Springer Protocols: Abstract: Characterization of O-Linked Saccharides on Glycoproteins

Characterization of O-Linked Saccharides on Glycoproteins

By: Roger K. Bretthauer²

Abstract

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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.

Affiliation(s): (2) Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN

Book Title: Pichia Protocols

Series: Methods in Molecular Biology | Volume: 389 | Pub. Date: Aug-08-2007 | Page Range: 107-118 | DOI: 10.1007/978-1-59745-456-8_8

Subject: Biotechnology

Key Words: O-Mannosylation - β-elimination - reductive labeling - 2-aminobenzamide - oligosaccharide - mannosidase - high-pressure liquid chromatography (HPLC) - size-exclusion chromatography

References

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