Pedersen, Henriette Lodberg4; Fangel, Jonatan Ulrik4; McCleary, Barry5; Ruzanski, Christian11; Rydahl, Maja Gro4; Ralet, Marie-Christine7; Farkas, Vladimir8; von Schantz, Laura12; Marcus, Susan E.10; Andersen, Mathias Christian Franch1; Field, Rob11; Ohlin, Mats12; Knox, J. Paul10; Clausen, Mads Hartvig1; Willats, William George Tycho4
1 Department of Chemistry, Technical University of Denmark2 Organic Chemistry, Department of Chemistry, Technical University of Denmark3 Center for Nanomedicine and Theranostics, Center, Technical University of Denmark4 University of Copenhagen5 Megazyme International Ireland Ltd.6 John Innes Centre7 National Institute for Agronomic Research8 Slovak Academy of Sciences9 Lund University10 University of Leeds11 John Innes Centre12 Lund University
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.
Journal of Biological Chemistry, 2012, Vol 287, Issue 47, p. 39429-39438