Proteomics enabled Vaccinology. Probing Antigen and Epitope Repertoires

摘要: The immune system is a complex of molecules, cells, tissues and organs that protects higher organisms from pathogens or cancerous cells. A proper understanding the vital for development new vaccines against infectious diseases immunotherapies cancer. Mass Spectrometry-based proteomics an important analytical strategy in vaccinology enables identification quantification proteins cells tissue, under normal treated (diseased) conditions. This thesis covers major methodological aspects workflow, sample enrichment to peptide identification. Applied different research questions these methods address challenges complexity sensitivity, as difficulties sequencing. positional was introduced selective isolation N-terminal peptides. method effectively reduces proteomes global characterization post translation modifications. applied quantitatively profile protein content Neisseria Meningitidisouter membrane vesicles (OMV)-based vaccines. Differences were found between OMV produced purification quantitative data sustained by serum blot proteomics, confirming differential pattern antigenic content. furthermore describes two dimensional (2D) chromatographic separation technique highly sensitive analysis mixtures. employs ion exchange chromatography reversed phase online configuration. introduction novel buffer omitted use undesirable salts, resulting excellent compatibility 2D liquid mass spectrometry. Another part utility recently fragmentation Human Leukocyte Antigens (HLA) class I-associated dual generate both fragment ions induced Electron-Transfer Dissociation (ETD) Higher-energy collision (HCD) single spectrum. endogenous peptides considerably benefits complementary informative-rich spectra provided this approach. Novel features antigen processing machinery are reported, including variety translational modifications which evidence accumulating they play roles human diseases.