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PMID: 15987113 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

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

Analytical chemistry ·Vol. 77 ·No. 13 ·2005-07-01 ·Pages 4081-90

Madera M, Mechref Y, Novotny MV

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.

MeSH Terms
Amino Acid Sequence Chromatography, Affinity/methods Glycopeptides/chemistry Glycoproteins/chemistry Plant Lectins/chemistry Ribosome Inactivating Proteins
Chemicals
Glycopeptides Glycoproteins Plant Lectins Sambucus nigra lectins lectin, Canavalia Ribosome Inactivating Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Madera Milan
Department of Chemistry, Indiana University, Bloomington, Indiana 47405, USA.
Mechref Yehia
Novotny Milos V
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Article Info
Journal
Analytical chemistry
Abbr.
Anal Chem
ISSN
0003-2700
DOI
10.1021/ac050222l
Published
2005-07-01
Pages
4081-90
Language
English
Region
United States
NLM ID
0370536
PMCID
PMC1472620
Subset
IM
Grants
NIGMS NIH HHS · R01 GM024349 · United States
NIGMS NIH HHS · GM24349 · United States
External Links
PubMed source DOI: 10.1021/ac050222l PMC Full Text View journal details
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