Figure 6.

Ipilimumab (a) and mAb146 (b) bound to wide type human CTLA-4 and glycosylation sites mutated (N78Q and N110Q) CTLA-4. The N-glycosylation sites N78 and N110 on CTLA-4 monomer with His tag were removed by single mutation of N78Q, N110Q. These CTLA-4 variants and WT CTLA-4 were incubated with 1 μg/ml of antibodies pre-coated on a 96-well plate. The bound CTLA4 variants were detected by HRP-labeled secondary antibody. The absorbance at 450 nM was measured using a microplate spectrophotometer.