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Quantitative analysis of carbohydrate-protein interactions using glycan microarrays: Determination of surface and solution dissociation constants
Journal article   Peer reviewed

Quantitative analysis of carbohydrate-protein interactions using glycan microarrays: Determination of surface and solution dissociation constants

Pi-Hui Liang, Sheng-Kai Wang and Chi-Huey Wong
Journal of the American Chemical Society, Vol.129(36), pp.11177-11184
12/09/2007

Abstract

Carbohydrate-protein interactions on surface and in solution were quantitatively measured by a glycan microarray. Assessing carbohydrate affinities is typically difficult due to weak affinities and limited sources of structurally complex glycans. We described here a sensitive, high-throughput, and convenient glycan microarray technology for the simultaneous determination of a wide variety of parameters in a single experiment using small amounts of materials. Assay systems based on this technology were developed to analyze multivalent interactions and determine the surface dissociation constant (K D,surf ) for surface-coated mannose derivatives with mannose binding lectins and antibodies. Competition experiments that employed monovalent ligands in solution yielded K D and K i values in solution similar to equilibrium binding constants obtained in titration microcalorimetry and surface plasmon resonance experiments. © 2007 American Chemical Society.

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