Gel-free analysis of the human brain proteome: Application of liquid chromatography and mass spectrometry on biopsy and autopsy samples
Dumont, Debora × Noben, Jean-Paul Verhaert, Peter Stinissen, Piet Robben, Johan #
Wiley-v c h verlag gmbh
Proteomics vol:6 issue:18 pages:4967-4977
This paper reports on the findings of the Biomedical Research Institute, as one of the participants in the pilot study of the HUPO Brain Proteome Project. A biopsy and autopsy study sample derived from human brain was distributed among the participants for proteomic analysis. In our laboratory, attention was focused on protein identification using the bottom-up shotgun approach. Protein extracts derived from both samples were trypsinized and analyzed separately by 2-D LC and MS. In a complementary approach, the tryptic digests were analyzed directly by LC-ESI-MS/MS and gas-phase fractionation in the mass spectrometer. Taken together, both proteomic approaches in combination with a stringent evaluation process, resulted in the confident identification of 209 proteins in the human brain samples under investigation.