Quantitative analysis of carbohydrate-protein interactions using glycan microarrays: determination of surface and solution dissociation constants
AssigneeAcademia Sinica
InventorsChi-Huey Wong, Pi-Hui Liang
Life of this patent
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Abstract
A method, system and device to identify, study and/or mimic carbohydrate-protein interactions on cell surfaces and in solution measured by a glycan microarray. In some instances the method, system and device uses very small quantities of carbohydrate as low as attomol. In some instances the system, method and device is high-throughput. The small quantity sensitivity may allow for close placement of carbohydrate array members wherein due to close proximity multivalent interactions with proteins may be identified.
Claims13
A dense carbohydrate microarray for quantitative protein measurements, comprising:
- a solid substrate configured to provide a supporting substance;
- and
- a reactive layer fabricated on the solid substrate, the reactive layer comprising at least one reactive group of small molecules;
- wherein the at least one reactive group of small molecules is configured to bind with at least one protein, wherein the at least one protein are immobilized on the solid substrate via the reactive layer;
- wherein the carbohydrate microarray is configured to perform a quantitative measurement of binding ability with the at least one protein;
- wherein at least three different concentrations of the at least one protein are provided in one chip;
- wherein the binding affinity between the small molecules and the at least one protein can be measured;
- wherein the small molecules comprise at least one of carbohydrates and glycolipids;
- wherein the small molecules are disposed relative to each other to allow the at least one protein to multivalently bond with the small molecules.
A kit comprising:
- a high throughput carbohydrate microarray comprising carbohydrate spots on a solid substrate, each of the carbohydrate spots having a reactive group of at least one of carbohydrates and glycolipids, wherein the carbohydrates spots are configured to react with and bind to at least one protein;
- wherein the reactive groups are disposed relative to each other to allow the at least one protein to multivalently bond with the reactive groups;
- a microarray reader;
- and
- instructions for use.
A method of identifying a protein bound to a microarray of attomol quantity carbohydrates, the method comprising:
- forming a glycan microarray of spotted sugars in concentrations each of about 10−18 mole;
- adding a labeled protein to the microarray;
- incubating the microarray;
- and,
- using an array reader to identify labeled proteins on the glycan microarray.
Dependencies are inferred from the claim text.