Figure 5.

IgG-binding assay with CD16-transfected cells after blockade of glycosylation with tunicamycin. Cells transfected with WT-CD16B (a) and CD16-N163Q (b) were incubated for 30 hr with or without tunicamycin (0·5 μg/ml) and used for an IgG-binding assay afterwards. The cells were incubated with the indicated concentrations of human monomeric IgG (μg/ml) and subsequently with a FITC-labelled monoclonal antibody against human IgG. To take into account slightly different expressions of CD16, the MFI measured for each cell line and concentration of IgG (MFI X) was divided by the MFI of the reference antibody CLB-Gran1 on the respective cell line. All assays were performed in triplicate.