Combining lectin microcolumns with high-resolution separation techniques for enrichment of glycoproteins and glycopeptides.
Silica-based lectin microcolumns are described in this study together with the chemical procedures necessary for their preparation. The analytical merits of Canavalia ensiformis and Sambucus nigra lectins, [immobilized on activated macroporous silica], such as binding capacity, trapping reproducibility, and substrate selectivity, have been evaluated using model glycoproteins. The described microcolumns are applicable to high-pressure analytical schemes utilizing microvalving procedures, washing steps, and quantitative desorption for LC/MS analysis. The described analytical systems are amenable to the applications aiming at fractionation of complex glycopeptide mixtures and determination of the sites of glycosylation.
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
Milan
M
Madera
Yehia
Y
Mechref
Milos V
MV
Novotny
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry*; D054788:Ribosome Inactivating Proteins
D006020:Glycopeptides; D006023:Glycoproteins; D037121:Plant Lectins; C050600:Sambucus nigra lectins; C039268:lectin, Canavalia; D054788:Ribosome Inactivating Proteins