Enrichment and characterization of glycopeptide epitopes from complex mixtures

Antigen posttranslational modifications, including glycosylation, are recognized by the innate and adaptive arms of the immune system. Analytical approaches, including mass spectrometry and allied techniques, have allowed advances in the enrichment and identification of glyco-antigens, particularly T-cell epitopes. Similarly, major advances have been made in the identification, isolation, and detailed characterization of prokaryotic and eukaryotic glycoproteins and glycopeptides. In particular, peptide centric approaches are now capable of enriching low level glycopeptides from highly complex peptide mixtures. Similarly, advanced mass spectrometry methods allow identification of glycopeptides, characterization of glycans, and mapping of modification sites. Herein, we describe methods developed in our laboratory for the broad study of glycopeptides and illustrate how these approaches can be exploited to further our understanding of the identity and nature of glycopeptide epitopes in various diseases or auto immune disorders.