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Anal Chem. 2005 Jul 1;77(13):4081-90.

Combining lectin microcolumns with high-resolution separation techniques for enrichment of glycoproteins and glycopeptides.

Madera M, Mechref Y, Novotny MV.

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Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.

Abstract

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.

PMID:: 15987113; [PubMed - indexed for MEDLINE]
PMCID:: PMC1472620

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