Characterization of O-Linked Saccharides on Glycoproteins

Recombinant and native proteins of Pichia pastoris can be O-mannosylated on serine and threonine residues, allowing further elongation reactions to generate short O-linked oligosaccharides of mannose. Methods for release from the protein with alkaline β-elimination with or without reduction of the released saccharides, and for subsequent chromatographic and enzymatic characterization of these saccharides are described.