Fig. 3.

IgG glycan hydrolysis abolishes ADCC. (A) Illustration of the iLite® ADCC assay (Svar Life Science). Target cells expressing the antigen are combined with the mAb of interest and reporter cells expressing FcγRIII (CD16). The reporter cells further express a luciferase induced by CD16 signaling, leading to an easy read-out for activation of ADCC. (B) ADCC signal of the monoclonal antibodies trastuzumab (left) and rituximab (right) as determined by the iLite® ADCC assay. The signals elicited by the untreated, fully glycosylated mAbs is shown in blue, the signal from the endoglycosidase-treated mAbs in red. Note the almost complete abolishment of CD16 signaling upon IgG glycan hydrolysis.