Control of IgG/Fc glycosylation : a comparison of oligosaccharides from chimeric human/mouse and mouse subclass immunoglobulin Gs

摘要: Abstract Oligosaccharide profiles were obtained for chimeric mouse-human antibodies corresponding to each of the human IgG subclasses 1–4, and mouse IgG2b expressed in J558L cell line. These have specificity NIP hapten form a matched set IgGs. An IgG4 antibody (B72.3) produced Chinese hamster ovary (CHO-K1) line was also analysed carbohydrate. Additionally aglycosylated mutants this anti-NIP analysed. The total lack carbohydrate found site-directed B72.3 (Asn 297→ Gln) 297 → Ala) demonstrates that there are no N-glycosylation sites other than Asn 297. Therefore glycosylation all IgGs reflect attached site. Factors such as type (A), template direction by heavy chains (B) culture conditions (C) shown influence profiles. (A) may one or two Gal (alpha 1→ 3) residues per oligosaccharide unit, indicative presence galactosyl transferase galactosylation chains, particular preference Man 6) arm rather arm, contrary beta-galactosyltransferase specificity, suggest polypeptide chain act extent hence proportions incorporated. IgG2 does not display preference. appears be influenced growth conditions, with highest levels being cells grown still cultures, ascites hollow fibre bioreactors. It is concluded though profile controlled predominantly activity which expressed, differences between templates various can glycosylation.