Glycoprofiling with micro-arrays of glycoconjugates and lectins
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Author
Angeloni, Silvia
Ridet, J.L.
Kusy, N.
Gao, Hui
Sigrist, Hans
Sprenger, Nobert
Abstract
To facilitate deciphering the information content in the
glycome, thin film-coated photoactivatable surfaces were applied
for covalent immobilization of glycans, glycoconjugates, or
lectins in microarray formats. Light-induced immobilization
of a series of bacterial exopolysaccharides on photoactivatable
dextran-coated analytical platforms allowed covalent
binding of the exopolysaccharides. Their specific galactose
decoration was detected with fluorescence-labeled lectins.
Similarly, glycoconjugates were covalently immobilized and
displayed glycans were profiled for fucose, sialic acid,
galactose, and lactosamine epitopes. The applicability of
such platforms for glycan profiling was further tested with
extracts of Caco2 epithelial cells. Following spontaneous
differentiation or on pretreatment with sialyllactose, Caco2
cells showed a reduction of specific glycan epitopes. The
changed glycosylation phenotypes coincided with altered
enteropathogenic E. coli adhesion to the cells. This microarray
strategy was also suitable for the immobilization of lectins
through biotin-neutravidin-biotin bridging on platforms
functionalized with a biotin derivatized photoactivatable
dextran. All immobilized glycans were specifically and
differentially detected either on glycoconjugate or lectin
arrays. The results demonstrate the feasibility and versatility
of the novel platforms for glycan profiling.
Publication Reference
Glycobiology 15(1):31-41
Year
2004-09-01