Evaluation of mAb ADCC Activity through Fc Receptor Affinity Chromatography

Antibody-dependent cell-mediated cytotoxicity (ADCC) is a mechanism of action of anti-tumor antibodies in which the Fcy receptor (FcR) plays a key role. Hence, affinity chromatography using recombinant FcR can deliver valuable information about ADCC activity. Data on the analysis of therapeutic mAbs with the new TSKgel FcR-IIIA-NPR HPLC column are provided.

The N-glycans of antibody Fc domains are known to play an important role in Fc-mediated effector functions. Accordingly, separation patterns of therapeutic antibodies on TSKgel FcR-IIIA-NPR can be correlated to mAb glycoforms. Early eluting peaks (low affinity to Fc receptor) represent mAb glycoforms with low ADCC activity while late eluting peaks represent glycoforms with high ADCC activity.

Chromatographic conditions:

Column: TSKgel FcR-IIIA-NPR (5 µm 4.6 x 75 mm); Mobile Phase:  A: 50 mM Citrate, pH 6.5; B: 50mM Citrate, pH 4.5; Flow rate: 1 mL/min; Detection: UV@280 nm; Sample: Rituximab

Read the full application note here.

FcR-IIIA affinity chromatography separates mAb glycoforms according to their ADCC activity

Newsletters

Receive the latest pathologist news, personalities, education, and career development – weekly to your inbox.

I have read and understand the Privacy Notice*

Subscribe