Analysis of glycoproteins and the oligosaccharides thereof by high‐performance capillary electrophoresis—significance in regulatory studies on biopharmaceutical products

Abstract

This review describes the recent development in the analysis of glycoproteins using capillary electrophoresis with various separation techniques, and focuses especially on the analysis of recombinant glycoprotein pharmaceuticals. We include the analysis of glycoprotein multiforms (ie glycoform) as well as glycan analysis. The relationship between glycoprotein multiforms and oligosaccharide distributions in a glycoprotein sample is also discussed. Further, recent development in capillary electrophoresis-mass spectrometry is described.

Acute Phase Proteins: Structure and Function Relationship

“…The changes comprise alterations in branching pattern as revealed by reactivity with concanavalin A and the fucose-binding lectin (Elliott et al, 1997). Thus analysis of sialo-and asialo-oligosaccharides of AGP as well as its glycoforms is important for understanding the biological roles of AGP (Kakehi et al, 2002;Sei et al, 2002). For example, the asialylated carbohydrate-deficient variant of AGP appears mainly in sera of patients after acute inflammation, infection, burns or other severe tissue damage (Fournier et al, 2000).…”

Chromatographic and electrophoretic characterization of protein variants

“…One major application is the characterization of glycoprotein variants [137,138]. A set of different microscale techniques of CE exists for analyzing recombinant glycoproteins.…”

Electrophoretic analysis of di- and oligosaccharides derived from glycosaminoglycans on microchip format

“…In the carbohydrate analyses, Tris-borate buffer often shows better separation efficiency than Tris-acetate buffer probably due to competitive complex formation between carbohydrates/Tris and borate ions. Low-operation voltage in Tris-borate buffer is also preferable for electrophoresis [8].…”

“…Capillary electrophoresis has revealed the ability in separation of carbohydrates, although most carbohydrates must be converted to ion forms prior to the analysis. Furthermore, carbohydrates should be labeled with chromophores or fluorophores for sensitive detection [8].…”