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

Milan Madera, Yehia Mechref, Milos V. Novotny

Research output: Contribution to journalArticlepeer-review

120 Scopus citations

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.

Original languageEnglish
Pages (from-to)4081-4090
Number of pages10
JournalAnalytical Chemistry
Volume77
Issue number13
DOIs
StatePublished - Jul 1 2005

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