Capillary electrophoresis for the analysis of glycoprotein pharmaceuticals

Abstract

Carbohydrate chains in glycoprotein pharmaceuticals play important roles for the expression of their biological activities, but the structure and compositions of carbohydrate chains are dependent on the conditions for their production. Therefore, evaluation of the carbohydrate chains is quite important for productive process development, characterization of product for approval application, and routine quality control. The oligosaccharides themselves have complex structure including blanching and various glycosidic linkages, and oligosaccharides in one glycoprotein pharmaceutical generally have high heterogeneity, and characterization of oligosaccharide moiety in glycoprotein has been a challenging target. In these situations, CE has been realized as a powerful tool for oligosaccharide analysis due to its high resolution and automatic operating system. This review focuses on the application of CE to the glycoform analysis of glycoproteins and profiling of the N-linked glycans released from glycoprotein pharmaceuticals. Current applications for structure analysis using CE-MS(n) technique and glycan profiling method for therapeutic antibody are also described.

Capillary electrophoresis of proteins 2005–2007

“…CZE in drug analysis including proteins has been reviewed recently [147] as well as CZE of carbohydrate chains in glycoproteins [148]. CE-MS has been used to characterize and study therapeutic proteins, including profiling N-linked glycopeptide heterogeneity and determining O-linked site occupancy [149].…”

Evaporative fluorophore labeling of carbohydrates via reductive amination

“…Glycosylation is one of the major and most complicated forms of post-translational modifications that plays essential roles in the activity and effector function of monoclonal antibodies [3] demanding improved glycoanalytical methods. Frequently 3/13 used techniques for carbohydrate analysis are mass spectrometry (MS) [4], liquid chromatography (LC) [5,6], capillary electrophoresis (CE) [7][8][9], some combination of them [10][11][12] and if necessary, nuclear magnetic resonance spectroscopy (NMR) [13,14].…”

Evaluation of glycosylation for quality assurance of antibody pharmaceuticals by capillary electrophoresis

“…In addition, the oligosaccharides corresponding to G0-G2, which do not have a fucose residue at the reducing terminal, can be clearly observed at faster migrating times (Fig. 7) [48]. In this case, the oligosaccharides are labeled with an analogous compound to 2-AA, 3-AA, which shows similar characteristics on the analysis of oligosaccharides by CE.…”