US 2003/0113768 A1 · Synthetic substrate for high specificity enzymatic assays

Abstract

Novel synthetic enzyme substrates, enhanced to have improved enzymatic specificity, are disclosed. These synthetic enzyme substrates consist of a substrate peptide that has had its specificity further improved by additional synthetic moieties, selected by combinatorial chemistry techniques, that act to sterically block non-target enzymes. These “steric restrictor” moieties may be labeled to produce a detectable signal upon enzymatic reaction. These novel substrates are particularly useful for improved enzyme substrate microarrays. Specific applications for improved protease substrate microarrays are discussed. A variety of applications for these improved protease substrate microarrays are also disclosed, including proteomics research, protease discovery, protease binding site characterization, diagnosis of the protease composition of biological samples, monitoring the angiogenic status of a tumor, monitoring the status of arthritis and other inflammatory diseases, and the discovery and optimization of novel drugs that modify or inhibit protease activity.

Claims22

Dependent claims

A synthetic enzyme substrate, comprising:

a peptide substrate moiety capable of independently serving as a substrate for a target enzyme,
one or more steric blocking groups covalently coupled to the peptide substrate moiety, wherein said steric blocking groups are polymers comprised of two or more non-identical monomeric units,
and wherein the sequence of the steric blocking groups is selected based upon the blocking groups ability to protect the peptide substrate from serving as a substrate for non-target enzymes, while preserving the ability of the peptide substrate to serve as a substrate for the target enzyme.

An assay device for enzymatic activity, employing one or more synthetic substrates bound to a solid support, in which one or more of these synthetic substrates are capable of serving as a substrate for enzymatic activity for a target enzyme, and in which one or more of the synthetic substrates comprise;

a peptide substrate moiety capable of independently serving as a substrate for a target enzyme;
one or more steric blocking groups covalently coupled to the peptide substrate moiety;
wherein said steric blocking groups are polymers comprised of two or more non-identical monomeric units,
and wherein the sequence of the steric blocking groups are selected based upon their ability to protect the peptide substrate from serving as a substrate for non-target enzymes, while preserving the ability of the peptide substrate to serve as a substrate for a target enzyme.

1 more element

Dependencies are inferred from the claim text.

Synthetic substrate for high specificity enzymatic assays

Life of this patent

Synthetic substrate for high specificity enzymatic assays

Life of this patent

Abstract

Claims22