Antibodies specific for N-terminal truncated and pyroglutamate modified amyloid-beta peptides
The present invention pertains to novel 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. An isolated antibody comprising:
(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,
wherein,
the isolated antibody binds to an Aβ peptide or variant thereof.
2. The antibody 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 antibody according to claim 1 , wherein said antibody is a monoclonal antibody.
4. The antibody 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 antibody 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 antibody 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 antibody 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 antibody 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 antibody 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
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.
10. The antibody 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 antibody according to claim 1 , wherein said antibody is Aβ 6-1-6 (Deposit No. DSM ACC 2924).
12. The antibody according to claim 1 , wherein said antibody is Aβ 24-2-3 (Deposit No. DSM ACC 2926).
13. The antibody according to claim 1 , wherein said antibody is a humanized or chimeric antibody, or an antibody fragment.
14. The antibody according to claim 1 , wherein the antibody is a human antibody.
15. The antibody according to claim 1 , wherein the antibody is a diabody or a single chain antibody.
16. The antibody 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 antibody 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 antibody according to claim 1 , wherein the antibody is labeled.
19. The antibody according to claim 1 , wherein the antibody is immobilised on a solid phase.
20. The antibody 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. A composition comprising a hybridoma cell line selected from the group consisting of DSM ACC 2923; DSM ACC 2924; DSM ACC 2925; and DSM ACC 2926.
22. A composition comprising the antibody of claim 1 and, optionally, a pharmaceutically acceptable carrier.
23. A diagnostic kit comprising the antibody according to claim 1 and instructions for use, and, optionally, at least one additional biologically active substance.
24. The diagnostic kit according to claim 23 , wherein said at least one additional biologically active substance comprises an inhibitor of glutaminyl cyclase.
25. The antibody according to claim 16 , wherein x is an integer between 18 and 42.
26. The antibody according to claim 16 , wherein x is an integer between 30 and 42.
27. The antibody according to claim 17 , wherein x is an integer between 18 and 42.
28. The antibody according to claim 17 , wherein x is an integer between 30 and 42.