期刊
BIOSENSORS & BIOELECTRONICS
卷 21, 期 8, 页码 1451-1458出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2005.06.008
关键词
carbohydrate microarrays; protein-carbohydrate interaction; oxime bonding; biochips; oligosaccharide
Carbohydrate-protein interactions play important biological roles in biological processes. But there is a lack of hi.-h-throughput methods to elucidate recognition events between carbohydrates and proteins. This paper reported a convenient and efficient method for preparing oligosaccharide microarrays, wherein the underivatized oligosaccharide probes were efficiently immobilized on aminooxyacetyl functionalized class surface by formation of oxime bonding with the carbonyl group at the reducing end of the suitable carbohydrates via irreversible condensation. Prototypes of carbohydrate microarrays containing 10 oligosaccharides were fabricated on aminooxyacetyl functionalized glass by robotic arrayer. Utilization of the prepared carbohydrate microarrays for the characterization of carbohydrate-protein interaction reveals that carbohydrates with different structural features selectively bound to the corresponding lectins with relative binding affinities that correlated with those obtained from solution-based assays. The limit of detection (LOD) for lectin ConA on the fabricated carbohydrate microarrays was determined to be similar to 0.008 mu g/mL. Inhibition experiment with soluble carbohydrates also demonstrated that the binding affinities of lectins to different carbohydrates could be analyzed quantitatively by determining IC50 values of the soluble carbohydrates with the carbohydrate microarrays. This work provides a simple procedure to prepare carbohydrate microarray for high-throughput parallel characterization of carbohydrate-protein interaction. (c) 2005 Elsevier B.V. All rights reserved.
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