Diagnostic antibody assay
Diagnostic assays for the diagnosis of amyloidosis, in particular, Alzheimer's disease, and related aspects. In particular, monoclonal antibodies and an antibody assay are provided.
1. A method for detecting an Aβ peptide or variants thereof comprising:
contacting an antibody that binds to an Aβ peptide or variant thereof and a sample potentially comprising an Aβ peptide or variants thereof; and
detecting binding of the antibody and the Aβ peptide or variant thereof;
wherein the antibody comprises
(i) a light chain comprising a variable part, the variable part of the light chain of said antibody comprising a polypeptide
(a) encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 49, SEQ ID NO: 53, SEQ ID NO: 57, and SEQ ID NO: 61 or
(b) having an amino acid sequence selected from the group consisting of SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 58, and SEQ ID NO: 62; or
(ii) a heavy chain comprising a variable part; the variable part of the heavy chain of said antibody comprising a polypeptide
(a) encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 51, SEQ ID NO: 55, SEQ ID NO: 59, and SEQ ID NO: 63 or
(b) having an amino acid sequence selected from the group consisting of SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 60, and SEQ ID NO: 64.
2. The method according to claim 1 , wherein the antibody binds an Aβ peptide with a dissociation constant (KD) value of at least 10 −7 M.
3. The method according to claim 1 , wherein said antibody is a monoclonal antibody.
4. The method according to claim 1 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part; and
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 49, SEQ ID NO: 53, SEQ ID NO: 57, and SEQ ID NO: 61, or having an amino acid sequence selected from the group consisting of SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 58, and SEQ ID NO: 62.
5. The method according to claim 1 , wherein:
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence selected from SEQ ID NO: 51, SEQ ID NO: 55, SEQ ID NO: 59, and SEQ ID NO: 63, or having an amino acid sequence selected from SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 60, and SEQ ID NO: 64.
6. The method according to claim 1 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 49 or having an amino acid sequence of SEQ ID NO: 50;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 51, or having an amino acid sequence of SEQ ID NO: 52.
7. The method according to claim 1 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 53 or having an amino acid sequence of SEQ ID NO: 54;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 55, or having an amino acid sequence of SEQ ID NO: 56.
8. The method according to claim 1 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 57 or having an amino acid sequence of SEQ ID NO: 58;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 59, or having an amino acid sequence of SEQ ID NO: 60.
9. The method according to claim 1 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 61 or having an amino acid sequence of SEQ ID NO: 62;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 63, or having an amino acid sequence of SEQ ID NO: 64.
10. The method according to claim 1 , wherein said antibody is selected from the group consisting of:
Aβ 5-5-6
(Deposit No. DSM ACC 2923);
Aβ 6-1-6
(Deposit No. DSM ACC 2924);
Aβ 17-4-3
(Deposit No. DSM ACC 2925); and
Aβ 24-2-3
(Deposit No. DSM ACC 2926).
11. The method according to claim 1 , wherein said antibody is Aβ 6-1-6 (Deposit No. DSM ACC 2924).
12. The method according to claim 1 , wherein said antibody is Aβ 24-2-3 (Deposit No. DSM ACC 2926).
13. The method according to claim 1 , wherein said antibody is a chimeric antibody or an antibody fragment.
14. The method according to claim 1 , wherein the antibody is a human immunoglobulin other than the variable part of the light chain and the variable part of the heavy chain.
15. The method according to claim 1 , wherein the antibody is a diabody or a single chain antibody.
16. The method according to claim 1 , wherein the antibody binds to an epitope of an antigen selected from the group consisting of:
pGlu-Aβ 3-38 ;
pGlu-Aβ 3-40 ;
pGlu-Aβ 3-42 ; and
pGlu-Aβ 3-x variants,
wherein x is an integer between 10 and 42.
17. The method according to claim 1 , wherein the antibody comprises a complementarity determining region that binds to an antigen selected from the group consisting of:
pGlu-Aβ 3-38 ;
pGlu-Aβ 3-40 ;
pGlu-Aβ 3-42 ; and
pGlu-Aβ 3-x variants,
wherein x is an integer between 10 and 42.
18. The method according to claim 1 , wherein the antibody is labeled.
19. The method according to claim 1 , wherein the antibody is immobilised on a solid phase.
20. The method according to claim 1 , wherein the antibody is obtained from any one of hybridoma cell lines DSM ACC 2923, DSM ACC 2924, DSM ACC 2925, or DSM ACC 2926.
