Affinity Preparation of Sialylsulfo-glycopeptides from Hen Ovomucin and Their Characteristic Analysis

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a. Sugar chains were released from hen ovomucin by alkaline borohydride reaction. Neutral and acidic sugar chains were isolated by ion-exchange chromatography. Neutral sugar chains were purified Bio-Gel P-4 gel filtration. Acidic sugar chains were purified by ion-exchange and high performance liquid chromatographies. The chemical structures of sugar chains were analyzed by methylation and nuclear magnetic resonance analysis.b. Three novel neutral sugar chains (mono-, di- and trisaccharide) and three novel acidic sugar chains (tetra-, penta- and hexasaccharide) were finally clarified.c. Ovomucin was digested by actinase E.Neutral and acidic glycopeptides (sialyl- and sialylsulfo-glycopeptides) were separated by lysozyme-affinity chromatography which was developed in our previous paper.d. A new assay for evaluation of neutralizing activity against cholera toxin by using polystirene beads. This system was more high sensitive compared to microplate method.e. Sialyl- and sialylsulfo-glycopeptide isolated from hen ovomucin by lysozyme-affinity chromatography showed high neutralizing activity against cholera toxin.