Figure 7
From: Multi-level glyco-engineering techniques to generate IgG with defined Fc-glycans
Increasing sialylation.
(a) The sialylation level of IgG1 N297 after co-transfection of IgG heavy and light chain vectors with 1% B4GALT1 and addition of 5 mM D-galactose to increase galactosylation, in combination with increasing amount of co-transfected ST6GALT. N = 2 (b) Effect of ST6GALT co-transfection on other derived glycosylation traits. N = 2 (c) In vitro sialylation by ST6GALT of highly galactosylated and sialylated IgG1. Before treatment N = 3, after treatment N = 7 (d) NanoLC-ESI-MS spectrum of IgG produced with the identified optimal co-transfection additions of B4GALT1 (1%) and ST6GALT (2,5%) with the addition 1 mM D-galactose, and (e) spectrum of in vitro sialylated IgG produced in (d). The data represents means and SD, **** denotes a statistical significance of p ≤ 0.0001 tested by one-way ANOVA using Dunnett’s multiple comparisons test (a,b) or unpaired t-test (c) comparing untreated cells or IgG respectively with treated. The vertical dotted lines in (a,b) represent the designated optimal concentrations of ST6GAL.
