Sugar-coated microarrays: a novel slide surface for the high-throughput analysis of glycans.
The development of DNA and protein microarrays represents a significant advance in transcriptomics and proteomics research. Such arrays allow the high-throughput, parallel analysis of protein occurrence and interactions and gene expression. However, this advance has not been matched by equivalent technology for analysis of glycomes. One reason for this is that compared to proteins, it is difficult to reliably immobilise populations of chemically and structurally diverse glycans. We describe the development of a new microarray slide surface to which diverse glycan structures can be directly immobilised without prior derivatisation of the slide surface or any modification of the arrayed samples. The slides can be used to produce comprehensive microarrays of carbohydrates, glycoproteins and proteoglycans using isolated samples or cell extracts. Using standard microarray equipment, a series of carbohydrate microarrays were generated and probed with a panel of monoclonal antibodies with specificities for glycan epitopes. The arrays were highly reproducible, stable, and could be stored dry for several months. Glycans play central roles in development, carcinogenesis, cell adhesion, and immunity and are increasingly the subject of therapeutic approaches. We anticipate that the development of carbohydrate microarrays will be important for the high-throughput analysis of glycans and their molecular interactions.
Willats
William G T
WG
Centre for Plant Sciences, University of Leeds, U.K. w.g.t.willats@leeds.ac.uk
Rasmussen
Svend Erik
SE
Kristensen
Tina
T
Mikkelsen
Jørn Dalgaard
JD
Knox
J Paul
JP
D000818:Animals; D000911:Antibodies, Monoclonal; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D013499:Surface Properties
D000911:Antibodies, Monoclonal; D011134:Polysaccharides; D011137:Polystyrenes