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.
Madera
Milan
M
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
Mechref
Yehia
Y
Novotny
Milos V
MV
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