Glycobiology Advance Access originally published online on November 24, 2003
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Glycobiology vol 14 no 2 pp. 197-203, 2004
© Oxford University Press 2004; all rights reserved.
Intact cell adhesion to glycan microarrays
3 Department of Pharmacology and Molecular Sciences, The Johns Hopkins School of Medicine, 725 N. Wolfe Street, Baltimore, MD 21205; 4 Instituto de Microbiologia Prof. Paulo de Goes, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil; and 5 Glycominds, Ltd., Lod 71291, Israel
Received on September 9, 2003; revised on October 22, 2003; accepted on October 22, 2003
A rapid and reproducible method was developed to detect and quantify carbohydrate-mediated cell adhesion to glycans arrayed on glass slides. Monosaccharides and oligosaccharides were covalently attached to glass slides in 1.7-mm-diameter spots (200 spots/slide) separated by a Teflon gasket. Primary chicken hepatocytes, which constitutively express a C-type lectin that binds to nonreducing terminal N-acetylglucosamine residues, were labeled with a fluorescent dye and incubated in 1.3-µL aliquots on the glycosylated spots. After incubating to allow cell adhesion, nonadherent cells were removed by immersing the slide in phosphate buffered saline, inverting, and centrifuging in a sealed custom acrylic chamber so that cells on the derivatized spots were subjected to a uniform and controlled centrifugal detachment force while avoiding an air–liquid interface. After centrifugation, adherent cells were fixed in place and detected by fluorescent imaging. Chicken hepatocytes bound to nonreducing terminal GlcNAc residues in different linkages and orientations but not to nonreducing terminal galactose or N-acetylgalactosamine residues. Addition of soluble GlcNAc (but not Gal) prior to incubation reduced cell adhesion to background levels. Extension of the method to CD4+ human T-cells on a 45-glycan diversity array revealed specific adhesion to the sialyl Lewis x structure. The described method is a robust approach to quantify selective cell adhesion using a wide variety of glycans and may contribute to the repertoire of tools for the study of glycomics.
1 These authors contributed equally to this work.
2 To whom correspondence should be addressed; e-mail: schnaar{at}jhu.edu
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  H. Tateno, N. Uchiyama, A. Kuno, A. Togayachi, T. Sato, H. Narimatsu, and J. Hirabayashi A novel strategy for mammalian cell surface glycome profiling using lectin microarray Glycobiology, October 1, 2007; 17(10): 1138 - 1146. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N Dotan, R. Altstock, M Schwarz, and A Dukler Anti-Glycan Antibodies as Biomarkers for Diagnosis and Prognosis Lupus, July 1, 2006; 15(7): 442 - 450. [Abstract] [PDF]
|
|
|
|
|
|
S. Angeloni, J.L. Ridet, N. Kusy, H. Gao, F. Crevoisier, S. Guinchard, S. Kochhar, H. Sigrist, and N. Sprenger Glycoprofiling with micro-arrays of glycoconjugates and lectins Glycobiology, January 1, 2005; 15(1): 31 - 41. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Blixt, S. Head, T. Mondala, C. Scanlan, M. E. Huflejt, R. Alvarez, M. C. Bryan, F. Fazio, D. Calarese, J. Stevens, et al. Printed covalent glycan array for ligand profiling of diverse glycan binding proteins PNAS, December 7, 2004; 101(49): 17033 - 17038. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. V. Su, P. Hong, S. Baik, O. A. Negrete, K. B. Gurney, and B. Lee DC-SIGN Binds to HIV-1 Glycoprotein 120 in a Distinct but Overlapping Fashion Compared with ICAM-2 and ICAM-3 J. Biol. Chem., April 30, 2004; 279(18): 19122 - 19132. [Abstract] [Full Text] [PDF]
|
|