Prof. Andras Guttman


The Analytical Challenges of Glycomics



Comprehensive analysis of protein glycosylation represents one of the most challenging
bioanalytical tasks as glycans are very complex groups of molecules. The lack of chromophore /
fluorophore moieties and, in many instances, easily ionizable groups usually require derivatization
of carbohydrates before their analysis by high performance bioanalytical techniques, such as liquid
chromatography, electrophoresis and mass spectrometry. Full structural elucidation of glycans also
utilizes consecutive enzymatic digestion by and array of exoglycosidases, followed by LC or
capillary electrophoresis separation of the digests. Hyphenation with offline weak anion exchange
chromatography (WAX) fractionation is an additional powerful tool, especially to decipher highly
sialylated complex sugars. This presentation will cover the state of the art of liquid phase
separation methods for structural eluc idation of protein glycosylation mostly focusing on
capillary electrophoresis (CZE and CGE) but also including the main approaches of liquid
chromatography and microcfluidics methods, as well as their combination with mass spectrometry.
Particular attention will be paid to the analysis of recombinant glycoprotein therapeutics (both
innovative and biosimilars), a recently emerging class of very successful new generation drugs with
the main goal to demonstrate structural and functional equivalence of the products. Comprehensive
characterization of their glycosylation is crucial in the biotechnology and biopharmaceutical
industry, especially in clone selection, process development and lot release.