Exploring the proteome by CE-ESI-MS

Abstract

Recent developments in the coupling of liquid phase separations and mass spectrometry have yielded some new source designs for electrospray ionization. The research in this thesis covers the application of a newly developed electrospray ionization source design and it's application for the coupling of capillary electrophoresis to mass spectrometry. The initial chapters cover improvements in sensitivity and separation power in capillary electrophoresis mass spectrometry (CE-MS) by allowing electrospray at ultra low flow rates and therefore optimal ionization and separation conditions. We showed strong complementarity of this CE-MS system with reversed phase liquid chromatography in the identification of peptides and proteins from e.coli digests. Finally a sample preparation method for peptide and protein identification from laser micro-dissected tissue samples from human kidneys was developed. It was shown that a few hundred proteins could be identified from amounts of material representative of one full human glomerulus. On the whole great strides were made in development of strategies employing CE-MS for proteomics analysis.