Sialylated carbohydrate chains of recombinant human glycoproteins expressed in Chinese hamster ovary cells contain traces of N‐glycolylneuraminic acid

[摘要]

HPLC analysis of sialic acid released from recombinant variants of human tissue plasminogen activator, human chimeric plasminogen activator, human erythropoietin, and human follitropin, expressed in Chinese hamster ovary cells, demonstrates for each glycoprotein the presence of N-acetylneuraminic and N-glycolylneuraminic acid in a ratio of 97:3. Structural analysis by 500 MHz¹H-NMR spectroscopy, of the enzymatically released N-linked carbohydrate chains of chimeric plasminogen activator and of erythropoietin, showed that α2-3 linked N-glycolylneuraminic acid can occur in different N-acetyllactosamine type antennary structures.

[学科分类] 生物化学/生物物理

[关键词] Recombinant human glycoprotein;Chinese hamster ovary cell;N-Glycolylneuraminic acid;Neu5Ac;N-acetylneuraminic acid;Neu5Gc;N-glycolylneuraminic acid;Fuc;fucose;CHO;Chinese hamster ovary;tPA;tissue plasminogen activator;k2tuPA;recombinant chimeric plasminogen activator;EPO;erythropoietin;FSH;follicle stimulating hormone;follitropin;hCG;human chorionic gonadotropin;PNGase-F;peptide-N 4-(N-acetyl-β-glucosaminyl)asparagine amidase F;DMB;1;2-diamino-4;5-methylene-dioxybenzene