21. The method according to claim 16 , wherein x is an integer between 18 and 42.
22. The method according to claim 16 , wherein x is an integer between 30 and 42.
23. The method according to claim 17 , wherein x is an integer between 18 and 42.
24. The method according to claim 17 , wherein x is an integer between 30 and 42.
25. A method for in vitro diagnosis of an amyloid-associated disease or condition, comprising:
contacting an antibody that binds to an Aβ peptide or variant thereof of claim 1 and a sample from a subject suspected to be afflicted with said disease or condition, and
detecting binding of the antibody to a pGlu-amyloid protein, from the sample;
wherein the antibody comprises
(i) a light chain comprising a variable part, the variable part of the light chain of said antibody comprising a polypeptide
(a) encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 49, SEQ ID NO: 53, SEQ ID NO: 57, and SEQ ID NO: 61 or
(b) having an amino acid sequence selected from the group consisting of SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 58, and SEQ ID NO: 62; or
(ii) a heavy chain comprising a variable part; the variable part of the heavy chain of said antibody comprising a polypeptide
(a) encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 51, SEQ ID NO: 55, SEQ ID NO: 59, and SEQ ID NO: 63 or
(b) having an amino acid sequence selected from the group consisting of SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 60, and SEQ ID NO: 64.
26. The method according to claim 25 , wherein the amyloid-associated disease or condition is Alzheimer's disease.
27. The method according to claim 25 , wherein the pGlu-amyloid protein comprises pGlu-Aβ peptide.
28. The method according to claim 25 , wherein the antibody binds an Aβ peptide with a dissociation constant (KD) value of at least 10 −7 M.
29. The method according to claim 25 , wherein said antibody is a monoclonal antibody.
30. The method according to claim 25 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part; and
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence selected from the group consisting of SEQ ID NO: 49, SEQ ID NO: 53, SEQ ID NO: 57, and SEQ ID NO: 61, or having an amino acid sequence selected from the group consisting of SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 58, and SEQ ID NO: 62.
31. The method according to claim 25 , wherein:
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence selected from SEQ ID NO: 51, SEQ ID NO: 55, SEQ ID NO: 59, and SEQ ID NO: 63, or having an amino acid sequence selected from SEQ ID NO: 52, SEQ ID NO: 56, SEQ ID NO: 60, and SEQ ID NO: 64.
32. The method according to claim 25 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 49 or having an amino acid sequence of SEQ ID NO: 50;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 51, or having an amino acid sequence of SEQ ID NO: 52.
33. The method according to claim 25 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 53 or having an amino acid sequence of SEQ ID NO: 54;
the isolated antibody comprises a heavy chain;
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 55, or having an amino acid sequence of SEQ ID NO: 56.
34. The method according to claim 25 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 57 or having an amino acid sequence of SEQ ID NO: 58;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 59, or having an amino acid sequence of SEQ ID NO: 60.
35. The method according to claim 25 , wherein:
the isolated antibody comprises a light chain;
the light chain comprises a variable part;
the variable part of the light chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 61 or having an amino acid sequence of SEQ ID NO: 62;
the isolated antibody comprises a heavy chain;
the heavy chain comprises a variable part; and
the variable part of the heavy chain of said antibody comprises a polypeptide encoded by a nucleotide sequence of SEQ ID NO: 63, or having an amino acid sequence of SEQ ID NO: 64.
36. The method according to claim 25 , wherein said antibody is selected from the group consisting of:
Aβ 5-5-6
(Deposit No. DSM ACC 2923);
Aβ 6-1-6
(Deposit No. DSM ACC 2924);
Aβ 17-4-3
(Deposit No. DSM ACC 2925); and
Aβ 24-2-3
(Deposit No. DSM ACC 2926).
37. The method according to claim 25 , wherein said antibody is Aβ 6-1-6 (Deposit No. DSM ACC 2924).
38. The method according to claim 25 , wherein said antibody is Aβ 24-2-3 (Deposit No. DSM ACC 2926).
39. The method according to claim 25 , wherein said antibody is a chimeric antibody or an antibody fragment.
40. The method according to claim 25 , wherein the antibody is a human immunoglobulin other than the variable part of the light chain and the variable part of the heavy chain.
41. The method according to claim 25 , wherein the antibody is a diabody or a single chain antibody.
42. The method according to claim 25 , wherein the antibody binds to an epitope of an antigen selected from the group consisting of:
pGlu-Aβ 3-38 ;
pGlu-Aβ 3-40 ;
pGlu-Aβ 3-42 ; and
pGlu-Aβ 3-x variants,
wherein x is an integer between 10 and 42.
43. The method according to claim 25 wherein the antibody comprises a complementarity determining region that binds to an antigen selected from the group consisting of:
pGlu-Aβ 3-38 ;
pGlu-Aβ 3-40 ;
pGlu-Aβ 3-42 ; and
pGlu-Aβ 3-x variants,
wherein x is an integer between 10 and 42.
44. The method according to claim 25 , wherein the antibody is labeled.
45. The method according to claim 25 , wherein the antibody is immobilised on a solid phase.
46. The method according to claim 25 , wherein the antibody is obtained from any one of hybridoma cell lines DSM ACC 2923, DSM ACC 2924, DSM ACC 2925, or DSM ACC 2926.
47. The method according to claim 42 , wherein x is an integer between 18 and 42.
48. The method according to claim 42 , wherein x is an integer between 30 and 42.
49. The method according to claim 43 , wherein x is an integer between 18 and 42.
50. The method according to claim 43 , wherein x is an integer between 30 and 42.