Immobilised enzyme reactors for post-production glycan modification of purified glycoproteins.
- 1Institute for Biomedicine and Glycomics, Griffith University, Gold Coast, QLD, Australia.
- 2ARC Centre of Excellence in Synthetic Biology, Macquarie University, Sydney, NSW, 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.
Related
- A numerically verified analytical framework for immobilised-enzyme packed-bed reactor design in the kinetically controlled regime applied to dihydroxyacetone production
- An efficient system for production of highly purified inhibins using mammalian cell expression and tandem affinity purification
- Advancement of mannosylerythritol lipids production: enhancement and tailor-made production through genetic modification of producing yeasts
- Differential Behavior of Uncleaved gp160 Human Immunodeficiency Virus Type 1 (HIV-1) Envelope (Env) Glycoproteins
- Palmitoylation of glycoproteins under low temperature enhances SVCV replication
BibTeX
@article{debono2026immobilised,
title = {{Immobilised enzyme reactors for post-production glycan modification of purified glycoproteins.}},
author = {Nicholas J DeBono and Joel Cain and Chi-Hung Lin and Nicolle H Packer and Edward S X Moh},
journal = {Frontiers in molecular biosciences},
year = {2026},
month = oct,
doi = {10.3389/fmolb.2026.1932584},
url = {https://europepmc.org/article/MED/42920519},
note = {CC BY},
}
RIS
TY - JOUR
TI - Immobilised enzyme reactors for post-production glycan modification of purified glycoproteins.
AU - Nicholas J DeBono
AU - Joel Cain
AU - Chi-Hung Lin
AU - Nicolle H Packer
AU - Edward S X Moh
PY - 2026
DA - 2026/10/09
JO - Frontiers in molecular biosciences
DO - 10.3389/fmolb.2026.1932584
UR - https://europepmc.org/article/MED/42920519
AB - 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.
SN - 2296-889X
ER -