Binding molecule binding to traumatic brain injury-associated tau and methods of use thereof
The present invention provides detection reagents and method for determining risk of traumatic brain injury (TBI) and/or susceptibility to neurodegenerative disease in a subject.
1 . A binding molecule, wherein the binding molecule preferentially recognizes human traumatic brain injury (TBI)-associated tau relative to tau from a healthy normal control and comprises an amino acid sequence at least 96% identical to: (a) an antibody heavy chain variable domain and light chain variable domain of SEQ ID NO:42, or (b) a designed ankyrin repeat protein (DARPin) of SEQ ID NO:44, SEQ ID NO:48, SEQ ID NO:52, or SEQ ID NO:54; wherein differences from the heavy chain variable domain and light chain variable domain of (a) or the DARPIN of (b) are conservative substitutions.
2 . The binding molecule of claim 1 , wherein the binding molecule comprises the antibody heavy chain variable domain and light chain variable domain of SEQ ID NO:42.
3 . The binding molecule of claim 2 , wherein the binding molecule is a chimeric antibody, a single-chain antibody, an Fab, or an scFv.
4 . A nucleic acid that encodes the binding molecule of claim 1 .
5 . A vector comprising the nucleic acid of claim 4 .
6 . A phage comprising the vector of claim 5 .
7 . A phage comprising a nucleic acid encoding the binding molecule of claim 1 .
8 . A method of detecting traumatic brain injury (TBI)-associated tau in a subject, the method comprising: (a) incubating a sample from the subject with the binding molecule of claim 1 ; and (b) detecting an increased amount of TBI-associated tau bound to the binding molecule in the sample from the subject relative to the amount of TBI-associated tau bound to the binding molecule in a healthy normal control sample.
9 . The method of claim 8 , wherein the binding molecule comprises the antibody heavy chain variable domain and light chain variable domain of SEQ ID NO:42.
10 . The method of claim 9 , wherein the binding molecule is a chimeric antibody, a single-chain antibody, an Fab, or an scFv.
11 . A method for detecting traumatic brain injury (TBI) in a subject, the method comprising: (a) detecting TBI-associated tau in a sample from the subject obtained after head trauma or injury; and (b) comparing the level of TBI-associated tau protein in the sample from the subject with the level of TBI-associated tau protein in a sample from a healthy normal control; wherein detecting TBI-associated tau comprises incubating the sample from the subject with the binding molecule of claim 1 ; and wherein an elevated level of TBI-associated tau protein in the sample from the subject compared to the level of TBI-associated tau protein in the sample from the healthy normal control indicates TBI.
12 . The method of claim 11 , wherein the binding molecule comprises the antibody heavy chain variable domain and light chain variable domain of SEQ ID NO:42.
13 . The method of claim 12 , wherein the binding molecule is a chimeric antibody, a single-chain antibody, an Fab, or an scFv.
14 . The method of claim 11 , wherein the sample from the subject and the sample from the healthy normal control are blood, serum or cerebrospinal fluid (CSF).