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Updated: Oct 25, 2026

Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases
Published on: December 26, 2011
Immobilised enzyme reactors for post-production glycan modification of purified glycoproteins
Nicholas J DeBono1,2, Joel Cain2, Chi-Hung Lin1
1Institute for Biomedicine and Glycomics, Griffith University, Gold Coast, QLD, Australia.
Abstract:
Controlling protein glycosylation as a critical quality attribute of biopharmaceuticals remains challenging when glycosylation is coupled to cellular production systems. Here, we present a proof-of-concept glycosyltransferase immobilised enzyme reactor (IMER) housed within a 3D-printed column that enables directed post-production glycan modification of purified glycoproteins. Using β-1,4-galactosyltransferase (B4GalT1-IMER) and α-2,6-sialyltransferase (ST6Gal1-IMER) immobilised on Ni-NTA resin, the IMER achieved near-complete galactosylation and substantial sialylation of partially de-glycosylated bovine fetuin N-glycans with their respective substrates with a maximum substrate-enzyme contact time of 4 minutes. Isomeric-level analysis revealed arm-specific addition preferences for both enzymes, consistent with known specificities. The modular IMER design permits sequential connection of individual enzyme chambers, potentially offering a scalable, plug-and-play platform for constructing defined glycan structures on recombinant glycoprotein therapeutics.
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