IDENTIFICATION OF SMALL MOLECULES RECOGNIZED BY ANTIBODIES IN SUBJECTS WITH NEURODEGENERATIVE DISEASES

• US 20100303805A1
• Filed: 06/01/2010
• Published: 12/02/2010
• Est. Priority Date: 06/02/2009
• Status: Active Grant

• Current Assignee: Opko Health, Inc., Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Moola, Muralidhar Reddy, Connell, Steven, Wilson, Johnnie, German, Dwight, Wilson, Rosemary, Kodadek, Thomas
• Granted Patent: US 9,551,721 B2
• Time in Patent Office: Days
• US Class Current: 424/130.1
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: G01N 2800/2821 Alzheimer
• CPC Class Codes: G01N 2800/2835 Movement disorders, e.g. Parkinson, Huntington, Tourette
• 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 33/6845 Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The present invention provides for the identification of individuals having neurodegenerative diseases (ND). Peptoids recognized by Parkinson'"'"'s Disease- and Alzheimer'"'"'s Disease-specific antibodies are identified, allowing one to diagnose or predict ND in subjects.

Claims

1. A method of detecting antibodies in an antibody-containing sample comprising:

(a) contacting an antibody-containing sample with a support having affixed thereto a peptoid having;

(i) the formula (1A);

2. The method of claim 1, further comprising obtaining said sample from a subject.

3. The method of claim 1, further comprising making a diagnosis of Alzheimer'"'"'s Disease for a subject from which said sample was obtained if antibody binding to said peptoid is greater than that observed for control non-diseased patients.

4. The method of claim 3, wherein one or more peptoid is selected from group consisting of

5. The method of claim 1, wherein said sample is contacted with more than one peptoid of claim 1.

6. The method of claim 5, wherein said sample is contacted with three structurally distinct peptoids.

7. The methods of claim 6, wherein the peptoids are AD1, AD2 and AD3.

8. The method of claim 1, wherein said support is a bead, a plate, a dipstick, a filter, a membrane a pin, or a well.

9. The method of claim 1, wherein said sample is blood, serum, saliva or CSF.

10. The method of claim 1, wherein detecting comprises RIA, FIA, ELISA, Western blot, flow cytometry, FRET, or surface plasmon resonance.

42. An antibody isolated by the steps comprising:

(a) contacing a sample containing an antibody indicative of a neurodegenerative disease with an immunoaffinity substrate comprising a peptoid of claim 1 and/or 11 that bind antibodies indicative of a neurodegenerative disease;

(b) eluting the bound antibodies from the immunoaffinity substrate.

11. A method of detecting antibodies in an antibody-containing sample comprising:

(a) contacting an antibody-containing sample with a support having affixed thereto a peptoid having;

(i) the formula (2A);

12. The method of claim 11, wherein, R²⁶, R²⁹, R³¹ and/or R³² are an alkylamine or N-butylamine.

13. The method of claim 11, wherein, R³⁰ is an alkenyl or an allyl group.

14. The method of claim 11, wherein, R³⁵ is an allyl group.

15. The method of claim 11, wherein, R³⁷ an alkoxy group

16. The method of claim 15, wherein the alkoxy group is an ethoxy group.

17. The method of claim 11, further comprising obtaining said sample from a subject.

18. The method of claim 11, further comprising making a diagnosis of Parkinson'"'"'s Disease for a subject from which said sample was obtained if antibody binding to said peptoid is greater than that observed for control non-diseased patients.

19. The method of claim 18, wherein the peptoid is selected from the group consisting of:

20. The method of claim 11, wherein said sample is contacted with more than one peptoid of formula 2A or 2B.

21. The method of claim 20, wherein said sample is contacted with three structurally distinct peptoids.

22. The methods of claim 21, wherein the peptoids are PD1, PD2 and PD3.

23. The method of claim 11, wherein said support is a bead, a plate, a dipstick, a filter, a membrane a pin, or a well.

24. The method of claim 11, wherein said sample is blood, serum, saliva or CSF.

25. The method of claim 11, wherein detecting comprises RIA, FIA, ELISA, Western blot, flow cytometry, FRET, or surface plasmon resonance.

26. A method of treating a subject suspected of having a neurodegenerative disease (ND) comprising:

(a) contacting an antibody-containing sample from said subject with one or more supports having affixed thereto a peptoid comprising;

(i) the formula (1A);

27. The method of claim 26, further comprising obtaining said sample from a subject.

28. The method of claim 26, further comprising making a diagnosis of Parkinson'"'"'s Disease for a subject from which said sample was obtained if antibody binding to the PD peptoid is greater than that observed for control non-diseased patients.

29. The method of claim 26, further comprising making a diagnosis of Alzheimers'"'"'s Disease for a subject from which said sample was obtained if antibody binding to the AD peptoid is greater than that observed for control non-diseased patients.

30. The method of claim 26, wherein said sample is contacted with more than one peptoid of formula (1A-D) and/or (2A-B).

31. The method of claim 30, wherein said sample is contacted with three structurally distinct peptoids that reactive with antibodies for each of PD and AD.

32. The method of claim 26, wherein said support is a bead, a plate, a dipstick, a filter, a membrane a pin, or a well.

33. The method of claim 26, wherein said sample is blood, serum, saliva or CSF.

34. The method of claim 26, wherein detecting comprises RIA, FIA, ELISA, Western blot, flow cytometry, FRET, or surface plasmon resonance.

35. The method of claim 26, further comprising making a treatment decision for said subject.

36. A kit for detecting antibodies that are indicative of a neurodegenerative disease, the kit comprising one or more peptoids comprising:

(i) the formula (1A);

37. The kit of claim 36, wherein the peptoid is coupled to a substrate.

38. The kit of claim 36, wherein one or more peptoids are selected from group consisting of:

39. A method of diagnosing AD in a subject suspected of having alzheimer'"'"'s disease comprising:

(a) exposing a serum sample from said patient to a microarray comprising a plurality of high-affinity peptoids having;

(i) the formula (1A);

40. An isolated antibody composition comprising an antibody isolated from the serum of a subject having a neurodegenerative disease, wherein the antibody specifically binds a peptoid comprising the following structure:

41. A kit for detecting antigens associated with Alzheimer'"'"'s disease or Parkison'"'"'s disease comprising the isolated antibody of claim 40.

43. A composition comprising a peptoid having the following formula:

(a) the formula (1A);

44. The composition of claim 43, further comprising an antibody indicative of a neurodegenerative disease.

45. A peptoid composition comprising the following formula:

46. The composition of claim 45, further comprising an antibody indicative of a neurodegenerative disease.

47. An isolated peptoid having the following formula:

(a) the formula (1A);

48. The isolated peptoid of claim 47, wherein the peptoid has a structure of:

49. The isolated peptoid of claim 47, wherein the peptoid has a structure of:

50. The isolated peptoid of claim 47, wherein the peptoid has a structure of:

51. The isolated peptoid of claim 47, wherein the peptoid has a structure of:

52. The isolated peptoid of claim 47, wherein the peptoid has a structure of:

53. The isolated peptoid of claim 47, wherein the peptoid has a structure of:

54. A method of detecting an antibody in a biological fluid comprising:

(a) incubating a peptoid array with a blocking buffer comprising a bacterial lysate at a concentration of 0.5 to 2 mg/ml;

(b) removing the blocking buffer;

(c) incubating the peptoid array with a sample solution comprising bacterial lysate; and

(d) detecting antibody binding to the peptoid array.