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Glycobiology Advance Access published online on September 1, 2004
Glycobiology, doi:10.1093/glycob/cwh143
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Received May 26, 2004
Revised July 20, 2004
Accepted August 24, 2004

ORIGINAL ARTICLES

Glycoprofiling with micro-arrays of glycoconjugates and lectins

S. Angeloni 1, J. L. Ridet 2, N. Kusy 2, H. Gao 1, F. Crevoisier 1, S. Guinchard 1, S. Kochhar 2, H. Sigrist 1, N. Sprenger 2*

1 CSEM S.A., Centre Suisse d'Electronique et de Microtechnique, Rue Jaquet-Droz 1, CH-2000 Neuchâtel, Switzerland
2 Nestlé Research Center, Nestec Ltd., Vers-chez-les-Blanc, CH-1000 Lausanne 26, Switzerland

* To whom correspondence should be addressed. E-mail: norbert.sprenger{at}rdls.nestle.com.


  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. Ligth-induced immobilization of a series of bacterial exopolysaccharides on photoactivateable dextran (OptoDex®) coated analytical platforms (PhotoChips) 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 upon 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 photoactivable dextran (OptoDex-Biotin). All immobilized glycans were specifically and differentially detected either on glycoconjugate- or on lectin arrays. The results demonstrate the feasibility and versatility of the novel platforms for glycan profiling.

Keywords: glycoarray; glycosylation; lectins; exopolysaccharides; glycomics.
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