Methods and compositions for the generation and use of conformation-specific antibodies

Patent metadata

Field	Value
Publication number	US-9688747-B2
Application number	US-201414776924-A
Country	US
Kind code	B2
Filing date	Mar 14, 2014
Priority date	Mar 15, 2013
Publication date	Jun 27, 2017
Grant date	Jun 27, 2017

Abstract

The present invention features methods and compositions for the generation and use of conformation-specific anti-bodies or fragments thereof.

First claim

The invention claimed is:

1. An isolated conformation-specific antibody or antibody fragment thereof, wherein said antibody or antibody fragment comprises:
(a) a heavy chain variable region comprising the heavy chain CDRs of SEQ ID NOs:1-3, and a light chain variable region comprising the light chain CDRs of SEQ ID NOs:4-6; or
(b) a heavy chain variable region comprising the heavy chain CDRs of SEQ ID NOs:7-9, and a light chain variable region comprising the light chain CDRs of SEQ ID NOs:10-12; and
wherein said isolated antibody or antibody fragment thereof binds specifically to the cis conformation of phosphorylated-Threonine231-tau protein (pT231-tau).

2. The isolated antibody or antibody fragment thereof of claim 1 , wherein the antibody or antibody fragment is a single chain antibody.

3. A pharmaceutical composition comprising the antibody or antibody fragment thereof of claim 1 and a pharmaceutically acceptable carrier.

4. A method of treating a tauopathy, traumatic brain injury (TBI), or stroke, said method comprising administering to a subject in need thereof the antibody or antibody fragment of claim 1 , in an amount sufficient to treat said tauopathy, TBI, or stroke.

5. The method of claim 4 , wherein said subject is predisposed to or is at an early stage of said tauopathy.

6. The method of claim 5 , further comprising determining the levels of CSF t-tau, pT181-tau, Aβ42, or ApoE4 in the subject.

7. The method of claim 5 , wherein said subject is predisposed by a history of repeated brain trauma.

8. A method for monitoring a therapeutic response in a subject treated with the antibody or antibody fragment of claim 1 , said method comprising measuring the level of cis pT231-tau or cis:trans pT231-tau ratio in a sample obtained from said subject, wherein a decrease in the level of cis pT231-tau or cis:trans pT231-tau ratio is indicative of an effective therapeutic response to said antibody or antibody fragment.

9. The method of claim 8 , further comprising measuring the levels of one or more of CSF t-tau, pT181-tau, Aβ42, or ApoE4.

10. A method of diagnosing a subject as having a tauopathy or having a predisposition to a tauopathy, said method comprising:
a. measuring the level of cis pT231-tau or cis:trans pT231-tau ratio in a sample obtained from said subject,
b. comparing said level of cis pT231-tau or cis:trans pT231-tau ratio in said sample with a non-diseased reference sample, wherein an elevated level of cis pT231-tau or an increase in cis:trans pT231-tau ratio as compared to said non-diseased reference sample indicates that said subject has or is predisposed to have said tauopathy; and
c. administering to said subject the antibody or antibody fragment of claim 1 in an amount sufficient to treat said tauopathy.

11. An isolated conformation-specific antibody or antibody fragment thereof, wherein said antibody or antibody fragment comprises:
a heavy chain variable region comprising the heavy chain CDRs of SEQ ID NOs:13-15, and a light chain variable region comprising the light chain CDRs of SEQ ID NOs:16-18,
and wherein said isolated antibody or antibody fragment thereof binds specifically to the trans conformation of phosphorylated-Threonine231-tau protein (pT231-tau).

12. The isolated antibody or antibody fragment thereof of claim 11 , wherein the antibody or antibody fragment thereof is a single chain antibody.

13. A pharmaceutical composition comprising the antibody or antibody fragment thereof of claim 11 and a pharmaceutically acceptable carrier.

14. A kit for diagnosing a subject as having a tauopathy or having a predisposition to a tauopathy comprising:
(a) a first isolated conformation-specific antibody or antibody fragment thereof comprising:
(i) a heavy chain variable region comprising the heavy chain CDRs of SEQ ID NOs:1-3, and a light chain variable region comprising the light chain CDRs of SEQ ID NOs:4-6; or
(ii) a heavy chain variable region comprising the heavy chain CDRs of SEQ ID NOs:7-9, and a light chain variable region comprising the light chain CDRs of SEQ ID NOs:10-12,
wherein said first antibody or antibody fragment thereof binds specifically to the cis conformation of pT231-tau,

(b) a second isolated conformation-specific antibody or antibody fragment thereof comprising:
a heavy chain variable region comprising the heavy chain CDRs of SEQ ID NOs:13-15, and a light chain variable region comprising the light chain CDRs of SEQ ID NOs:16-18,
wherein said second antibody or antibody fragment thereof binds specifically to the trans conformation of pT231-tau, and

(c)instructions for the use of the first antibody or antibody fragment thereof and the second antibody or antibody fragment thereof for diagnosing said subject as having a tauopathy or having a predisposition to said tauopathy.

Assignees

• Beth Israel Deaconess Medical Ct Inc

Inventors

• Lu Kun Ping
• Zhou Xiao Zhen

Classifications

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• A61P9/00

  Drugs for disorders of the cardiovascular system · CPC title

• A61P25/28

  for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia · CPC title

• C07K2317/20

  characterized by taxonomic origin · CPC title

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title