Assays for measuring nucleic acid binding proteins and enzyme activities

• US 20080254494A1
• Filed: 10/31/2007
• Published: 10/16/2008
• Est. Priority Date: 09/17/1998
• Status: Abandoned Application

• Current Assignee: BioVeris Corporation (Roche Holding AG)
• Original Assignee: BioVeris Corporation (Roche Holding AG)
• Inventors: Kenten, John H., Kibbey, Maura C., Heroux, Jeffrey A.
• Application Number:
  US11/981,753
• Publication Number:
  US 20080254494A1
• Time in Patent Office: Days
• US Class Current: 435/23
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 2521/301 Endonuclease
• CPC Class Codes: C12Q 2522/101 Single or double stranded nucleic acid binding proteins

Abstract

Processes for measuring DNA or RNA binding proteins, specific nucleic acids, as well as enzyme activities using labeled nucleic acids of labeled protein/peptide molecules are provided.

Claims

1-2. -2. (canceled)

3. A method for assaying a sample for the presence of an enzyme activity that cleaves peptides or proteins, the method comprising:

a) mixing at least one predetermined peptide or protein containing at least one electrochemiluminescent label with a sample which may contain a peptide or protein-cleaving enzyme;

b) incubating the mixture of step (a) under conditions which allow cleavage of said peptide or protein; and

c) analyzing the incubated mixture for the presence of cleaved peptide or protein.

4. The method of claim 3, wherein the electrochemiluminescent label comprises ruthenium, osmium or rhenium.

5. The method of claim 3, wherein the electrochemiluminescent label comprises a bipyridyl or phenanthrolyl-containing complex of ruthenium.

6. The method of claim 3, wherein the electrochemiluminescent label comprises RuBpy.

7. The method of claim 3, wherein analyzing the incubated mixture comprises measuring the amount of cleaved peptide or protein in the incubated mixture.

8. The method of claim 3, wherein the peptide- or protein-cleaving enzyme is a protease.

9. The method of claim 3, wherein the peptide- or protein-cleaving enzyme is a peptidase.

10. The method of claim 3, wherein the electrochemiluminescent label is linked to the protein or peptide by a covalent bond, or a specific interaction with a binding group.

11. The method of claim 3, wherein analyzing the incubated mixture comprises analyzing an incubated mixture, which includes an ECL coreactant.

12. The method of claim 11, wherein the ECL coreactant comprises a tertiary amine.

13. The method of claim 3, wherein the peptide or protein has a capture moiety attached thereto.

14. The method of claim 13, wherein the capture moiety is selected from the group consisting of antibody, antigen, receptor, ligand, and a nucleic acid base pairing polymer.

15. A method of detecting an inhibitor of a peptide- or protein-cleaving enzyme, comprising:

a) combining to form a mixture i) at least one predetermined peptide or protein containing at least one electrochemiluminescent label, ii) at least one predetermined peptide- or protein-cleaving enzyme, and iii) a sample, wherein the sample may contain an inhibitor of the peptide- or protein-cleaving enzyme;

b) incubating the mixture of step (a) under conditions which allow cleavage of said peptide or protein; and

c) analyzing the incubated mixture for the presence of cleaved peptide or protein.

16. The method of claim 15, wherein the electrochemiluminescent label is linked to the protein or peptide by a covalent bond, or a specific interaction with a binding group.

17. The method of claim 15, wherein the electrochemiluminescent label comprises a bipyridyl or phenanthrolyl-containing complex of ruthenium.

18. The method of claim 15, wherein analyzing the incubated mixture comprises measuring the amount of cleaved peptide or protein in the incubated mixture.

19. The method of claim 15, wherein the peptide- or protein-cleaving enzyme is a protease.

20. The method of claim 15, wherein the peptide- or protein-cleaving enzyme is a peptidase.