• YAKHAK HOEJI
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• v.53 no.1
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• pp.34-40
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• 2009

The glycosylation of therapeutic glycoproteins can affect their efficacy, stability, solubility, and half-life. Analyzing the composition of monosaccharides, such as that of neutral and amino sugars, is the first step for elucidating the structure of glycan attached to glycoproteins. In the present study, neutral and amino sugars of lectin obtained from Maackia fauriei 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. The hydrosylate of lectin was prepared by treatment with trifluoroacetic acid, which resulted in the lectin mainly possessing the N-glycan consisting of 98.1 pmol Fuc, 342.1 pmol GlcN, 51.9 pmol Gal, 678.9 pmol Man, and 330.7 pmol Xyl. The present results demonstrate that ELLA and HPAEC-PAD are very effective methods for rapidly estimating the types and relative amounts of monosaccharides in intact glycoproteins.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.248.2-248.2
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• 2003

A lectin has been purified from the bark of the legume Maackia fauriei. This lectin, MFA, was found to agglutinate human ABO erythrocytes at a titer of 256. The results from electrophoretic analyses, gel-filtration chromatography, and enzyme linked lectinsorbent assay indicate that MFA is an acidic glycoprotein, and exists as a tetramer of 30 kDa subunits that are linked by noncovalent bonds. The activity of MFA is critically dependent upon CaC1$_2$. MFA demonstrated high homogeneity with the lectins from M. amurensis, which is the only legume source of lectins that bind to sialoglycoconjugate, in its N-terminal amino acid sequence and amino acid composition, (omitted)