Site-specific glycosylation of an aglycosylated human IgG1-Fc antibody protein generates neoglycocoprotein with enhanced function

GM Watt, John Lund, Jacqueline Levens, K Kolli, Royston Jefferis, G-J Boons

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

A range of well-defined IgG glycoforms was prepared by employing a combination of synthetic carbohydrate chemistry and genetic engineering. The key aspect of this methodology is the coupling of thioaldoses with cysteine-containing proteins to give disulfide-linked neoglycoproteins. This technology was applied to the synthesis of a series of synthetic N-glycan thioaldoses which were coupled to an aglycosylated IgG1-Fc fragment, engineered to have Cys-297 in place of glycan-linked Asn (Deltah-Fc N297C). Analysis of the resulting Fc neoglycoproteins by mass spectrometry and trypsin digestion showed that the saccharides were site-selectively incorporated at Cys-297 to full occupancy without affecting other Fc protein disulfides. The neoglycoproteins were tested for their ability to interact with human FcgammaRI by inhibiting superoxide production by gamma-interferon-stimulated U937 cells. The neoglycoproteins displayed enhanced superoxide inhibition relative to aglycosylated Deltah-Fc N297C, where increased glycan size correlated positively with increased inhibition.
Original languageEnglish
Pages (from-to)807-814
Number of pages8
JournalChemistry & Biology
Volume10
Issue number9
DOIs
Publication statusPublished - 1 Sep 2003

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