial cell

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