{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T04:50:29Z","timestamp":1791521429981,"version":"4.3.4"},"reference-count":60,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T00:00:00Z","timestamp":1791504000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Mol. Biosci."],"abstract":"<jats:p>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 \u03b2-1,4-galactosyltransferase (B4GalT1-IMER) and \u03b1-2,6-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.<\/jats:p>","DOI":"10.3389\/fmolb.2026.1932584","type":"journal-article","created":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T04:27:05Z","timestamp":1791520025000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Immobilised enzyme reactors for post-production glycan modification of purified glycoproteins"],"prefix":"10.3389","volume":"13","author":[{"given":"Nicholas J.","family":"DeBono","sequence":"first","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University","place":["Gold Coast, QLD, Australia"]},{"name":"ARC Centre of Excellence in Synthetic Biology, Macquarie University","place":["Sydney, NSW, Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joel","family":"Cain","sequence":"additional","affiliation":[{"name":"ARC Centre of Excellence in Synthetic Biology, Macquarie University","place":["Sydney, NSW, Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chi-Hung","family":"Lin","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University","place":["Gold Coast, QLD, Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolle H.","family":"Packer","sequence":"additional","affiliation":[{"name":"Institute for Biomedicine and Glycomics, Griffith University","place":["Gold Coast, QLD, Australia"]},{"name":"ARC Centre of Excellence in Synthetic Biology, Macquarie University","place":["Sydney, NSW, Australia"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Edward S. 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