Lectin purification on affinity columns containing reductively aminated disaccharides.
A method is described for isolating lectins in pure form and quantitative yield in a single step by affinity chromatography on aminoethyl polyacrylamide gels containing reductively aminated disaccharide residues. The affinity columns were prepared in two steps: (a) direct reductive amination of the disaccharide and aminoethyl gel with sodium cyanoborohydride in aqueous solution at pH 9; (b) N-acetylation of excess amino groups. Affinity columns prepared by reductive amination of lactose, melibiose, maltose, and di-N-acetylchitobiose were used to purify the following lectins: lactose, peanut, castor bean; melibiose, Bandeiraea simplicifolia; maltose, jack bean, common lentil; di-N-acetylchitobiose, wheat germ. These columns are extremely stable, have good flow rates, and high binding capacities.
D002846:Chromatography, Affinity / Q000379:methods; D004187:Disaccharides; D004912:Erythrocytes / Q000276:immunology; D006386:Hemagglutination Tests; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007785:Lactose; D037102:Lectins / Q000302:isolation & purification*; D011485:Protein Binding; D013045:Species Specificity
D004187:Disaccharides; D037102:Lectins; D007785:Lactose