Monoclonal Antibody And Use Thereof

Abstract

An antibody capable of recognizing amyloid β while not recognizing amyloid β precursor proteins, and a method for using the same.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a drawing showing the specificity of the 1A10 antibody by western blotting.

FIG. 2 is a drawing showing the specificity of the 1C3 antibody by immunological precipitation.

FIG. 3 is a drawing showing the specificity of the 82E1 antibody by western blotting (A: 82E1 antibody, B; 6E10 antibody).

FIG. 4 is a drawing showing the specificity of the 82E1 antibody by western blotting (A: 82E1 antibody, B; 6E10 antibody).

FIG. 5 shows the results of standard curves obtained by assaying a synthetic amyloid β using an ELISA kit prepared in Example 10, wherein A shows the result of assaying a synthetic amyloid P (1-40) using a combination of the 1A10 antibody and HRP labeled 82E1 antibody and B is a result of assaying a synthetic amyloid β (1-42) using a combination of the 1C3 antibody and HRP labeled 82E1 antibody.

FIG. 6 is a drawing showing the results of immunological staining of the left cerebral hemisphere, including the cortical layer and hippocampus (A: mouse to which the 82E1 antibody was administered, B: control mouse).

FIG. 7 is a drawing showing the results of immunological staining of the left cerebral hemisphere (basal nuclei) (A: mouse to which the 82E1 antibody was administered, B: control mouse).

FIG. 8 is a drawing showing deposition of amyloid β (1-40) and amyloid β (1-42) in segments, in terms of the proportion (%) of the immunological staining positive region in the entire region.

FIG. 9 is a drawing showing the results of ELISA assay of amyloid β (1-42) and amyloid β (x-42) in the brain.

Claims

1. A monoclonal antibody which can recognize an N-terminus peptide of an amyloid β, but not an amyloid β precursor protein.

2. The monoclonal antibody according to claim 1, wherein the N-terminus peptide of the amyloid β is a peptide shown by the following amino acid sequence (a):

3. The monoclonal antibody according to claim 1, wherein the N-terminus peptide of the amyloid β is a peptide shown by the following amino acid sequence (b):

4. The monoclonal antibody according to claim 1, which is obtained by immunizing an animal with a bound substance of the N-terminus peptide of the amyloid β and a biological high molecular compound as a first antigen, immunizing the thus-immunized animal with a bound substance of another N-terminus peptide of the amyloid β, which is comparatively shorter than the peptide used for the first antigen, and a biological high molecular compound as a second antigen, and collecting the antibody from the animal.

5. The monoclonal antibody according to claim 1, which is obtained by immunizing an animal with a bound substance of a peptide shown by the amino acid sequence (a) and a biological high molecular compound as a first antigen, immunizing the thus-immunized animal with a bound substance of a peptide shown by the amino acid sequence (b) and a biological high molecular compound as a second antigen, and collecting the antibody from the animal: (a) DAEFRHDSGYEVHHQK (Sequence ID No. 1) and(b) DAEFR (Sequence ID No. 2).

6. The monoclonal antibody according to any one of claims 1 to 5, wherein the monoclonal antibody is a chimeric antibody.

7. The monoclonal antibody according to any one of claims 1 to 5, wherein the monoclonal antibody is a humanized antibody.

8. A kit for assaying amyloids β comprising a first reagent containing an antibody which can recognize the N-terminus peptide of amyloid β, but not amyloid β precursor proteins, and a second reagent containing an antibody which can recognize amyloid β (1-40) or amyloid β (1-42).

9. The kit for assaying amyloid β according to claim 8, wherein the antibody recognizing an amyloid β (1-40) or amyloid β (1-42) is an antibody recognizing the C-terminus peptide of the amyloid β.

10. The kit for assaying amyloid β according to claim 8, wherein the C-terminus peptide of amyloid β is a peptide shown by the following amino acid sequence (c):

11. The kit for assaying amyloid β according to claim 10, wherein the kit is for assaying amyloid β (1-40).

12. The kit for assaying amyloid β according to claim 8, wherein the C-terminus peptide of amyloid β is a peptide shown by the following amino acid sequence (d):

13. The kit for assaying amyloid β according to claim 12, wherein the kit is for assaying amyloid β (1-42).

14. A method for assaying amyloid β comprising causing an antibody which can recognize the N-terminus peptide of an amyloid β, but not amyloid β precursor proteins, and an antibody which can recognize amyloid β (1-40) or amyloid β (1-42) to react with an amyloid β in a sample to be assayed.

15. The method for assaying amyloid β according to claim 14, wherein the antibody recognizing an amyloid β (1-40) or amyloid β (1-42) is an antibody recognizing the C-terminus peptide of the amyloid β.

16. A method for preparing a monoclonal antibody of claim 1 comprising immunizing an animal with a bound substance of the N-terminus peptide of the amyloid β and a biological high molecular compound as a first antigen, immunizing the thus-immunized animal with a bound substance of another N-terminus peptide of the amyloid β, which is comparatively shorter than the peptide used for the first antigen, and a biological high molecular compound as a second antigen, and collecting the antibody from the animal.

17. A therapeutic agent for Alzheimer's disease comprising the monoclonal antibody according to any one of claims 1 to 7 as an active ingredient.

18. A deposition inhibitor of amyloid β (1-40) or amyloid β (1-42) comprising the monoclonal antibody according to any one of claims 1 to 7 as an active ingredient.

19. A method for treatment of Alzheimer's disease comprising administering the monoclonal antibody according to any one of claims 1 to 7.

20. A method for inhibiting deposition of amyloid β (1-40) or amyloid β (1-42) comprising administering the monoclonal antibody according to any one of claims 1 to 7.