Capillary electrophoresis of glycans: Validation of antibody pharmaceuticals based on glycan profiles

Abstract

The N-glycosylation on therapeutic monoclonal antibodies (mAbs) plays important biological roles such as antibody-dependent cellular cytotoxicity (ADCC) or circulatory half-time in vivo. Although the attached N-glycans are mostly core α1-6-fucosylated biantennary glycans, minor nonhuman N-glycans having N-glycolylneuraminic acid (NeuGc) and Galα1-3Gal epitopes (αGal) have been reported to be present in therapeutic mAbs. Determination of nonhuman N-glycans is important in the evaluation of the mAb characteristics, because the presence of nonhuman N-glycans may affect the safety of mAb products leading to immunogenicity or adverse incident. In this chapter, we present the method for validation of mAbs based on glycan profiles using capillary electrophoresis (CE) with laser-induced fluorescence detection (LIF). The presented method using CE-LIF allows the routine testing to ensure safety and efficacy of the mAb products.