Methods of diagnosing ALS
The invention relates to an epitope protection assay for use in diagnosis, prognosis and therapeutic intervention in diseases, for example, involving polypeptide aggregation, such as prion infections. The methods of the invention first block accessible polypeptide target epitope with a blocking agent. After denaturation of the polypeptide, a detecting agent is used to detect protein with target epitope that was inaccessible during contact with the blocking agent. The invention also relates to novel amyotrophic lateral sclerosis-specific epitopes and their uses to make antibodies, and to the novel antibodies and uses thereof.
1. A method of detecting or diagnosing amyotrophic lateral sclerosis (ALS) in a subject comprising the steps of:
(a) contacting a test sample of said subject with an antibody that specifically binds DLGKGGNEESTKTGNAGS (SEQ ID NO:1) and/or NPLSRKHGGPKDEE (SEQ ID NO: 2) to produce an antibody-antigen complex;
(b) measuring the amount of the antibody-antigen complex in the test sample; and;
(c) detecting a difference in the amount of antibody-antigen complex in the test sample as compared to a control which is indicative of amyotrophic lateral sclerosis.
2. The method of claim 1 , wherein the antibody specifically binds DLGKGGNEESTKTGNAGS (SEQ ID NO:1).
3. The method according to claim 2 , wherein the antibody is specific for residues 3-12 of SEQ ID NO:1.
4. The method according to claim 2 , wherein the antibody is monoclonal, polyclonal, chimeric or humanized.
5. The method according to claim 2 , wherein the antibody is an antibody fragment.
6. The method according to claim 5 , wherein the antibody fragment is a Fab, Fab′, F(ab′) 2 , scFv, dsFv, ds-scFv, dimer, minibody, diabody, or multimer thereof.
7. The method according to claim 3 , wherein the antibody is monoclonal, polyclonal, chimeric or humanized.
8. The method according to claim 3 , wherein the antibody is an antibody fragment.
9. The method according to claim 8 , wherein the antibody fragment is a Fab, Fab′, F(ab′) 2 , scFv, dsFv, ds-scFv, dimer, minibody, diabody, or multimer thereof.
10. The method of claim 1 , wherein the antibody specifically binds NPLSRKHGGPKDEE (SEQ ID NO: 2).
11. The method according to claim 10 , wherein the antibody is monoclonal, polyclonal, chimeric or humanized.
12. The method according to claim 10 , wherein the antibody is an antibody fragment.
13. The method according to claim 12 , wherein the antibody fragment is a Fab, Fab′, F(ab′) 2 , scFv, dsFv, ds-scFv, dimer, minibody, diabody, or multimer thereof.
14. The method according to claim 1 , wherein the antibody is monoclonal, polyclonal, chimeric or humanized.
15. The method according to claim 1 , wherein the antibody is an antibody fragment.
16. The method according to claim 15 , wherein the antibody fragment is a Fab, Fab′, F(ab′) 2 , scFv, dsFv, ds-scFv, dimer, minibody, diabody, or multimer thereof.