METHODS

• US 20120071337A1
• Filed: 01/26/2010
• Published: 03/22/2012
• Est. Priority Date: 01/26/2009
• Status: Abandoned Application

• Current Assignee: Electrophoretics Limited (Proteome Sciences, PLC), Kings College London (University of London)
• Original Assignee: Electrophoretics Limited (Proteome Sciences, PLC), Kings College London (University of London)
• Inventors: Lovestone, Simon Harold, Güntert, Andreas Christian, Campbell, James, Byers, Helen Louise, O'Brien, Darragh P.W.
• Application Number:
  US13/146,121
• Publication Number:
  US 20120071337A1
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: G01N 2800/2821 Alzheimer
• CPC Class Codes: G01N 33/6896 Neurological disorders, e.g. Alzheimer's disease

Abstract

The invention provides a method for aiding the diagnosis or prognostic monitoring of Alzheimer'"'"'s disease in a subject, said method comprising; providing a sample of blood obtained from said patient; assaying the amount of gelsolin present in said sample; comparing the amount of gelsolin present in said sample to a reference amount of gelsolin present in a sample from a healthy subject, wherein detection of a gelsolin level in the sample from said patient which is lower than the gelsolin level in the reference sample indicates an increased likelihood of Alzheimer'"'"'s disease in said patient. Other markers are C1 protease inhibitor and ceruloplasmin. Both blood samples and tissue samples have been investigated.

Claims

1. A method for aiding the diagnosis of Alzheimer'"'"'s disease in a subject, said method comprising;

providing a sample of a blood derivative, wherein said blood derivative is serum or plasma, obtained from said subject;

assaying the amount of gelsolin present in said sample;

comparing the amount of gelsolin present in said sample to a reference amount of gelsolin present in a sample from a healthy subject, wherein detection of a gelsolin level in the sample from said subject which is lower than the gelsolin level in the reference sample indicates an increased likelihood of Alzheimer'"'"'s disease in said subject.

2. A method according to claim 1 wherein said sample comprises blood plasma.

3. A method according to claim 2 wherein said blood plasma has been depleted for one or more of albumin;

transferrin;

IgG;

IgA;

antitrypsin or haptoglobin.

4. A method according to claim 3, wherein said blood plasma has been depleted for each of albumin;

transferrin;

IgG;

IgA;

antitrypsin;

or haptoglobin.

5. A method according to claim 1 wherein the protein is detected by western blotting.

6. A method according to claim 1 wherein the protein is detected by bead suspension array.

7. A method according to claim 1 wherein the protein is detected by planar array.

8. A method according to claim 1 wherein the protein is detected by isobaric protein tagging or by isotopic protein tagging.

9. A method according to claim 1 wherein the protein is detected by mass spectrometer-based assay.

10. A method according to claim 1 wherein the protein is gelsolin and is detected by reference to one or more of the following peptides of Table B:

SEQ ID NO;

38, SEQ ID NO;

31, SEQ ID NO;

32.

11. A method for aiding the diagnosis or prognostic monitoring of Alzheimer'"'"'s disease in a subject, said method comprising;

providing a sample of a relevant tissue from said subject;

measuring the amount of one or more proteins selected from Gelsolin, C1 protease inhibitor and ceruloplasmin;

comparing the amount of said one or more proteins present in said sample to a reference amount of the same proteins in a sample from a healthy subject, wherein detection of a level different to that found in a reference sample indicates an increased likelihood of Alzheimer'"'"'s disease being present or developing or advancing in said subject.

12. A method for aiding the diagnosis or prognostic monitoring of Alzheimer'"'"'s disease in a subject, said method comprising;

(i) providing a sample of a relevant tissue from said subject;

(ii) measuring the amount of gelsolin; and

(iii) measuring the amount of one or more proteins selected from

C1 protease inhibitor;

ceruloplasmin;

clusterin;

complement c3;

serum amyloid P component;

alpha-2-macroglobulin;

gamma-fibrinogen;

complement factor H;

or

apolipoprotein E; and

(iv) comparing the amounts of said gelsolin and said one or more proteins present in said sample to a reference amount of the same proteins in a sample from a healthy subject, wherein detection of a level different to that found in a reference sample indicates an increased likelihood of Alzheimer'"'"'s disease being present or developing or advancing in said subject.

