Antibody based reagents that specifically recognize toxic oligomeric forms of tau

Abstract

The invention relates to antibodies, antibody fragments and binding agents that specifically recognize oligomeric tau but do not bind to monomelic tau, fibrillar tau or non-disease associated forms of tau.

1 . An antibody or antibody fragment that specifically recognizes oligomeric tau but does not bind monomeric tau, fibrillar tau or non-disease associated forms of tau. 
     
     
         2 . The antibody or antibody fragment of  claim 1 , wherein the oligomeric tau is dimeric tau or trimeric tau. 
     
     
         3 . The antibody or antibody fragment of  claim 2 , wherein the oligomeric tau is trimeric tau. 
     
     
         4 . The antibody or antibody fragment of  claim 1 , wherein the oligomeric tau is soluble. 
     
     
         5 . The antibody or antibody fragment of  claim 1 , wherein said antibody fragment is isolated according to a method comprising the steps of:
 a. a negative panning of a scFV phage library wherein said negative panning eliminates phage that bind to non-desired antigens wherein said negative panning comprises serially contacting phage with:
 (i) a generic protein; and 
 (ii) mononeric forms of tau; 
   and monitoring the binding of said phage to the generic protein and monomeric forms of tau using Atomic Force Microscope (AFM) Imaging and repeating steps (i) and (ii) until no phage is observed binding to antigen by said AFM imaging to produce an aliquot of phage;   b. contacting the aliquot of phage with tau oligomers and incubating for time sufficient to allow binding of phage to said oligomers; and   c. eluting the bound phage particles from step (b).   
     
     
         6 . An antibody or antibody fragment isolated according to a method comprising the steps of:
 (a) negative panning a scFV phage library comprising serially contacting phage with:
 (i) a generic protein; and 
 (ii) mononeric forms of tau; 
 and until less than 5% of the phage is observed binding to antigen, which produces an aliquot of phage; 
   (b) positive panning of the aliquot from step (a) comprising contacting the aliquot of phage from step (a) with tau oligomers, and incubating for time sufficient to allow binding of phage to said brain derived tau oligomers; and   (c) eluting the bound phage particles from step (b).   
     
     
         7 . The antibody or antibody fragment of  claim 5 , wherein the tau oligomer is trimeric tau 4N1R. 
     
     
         8 . The antibody or antibody fragment of  claim 5 , wherein the generic protein is bovine serum albumin (BSA). 
     
     
         9 . The antibody or antibody fragment of  claim 5 , wherein the negative panning further comprises serially contacting phage with brain derived control samples that do not contain oligomeric tau. 
     
     
         10 . The antibody or antibody fragment of  claim 6 , wherein the observing of the binding of the phage to the antigen is by using Atomic Force Microscope (AFM) Imaging. 
     
     
         11 . The antibody or antibody fragment of  claim 6 , wherein the negative panning is repeated until less than 0-10% phage was observed by AFM imaging as binding to antigen in step (a). 
     
     
         12 . The antibody fragment of  claim 1 , wherein said antibody fragment does not contain the constant domain region of an antibody. 
     
     
         13 . An antibody fragment comprising amino acid sequence SEQ ID NO:1, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, or SEQ ID NO:19. 
     
     
         14 . The antibody fragment of  claim 13 , wherein the antibody fragment comprises amino acid sequence SEQ ID NO:1 , SEQ ID NO:9 or SEQ ID NO:11. 
     
     
         15 . A binding molecule that binds to oligomeric tau and does not bind monomelic tau, fibrillar tau or non-disease associated forms of tau, wherein the binding molecule comprises the sequence of SEQ ID NO:1, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, or SEQ ID NO:19. 
     
     
         16 . A method of inhibiting the aggregation of tau comprising contacting a composition that comprises tau oligomers with an antibody, antibody fragment or binding molecule of  claim 1 . 
     
     
         17 . The method of  claim 16 , wherein said aggregation of tau is in a cell. 
     
     
         18 . The method of  claim 16 , wherein said aggregation of tau is in brain tissue. 
     
     
         19 . The method of  claim 16 , wherein said contacting with an antibody, antibody fragment or binding molecule decreases the rate of formation of tau aggregates as compared to said rate in the absence of composition or binding molecule. 
     
     
         20 . A method of detecting the presence of tau in a physiological sample comprising contacting a sample with an antibody, antibody fragment or a binding molecule of  claim 1  and determining the binding of said composition with said tissue sample wherein binding of said composition to said tissue sample is indicative of the presence of tau oligomers in said tissue sample wherein said presence of said tau oligomers is indicative of early stage AD, frontotemporal dementia, other tauopathies or neurodegeneration following traumatic brain injury. 
     
     
         21 . The method of  claim 20 , wherein the physiological sample is brain tissue, serum, cerebrospinal fluid (CSF), blood, urine or saliva. 
     
     
         22 . The method of  claim 16 , wherein the tau oligomers are dimeric trimeric tau. 
     
     
         23 . The method of  claim 22 , wherein the tau oligomers are trimeric tau. 
     
     
         24 . The method of  claim 22 , wherein the tau oligomers are soluble. 
     
     
         25 . A method of preventing or inhibiting the accumulation of tau in the brain of a mammal comprising administering to said mammal a composition comprising an antibody fragment or a binding molecule of  claim 1 . 
     
     
         26 . The method of  claim 20 , wherein the physiological sample is spinal tissue.