Methods and compositions for glycosidase assays
AssigneeReddy US Therapeutics, Inc.
InventorsSivaram Pillarisetti, Uday Saxena, Dongyan Wang
Life of this patent
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Abstract
The present invention is directed to compositions and methods for measuring enzymatic activity, particularly glycosidase activity. Methods of the present invention include assays for quantitatively determining the amount of glycosidase activity in a sample. The present invention also provides methods for the diagnosis of metastatic and inflammatory processes in vitro and in vivo. The present invention further provides compositions and methods for high throughput assays for identifying compounds that effect glycosidase activity.
Claims20
A method for detecting glycosidase activity, comprising,
- a) mixing a sample suspected of containing glycosidase activity with a composition comprising a substrate-binding partner to form a reaction mixture;
- b) removing an aliquot of the reaction mixture to a solid substrate comprising a complementary binding partner to bind the substrate-binding partner;
- c) adding a labeled complementary binding partner;
- d) detecting the label;
- and
- e) determining the amount of glycosidase activity.
The method of claim wherein the glycosidase activity is due to the presence of an endoglycosidase.
A method for detecting compounds that alter a glycosidase, comprising,
- a) mixing a compound with a glycosidase;
- b) adding the mixture of a) with a composition comprising a substrate-binding partner to form a reaction mixture;
- c) removing an aliquot of the reaction mixture to a solid substrate comprising a complementary binding partner to bind the substrate-binding partner;
- d) adding a labeled complementary binding partner;
- e) detecting the label;
- and
- f) determining the change in glycosidase activity.
A method for detecting metastases, comprising,
- a) mixing a sample from one suspected of having metastases with a composition comprising a substrate-binding partner to form a reaction mixture, wherein the substrate is the substrate for a glycosidase;
- b) removing an aliquot of the reaction mixture to a solid substrate comprising a complementary binding partner to bind the substrate-binding partner;
- c) adding a labeled complementary binding partner;
- d) detecting the label;
- and
- e) determining the amount of glycosidase activity.
Dependencies are inferred from the claim text.