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Göteborgs universitets publikationer

O-glycan repertoires on a mucin-type reporter protein expressed in CHO cell pools transiently transfected with O-glycan core enzyme cDNAs.

Författare och institution:
Jining Liu (Institutionen för biomedicin, avdelningen för klinisk kemi och transfusionsmedicin); Chunsheng Jin (Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi); Reeja Maria Cherian (Institutionen för biomedicin, avdelningen för klinisk kemi och transfusionsmedicin); Niclas G. Karlsson (Institutionen för biomedicin, avdelningen för medicinsk kemi och cellbiologi); Jan Holgersson (Institutionen för biomedicin, avdelningen för klinisk kemi och transfusionsmedicin)
Publicerad i:
Journal of biotechnology, 199 s. 77-89
Artikel, refereegranskad vetenskaplig
Sammanfattning (abstract):
Glyco-engineering of host cells is used to increase efficacy, decrease immunogenicity and increase circulatory half-lives of protein biopharmaceuticals. The effect of transiently expressed O-glycan core chain glycosyltransferases on O-glycan biosynthesis pathways in CHO cells is reported. Liquid chromatography-mass spectrometry and Western blotting were used to map the O-glycome of a mucin-type fusion protein transiently co-transfected with β1,3-N-acetylglucosaminyltransferase 3 (extended C1 β3GnT3), core 2 β1,6-N-acetylglucosaminyltransferase I (C2 β3GnT1) or core 3 β1,3-N-acetylglucosaminyltransferase 6 (C3 β3GnT6) in CHO cells. Extended core 1 (GlcNAcβ1,3Galβ1,3GalNAc) and core 3 (GlcNAcβ1,3GalNAc), and increased expression of core 2 [Galβ1,3(GlcNAcβ1,6)GalNAc], O-glycans were generated on P-selectin glycoprotein ligand-1/mouse IgG2b (PSGL1/mIgG2b). Endogenous poly-N-acetyllactosamine (poly-LacNAc) synthase elongated extended core 1 and core 3 generating O-glycans with up to five LacNAc repeats. Low amounts of core 3 O-glycans appeared upon extended C1 β3GnT3 expression. The α2,6-sialylated type 2 chain was detected upon co-transfection with the β-galactoside α2,6-sialyltransferase I. N-acetylglucosamine-6-O-sulfotransferase 2 transferred sulfate to carbon 6 of GlcNAc in poly-LacNAc sequences. CHO cells with its known O-glycan repertoire can be used to express recombinant mucin-type proteins together with selected glycosyltransferases in order to recreate carbohydrate determinants on defined O-glycan chains. They will become important tools for assessing the core chain-dependent binding activity of carbohydrate-binding proteins.
Ämne (baseras på Högskoleverkets indelning av forskningsämnen):
Medicinsk bioteknologi
Postens nummer:
Posten skapad:
2015-04-08 17:28
Posten ändrad:
2016-08-22 13:19

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