LAUSR

Protein aggregates are a potential risk factor for immunogenicity. The measurement, characterization, and control of protein aggregates in drug products are indispensable for the development of biopharmaceuticals, including therapeutic monoclonal antibodies (mAbs). In this study, Fcγ receptor (FcγR)-expressing reporter cell lines were used to analyze the FcγR-activation properties of mAb aggregates. Comparison of aggregates of mAbs harboring different IgG subclasses revealed that the FcγR-activation profiles of the mAb aggregates were dependent on IgG subclass. In addition, aggregates of Fc-engineered mAb with enhanced FcγR-activation properties exhibited stronger activation of FcγRs than was observed in the wild type aggregates, whereas aggregates of Fc-engineered mAb with decreased FcγR-activation properties showed reduced activation. These results suggest that FcγR activation by mAb aggregates depends greatly on the Fc functions of the native (non-aggregated) mAbs. We also showed that aggregates of mAbs smaller than 1 μm in size have the potential to directly activate FcγRs. Unintended immune cell activation can be induced due to FcγR activation by mAb aggregates and such FcγR activation may contribute to immunogenicity, and therefore analysis of the FcγR-activation properties of mAb aggregates using FcγR-expressing reporter cell lines is a promising approach for the characterization of mAb aggregates.