Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research

Henriette Lodberg Pedersen, Jonatan Ulrik Fangel, Barry McCleary, Christian Ruzanski, Maja Gro Rydahl, Marie-Christine Ralet, Vladimir Farkas, Laura von Schantz, Susan E. Marcus, Mathias Christian Franch Andersen, Rob Field, Mats Ohlin, J. Paul Knox, Mads Hartvig Clausen, William George Tycho Willats

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Microarrays are powerful tools for high throughput analysis, and hundreds or thousands of molecular interactions can be assessed simultaneously using very small amounts of analytes. Nucleotide microarrays are well established in plant research, but carbohydrate microarrays are much less established, and one reason for this is a lack of suitable glycans with which to populate arrays. Polysaccharide microarrays are relatively easy to produce because of the ease of immobilizing large polymers noncovalently onto a variety of microarray surfaces, but they lack analytical resolution because polysaccharides often contain multiple distinct carbohydrate substructures. Microarrays of defined oligosaccharides potentially overcome this problem but are harder to produce because oligosaccharides usually require coupling prior to immobilization. We have assembled a library of well characterized plant oligosaccharides produced either by partial hydrolysis from polysaccharides or by de novo chemical synthesis. Once coupled to protein, these neoglycoconjugates are versatile reagents that can be printed as microarrays onto a variety of slide types and membranes. We show that these microarrays are suitable for the high throughput characterization of the recognition capabilities of monoclonal antibodies, carbohydrate-binding modules, and other oligosaccharide-binding proteins of biological significance and also that they have potential for the characterization of carbohydrate-active enzymes.
Original languageEnglish
JournalJournal of Biological Chemistry
Volume287
Issue number47
Pages (from-to)39429-39438
ISSN0021-9258
DOIs
Publication statusPublished - 2012

Keywords

  • Glycobiology
  • Glycoconjugate
  • Glycomics
  • Microarray
  • Plant Cell Wall
  • Carbohydrate-binding Modules
  • Oligosaccharides

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