The TSKgel FcR-IIIA columns from Tosoh Bioscience separate therapeutic antibodies based on the affinity of their Fc region for a particular Fc receptor (FcγIIIa). Responsible for the separation is a recombinant version of the FcγIIIa receptor, which is coupled to the stationary phase of the HPLC columns. FcγIIIa receptor affinity of a monoclonal antibody (mAb) is directly related to glycosylation and antibody-dependent cellular cytotoxicity (ADCC) activity. Thus, the columns rapidly link FcγIIIa receptor affinity with mAb function and structure. Additionally, they allow for characterizing different FcγIIIa receptor affinity fractions with orthogonal methods to investigate structural causes and functional effects of FcγIIIa receptor affinity.
Current chromatographic analyses of therapeutic antibodies focus on detecting structural differences related to functional and safety impairments. In contrary, FcR-IIIA affinity chromatography separates according to a characteristic directly linked to antibody function: affinity to the FcγIIIa receptor. The separation results correlate with glycosylation patterns and antibody-dependent cellular cytotoxicity (ADCC) activity, suggesting that the novel and unique methodology can complement and partially replace current analytical approaches.