Quantitative assessment of mAb Fc glycosylation of CQA importance by capillary electrophoresis

Abstract

The attached carbohydrates at the highly conserved asparagine-linked glycosylation site in the C_H 2 domain of the fragment crystallizable (Fc) region of monoclonal antibody therapeutics can play an essential role in their mechanism of action, including ADCC, CDC, anti-inflammatory functions, and serum half-life. Thus, this particular glycosylation represents one of the important critical quality attributes (CQA) of therapeutic monoclonal antibodies, which should be closely monitored and controlled during all stages of biopharmaceutical manufacturing. To study Fc glycosylation related quantitative critical quality attributes, the N-glycan pool of adalimumab (Humira^® ) was spiked with increasing amounts of mannose-5 oligosaccharide, a glycan with high CQA importance. The method enabled precise quantitative CQA assessment with high detection sensitivity.

Keywords: Biotherapeutics; Capillary electrophoresis; Critical quality attributes; Glycosylation; Quantification.