The TSKgel^® FcR-IIIA-5PW column is the second in the FcRγ affinity chromatography portfolio for antibody drug analysis. This column is a semi-preparative column based on a recombinant FcγRIIIA receptor ligand bonded to porous 10 µm polymethacrylate particles. The column can be loaded with up to 5 mg of mAb, allowing material collection in sufficient quantity for in-depth analysis of monoclonal antibody (mAb) glycoforms.

Like the TSKgel FcR-IIIA-NPR column, the TSKgel FcR-IIIA-5PW separates monoclonal antibodies into 3 fractions of affinity to the FcγRIIIA ligand: low, medium and high affinity. These fractions correlate with different mAb glycoforms and their antibody-dependent cellular toxicity (ADCC) activity. These columns represent a tool to rapidly link FcγIIIa receptor affinity with mAb function and structure. Characterizing each of the fractions with orthogonal methods allows for linking FcγIIIa receptor affinity to structural causes and functional effects. For this purpose, a workflow was established to determine the glycosylation pattern in different affinity fractions with HILIC-MS.