Three-dimensional mapping of N-linked oligosaccharides using anion-exchange, hydrophobic and hydrophilic interaction modes of high-performance liquid chromatography |
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Authors: | Noriko Takahashi |
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Affiliation: | GlycoLab, Nakano Vinegar Co. Ltd., 2-6 Nakamura-cho, Handa-City 475, Japan |
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Abstract: | To determine the structure of N-linked oligosaccharides, a three-dimensional (3-D) sugar mapping technique for pyridylaminated neutral and sialyl oligosaccharides is proposed. The pyridylaminated oligosaccharide mixture is first separated by HPLC on a diethylaminoethyl anion-exchange column and the elution data are placed on the Z-axis. Neutral and mono-, di-, tri- and tetrasialyloligosaccharides are then individually separated on both a hydrophobic octadecylsilylsilica column and a hydrophilic amide-silica column under the same conditions as described previously for neutral oligosaccharides. The validity of the 3-D mapping technique was tested using sialyl pyridylaminated oligosaccharides from human serum glycoproteins. |
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