Work

Immobilised enzyme reactors for post-production glycan modification of purified glycoproteins

Year: 2026

Type: article

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.

Source: Frontiers in Molecular Biosciences

DOI: 10.3389/fmolb.2026.1932584

Authors: Nicholas J. DeBono, Joel Cain, Chi-Hung Lin, Nicolle H. Packer, Edward S. X. Moh

Institutions: Griffith University, ARC Centre of Excellence in Synthetic Biology, Macquarie University

Language: English

Topic: Glycosylation and Glycoproteins Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $3,801

Funders: Commonwealth Scientific and Industrial Research Organisation, Macquarie University, National Institute of General Medical Sciences