Dissolved Protein Arthritis Markers

• US 20120003665A1
• Filed: 12/14/2009
• Published: 01/05/2012
• Est. Priority Date: 05/09/2006
• Status: Active Grant

• Current Assignee: University of British Columbia
• Original Assignee: University of British Columbia
• Inventors: GHAHARY, AZIZ, MAKSYMOWYCH, WOLODYMYR WALTER PETER, KILANI, RUHANGIZ TAGHI
• Granted Patent: US 9,791,458 B2
• Time in Patent Office: Days
• US Class Current: 435/7.4
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: G01N 2800/102 Arthritis; Rheumatoid arthritis, i.e. inflammation of peripheral joints
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere

Abstract

Methods and kits for diagnosing arthritis are provided. The methods may involve detection of 14-3-3 eta or gamma proteins in a sera or synovial fluid sample.

Claims

1-13. -13. (canceled)

14. A method for evaluating an arthritic condition or disease state in a mammalian test subject, comprising:

obtaining a test biological sample from said test subject; and

detecting at least one 14-3-3 protein isoform in said test sample, wherein said 14-3-3 protein isoform is selected from the group consisting of a 14-3-3 gamma isoform and a 14-3-3 eta isoform.

15. The method according to claim 14, wherein said detecting step comprises an immunological-based technique.

16. The method according to claim 15, wherein said immunological-based technique comprises combining said test sample with antibodies specific for said 14-3-3 protein isoform.

17. The method according to claim 16, wherein said antibodies are specific for the 14-3-3 eta isoform.

18. The method according to claim 16, wherein said antibodies are specific for the 14-3-3 gamma isoform.

19. The method according to claim 14, wherein said detecting step comprises mass spectroscopy.

20. The method according to claim 14, wherein an increased level of said 14-3-3 protein isoform in said test sample in comparison with a control level of said 14-3-3 protein isoform in a suitable control sample is a diagnostic indicator of said arthritic condition or disease state in the test subject.

21. The method according to claim 14, wherein a test level of said 14-3-3 protein isoform in said test sample, compared with a control level of said 14-3-3 protein isoform in a suitable control sample, is a diagnostic indicator of said arthritic condition or disease state in the test subject.

22. The method according to claim 14, wherein a decreased level of said 14-3-3 protein isoform in said test sample in comparison with a previously-determined level of said 14-3-3 protein isoform in a previous test sample taken from said test subject is indicative of efficacy of an ongoing therapeutic regimen for the test subject.

23. The method according to claim 14, wherein said 14-3-3 gamma isoform comprises an amino acid sequence of SEQ ID NO:

2.

24. The method according to claim 14, wherein said 14-3-3 eta isoform comprises an amino acid sequence of SEQ ID NO:

3.

25. The method according to claim 14, further comprising detection of at least one additional arthritis marker in said test sample useful for detecting and/or diagnosing an arthritic condition or disease state in the test subject.

26. The method according to claim 25, wherein said at least one additional arthritis marker is a matrix metalloproteinase.

27. The method according to claim 14, wherein said arthritic condition or disease state is rheumatoid arthritis.

28. The method according to claim 14, wherein said test sample is selected from the group consisting of blood, serum, plasma, and synovial fluid.

29. A method for characterization of arthritis, the method comprising:

measuring the expression of at least one 14-3-3 protein isoform in a biological test sample from an individual suspected of having arthritis or an elevated risk of arthritis, wherein increased expression of said at least one 14-3-3 protein isoform in said test sample relative to expression of said 14-3-3 protein isoform in a suitable control sample is a positive indicator of arthritis or elevated risk of arthritis in said individual.

30. The method according to claim 29, wherein the at least one 14-3-3 protein isoform is detected with antibodies that specifically bind to said 14-3-3-protein isoform.

31. The method according to claim 30, wherein the antibodies are added to said test sample, and incubated for a period of time sufficient to allow antibody binding to a 14-3-3-protein epitope.

32. A kit for evaluating/characterizing an arthritic condition in a mammalian subject, comprising at least one reagent that specifically binds to at least one 14-3-3 protein isoform selected from the group consisting of a 14-3-3 gamma isoform and a 14-3-3 eta isoform, together with instructions for using the kit for diagnostic or prognostic purposes.

33. A kit according to claim 32, comprising a first reagent that specifically binds to a 14-3-3 gamma isoform, and a second reagent that specifically binds to a 14-3-3 eta isoform.

34. A kit according to claim 32, wherein said reagent is selected from the group consisting of antibodies, organic chemicals, inorganic chemicals, and combinations thereof.

35. A kit according to claim 32, additionally comprising at least one additional reagent that specifically binds to a matrix metalloproteinase.