US 2006/0216721 A1 · Compositions and methods for detecting protease activity

Abstract

The invention provides a method of determining activity of a protease. The method can include the steps of (a) providing a protease substrate including a protein moiety attached to a nucleic acid moiety and a ligand moiety; (b) contacting the protease substrate with a protease under conditions wherein the protease catalyzes cleavage of the protein moiety, thereby producing a proteolytic product wherein the nucleic acid moiety is separated from at least a portion of the protein moiety and the ligand moiety; (c) contacting the proteolytic product with a receptor under conditions wherein the ligand moiety binds to the receptor to form a complex; (d) separating the complex from the nucleic acid moiety, thereby forming a separation product including the nucleic acid moiety; (e) contacting the separation product with a probe nucleic acid under conditions wherein the nucleic acid moiety hybridizes to a complementary sequence of the probe; and (f) detecting hybridization of the separation product to the probe, thereby determining activity of the protease.

Claims32

Dependent claims

A method of determining activity of a protease, comprising:

(a) providing a protease substrate comprising a protein moiety attached to a nucleic acid moiety and a ligand moiety;
(b) contacting said protease substrate with a protease under conditions wherein said protease catalyzes cleavage of said protein moiety, thereby producing a proteolytic product wherein said nucleic acid moiety is separated from at least a portion of said protein moiety and said ligand moiety;
(c) contacting said proteolytic product with a receptor under conditions wherein said ligand moiety binds to said receptor to form a complex;
(d) separating said complex from said nucleic acid moiety, wherein said receptor is attached to a solid support, thereby forming a separation product comprising said nucleic acid moiety;
(e) contacting said separation product with a probe nucleic acid under conditions wherein said nucleic acid moiety hybridizes to a complementary sequence of said probe;
and
(f) detecting hybridization of said separation product to said probe, thereby determining activity of said protease.

4 more elements

Dependencies are inferred from the claim text.

Compositions and methods for detecting protease activity

Life of this patent

Compositions and methods for detecting protease activity

Life of this patent

Abstract

Claims32