Printed covalent glycan array for ligand profiling of diverse glycan binding proteins

doi:10.1073/pnas.0407902101

Printed covalent glycan array for ligand profiling of diverse glycan binding proteins

^aGlycan Synthesis and Protein Expression Core-D, Consortium for Functional Glycomics, ^dDNA Microarray Core Facility, and Departments of ^bMolecular Biology, ^hChemistry, and ^eImmunology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037; ^fSidney Kimmel Cancer Center, 10835 Altman Row, San Diego, CA 92121; ^gDepartment of Biochemistry and Molecular Biology, University of Oklahoma Health Science Center, 975 NE 10th Street, BRC 411B, P.O. Box 26901, Oklahoma City, OK 73104; ⁱMedical Research Center National Institute for Medical Research, Mill Hill, London NW7, United Kingdom; ^jDepartment of Molecular Cell Biology and Immunology, Vrije Universiteit Medical Centre, 1007 MB, Amsterdam, The Netherlands; and^kShemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya ul, Moscow, 117997, V-437, Russian Federation

Contributed by Chi-Huey Wong, October 26, 2004

Abstract

Here we describe a glycan microarray constructed by using standard robotic microarray printing technology to couple amine functionalized glycans to an amino-reactive glass slide. The array comprises 200 synthetic and natural glycan sequences representing major glycan structures of glycoproteins and glycolipids. The array has remarkable utility for profiling the specificity of a diverse range of glycan binding proteins, including C-type lectins, siglecs, galectins, anticarbohydrate antibodies, lectins from plants and microbes, and intact viruses.

Footnotes

↵ c To whom correspondence should be addressed. E-mail: olablixt{at}scripps.edu.
Author contributions: O.B., S.H., and T.M. designed research; O.B., C.S., M.E.H., and R.A. performed research; O.B., M.E.H., M.C.B., F.F., D.C., J.S., N.R., D.J.S., J.J.S., I.v.D., D.R.B., I.A.W., R.C., N.B., and C.-H.W. contributed new reagents/analytic tools; O.B. and C.S. analyzed data; O.B. and J.C.P. wrote the paper; and J.C.P. was the grant holder.
Abbreviations: CVN, cyanovirin-N; DC-SIGN, dendritic cell-specific intercellular adhesion molecule-1-grabbing nonintegrin; ECA, Erythrina cristagalli; GBP, glycan binding protein; NHS, N-hydroxysuccinimide.
↵ l Compound library was produced by the Consortium for Functional Glycomics (www.functionalglycomics.org).
↵ m Symbol structure insets represent the principal glycan structures recognized. See Fig. 2 for a complete list of structures.
↵ n The array using the ELISA format uses biotinylated glycans adsorbed to streptavidin. It is currently available for use by investigators through the Consortium for Functional Glycomics (www.functionalglycomics.org) and soon will be replaced by the printed microarray described here.
Freely available online through the PNAS open access option.