Saccharide microarrays for high-throughput interrogation of glycan-protein binding interactions.
This chapter describes two methods for fabricating microarrays of saccharides for display and interrogation with binding proteins, using fluorescence detection. The first approach is based on the rapid immobilization of heparan sulphate glycans upon commercially available aminosilane slides via their reducing ends. The second approach is based on the use of a hydrazide-derivatized self-assembled monolayer (SAM) on a gold-coated slide surface. Both provide for efficient and chemoselective attachment and anchoring of oligosaccharide probes via their reducing ends, enabling the large-scale arraying of natural saccharides without cumbersome pre-derivatization. The latter platform, in particular, also has the potential for use with other biophysical readout methods including matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy, surface plasmon resonance, and quartz crystal microbalances. These microarray platforms provide a facile approach for interrogating multiple carbohydrate-protein interactions in a high-throughput manner using minimal quantities of reagents. They provide an essential new experimental strategy in the growing armoury of the glycomics toolkit.
Powell
Andrew K
AK
School of Biological Sciences, University of Liverpool, Crown Street, L69 7ZB, Liverpool, UK.
Zhi
Zheng-Liang
ZL
Turnbull
Jeremy E
JE
D000818:Animals; D054794:Glycomics / Q000379:methods; D006801:Humans; D046228:Microarray Analysis / Q000295:instrumentation / Q000379:methods*; D008954:Models, Biological; D009844:Oligosaccharides / Q000032:analysis*; D011134:Polysaccharides / Q000378:metabolism*; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013499:Surface Properties
D009844:Oligosaccharides; D011134:Polysaccharides; D011506:Proteins