Improved affinity chromatographic purification of D-mannose-N-acetyl-D-glucosamine-specific lectin from the bark of Sophora japonica eliminating the loss by sugar specific self-aggregation.
A novel D-mannose-N-acetyl-D-glucosamine-specific lectin of Sophora japonica bark, B-SJA-II, which showed self-aggregation based on sugar specificity, was purified by affinity chromatography on maltamyl-Sepharose subsequent to chromatographic separation on lactamyl-Sepharose to remove a major D-galactose-N-acetyl-D-galactosamine specific lectin, B-SJA-I. However, the yield of this method was low as a result of the sugar-specific precipitation and binding to other glycoproteins. A modified method was developed to circumvent this problem. All the purification procedures, except for the final chromatographic separation, were carried out in the presence of the haptenic sugar and the sugar-specific adsorption of B-SJA-II onto the adsorbent was carried out in a dialysis bag by gradually removing the sugar. This method gave a yield eight times higher than the original method.
Ueno
M
M
Department of Chemistry, Faculty of Science, Ochanomizu University, Tokyo, Japan.
Arai
T
T
Kojima
K
K
Ogawa
H K
HK
Matsumoto
I
I
Seno
N
N
D000107:Acetylation; D002241:Carbohydrates / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D003956:Dialysis; D004187:Disaccharides / Q000302:isolation & purification; D037102:Lectins / Q000302:isolation & purification*; D037121:Plant Lectins; D010944:Plants / Q000737:chemistry*
D002241:Carbohydrates; D004187:Disaccharides; D037102:Lectins; D037121:Plant Lectins; C035093:4-O-mannopyranosyl-2-acetamido-2-deoxyglucose