Differential influence on antibody dependent cellular phagocytosis by different glycoforms on therapeutic Monoclonal...

Therapeutic monoclonal antibodies (mAbs), particularly of the IgG1 subclass, are capable of effector function activities that may be important for their mechanism of action. One such effector function activity is Antibody Dependent Cellular Phagocytosis (ADCP), which has been shown to be mediated primarily through the activating FcγR, FcγRIIa, on macrophages and neutrophils. The critical quality attributes that are the most impactful and predictive of ADCP activity, and therefore most suitable to monitor during IgG1 antibody manufacturing, are not well established. Primary cell assays for ADCP are often laborious and subject to donor to donor variability, making such assays less desirable for product characterization. By developing and employing an ADCP reporter gene assay, we have been able to determine with high sensitivity the glycan structures that can impact FcγRIIa mediated ADCP across multiple different IgG1 antibodies. Interestingly we observed that some IgG1 antibodies are very potent mediators of ADCP while others do not mediate ADCP even though they possess other effector function activities (ADCC and CDC). Additionally, we find that ADCP by different IgG1 antibodies has markedly different sensitivity to glycan species, with one antibody demonstrating a surprisingly strong influence of β-galactosylation and high mannose levels.