earticle

영어

The glycosylation of therapeutic glycoproteins can affect their efficacy, stability, solubility, and half-life. Especially, the analysis of mono- and oligosaccharide on the glycoproteins are an important part of glycoprotein characterization, because sialylation or fucosylation in oligosaccharides often causes dramatic changes in the function of glycoproteins. Analyzing the composition of monosaccharides, such as that of neutral, amino sugars and sialic acids, is the first step for elucidating the structure of glycan attached to glycoproteins. In the present study, monosaccharides obtained from glycorproteins were analyzed using an enzyme-linked lectinsorbent assay(ELLA) and high-performance anion exchange chromatography with pulsed amperometric detection(HPAEC-PAD). Peroxidase-labeled lectins such as concanavalin A, Ricinus communis agglutinin, and soybean agglutinin were used for ELLA, since they specifically bind to the monosaccharide residue most frequently encountered in a glycan. We also prepared pyridylamino-oligosaccharides from therapeutic glycoproteins, and their structural analysis was performed using 3D mapping method. The present results demonstrate that ELLA, HPAEC-PAD, and 3D mapping are very effective methods for rapidly estimating the types and relative amounts of monosaccharides and structural analysis of oligosaccharide in intact glycoproteins.