13. A method according to claim 12 wherein step (iii) comprises measuring the amount of one or mere proteins selected from:

clusterin;

complement c3;

serum amyloid P component;

alpha-2-macroglobulin;

gamma-fibrinogen;

complement factor H;

or

apolipoprotein E.

14. A method according to claim 12 wherein step (iii) comprises measuring the amount of one or more proteins selected from:

C1 protease inhibitor;

or

ceruloplasmin.

15. A method according to claim 14 comprising assaying the levels of each of gelsolin, C1 protease inhibitor and ceruloplasmin in a sample of blood from said subject.

16-19. -19. (canceled)

20. An assay device for use in the diagnosis of Alzheimer'"'"'s disease, which comprises a solid substrate having a location containing a material, which recognizes, binds to, or has affinity for a polypeptide, or a fragment, variant or mutant thereof, wherein the polypeptide is selected from gelsolin (SEQ ID NO:

1), C1 protease inhibitor (SEQ ID NO;

2), or Ceruloplasmin (SEQ ID NO;

3).

21. An assay device according to claim 20 in which the solid substrate has a plurality of locations each respectively containing a material which recognizes, binds to or has affinity for a polypeptide, or a fragment, variant or mutant thereof, wherein the polypeptide is selected from gelsolin (SEQ ID NO:

1), C1 protease inhibitor (SEQ ID NO;

2), or Ceruloplasmin (SEQ ID NO;

3).

22. An assay device according to claim 20, in which the material is an antibody or antibody chip.

23. An assay device according to claim 22, which has a unique addressable location for each antibody, thereby to permit an assay readout for each individual polypeptide or for any combination of polypeptides.

24. An assay device according to claim 21, including an antibody to a polypeptide wherein the polypeptide is selected from gelsolin (SEQ ID NO:

1), C1 protease inhibitor (SEQ ID NO;

2), or Ceruloplasmin (SEQ ID NO;

3).

25. An assay device according to claim 20, further having a location containing a material which recognizes, binds to or has affinity for glutathione S transferase P.

26. An assay device according to claim 25, in which the material is an antibody or antibody chip.

27. A kit for use in the diagnosis of Alzheimer'"'"'s disease, comprising an assay device according to claim 20, and means for detecting the amount of one or more of the polypeptides in a sample of body fluid taken from a subject.

28. A kit for use in the detection of gelsolin polypeptide, said kit comprising one or more of the following peptides of Table B:

SEQ ID NO;

30, SEQ ID NO;

31, SEQ ID NO;

32.

37. The kit according to claim 28 wherein one or more of the peptides comprises a heavy isotope.

38. The kit according to claim 28 wherein one or more of the peptides comprises a TMT tag.

39. The kit according to claim 37 comprising a further isotopic TMT tag for labeling of a sample polypeptide.

29. A kit for use in the diagnosis of Alzheimer'"'"'s disease, comprising one or more of the following peptides of Table B:

SEQ ID NO;

30, SEQ ID NO;

31, SEQ ID NO;

32.

30. A kit according to claim 29 comprising at least one further peptide of Table B.

40. The kit according to claim 29 wherein one or more of the peptides comprises a heavy isotope.

41. The kit according to claim 29 wherein one or more of the peptides comprises a TMT tag.

42. The kit according to claim 40 comprising a further isotopic TMT tag for labeling of a sample polypeptide.

43. The kit according to claim 30 wherein one or more of the peptides comprises a heavy isotope.

44. The kit according to claim 30 wherein one or more of the peptides comprises a TMT tag.

45. The kit according to claim 43 comprising a further isotopic TMT tag for labeling of a sample polypeptide.

31-33. -33. (canceled)

34. A method of determining the APOE ε

4 genotype of a subject, said method comprising assaying the C1 protease inhibitor level in a sample of blood from said subject.

35. A method of predicting the age of onset of Alzheimer'"'"'s disease for a subject, said method comprising assaying the ceruloplasmin levels in a sample of blood from said subject.

36. (canceled)