Methods for detecting traumatic brain injury

Abstract

The present invention provides detection reagents and method for determining risk of traumatic brain injury (TBI), assessment of the amount of neuronal damage, and/or susceptibility to neurodegenerative disease in a subject.

What is claimed is: 
     
         1 . A designed ankyrin repeat protein (DARPin) comprising
 (a) an N-Terminal Capping ankyrin repeat (AR) encoded by (SEQ ID NO:30),   (b) a C-Terminal Capping AR encoded by (SEQ ID NO:31), and   (c) three to six AR modules of about 30 to 35 amino acids, wherein each AR module binds with a target.   
     
     
         2 . The DARPin of  claim 1 , wherein all of the AR modules are the same. 
     
     
         3 . The DARPin of  claim 1 , wherein one or more of the AR modules differ. 
     
     
         4 . The DARPin of  claim 1 , wherein an AR module binds with TDP-43, tau, abeta or alpha-synuclein. 
     
     
         5 . A nucleic acid encoding the DARPin of  claim 1 . 
     
     
         6 . A vector comprising the nucleic acid of  claim 5 . 
     
     
         7 . The vector of  claim 6 , wherein the vector is a pIT2 vector. 
     
     
         8 . The vector of  claim 7 , wherein the pIT2 vector lacks a BsaI restriction site. 
     
     
         9 . The vector of  claim 7 , wherein the vector lacks a PelB signal and comprises a DsbA signal. 
     
     
         10 . A phage comprising the vector of  claim 6 . 
     
     
         11 . A method for determining risk of traumatic brain injury (TBI), assessment of the amount of neuronal damage, and/or susceptibility to neurodegenerative disease in a subject, comprising the steps of:
 (A) providing samples obtained from a subject at two or more times post-injury;   (B) assessing levels of toxic variants of TDP-43, tau, abeta and/or alpha-synuclein in the sample by detecting toxic variants of TDP-43, tau, abeta and/or alpha-synuclein protein levels in the samples;   (C) comparing the toxic variants of TDP-43, tau, abeta and/or alpha-synuclein protein levels in the sample at each time point with the toxic variant of TDP-43, tau, abeta and/or alpha-synuclein protein levels in a normal control; and   (D) determining whether the subject has a risk of TBI in accordance with the result of step (C);   
       wherein a subject having elevated toxic variants of TDP-43 tau, abeta and/or alpha-synuclein protein has a high risk of TBI. 
     
     
         12 . The method of  claim 11 , wherein a sample is obtained from the subject within 6 hours post-injury. 
     
     
         13 . The method of  claim 11 , wherein a sample is obtained from the subject about 12 to 36 hours post-injury. 
     
     
         14 . The method of  claim 11 , wherein a sample is obtained from the subject about 5 to 10 days post injury. 
     
     
         15 . The method of  claim 11 , wherein a sample is obtained from the subject about 2 to 4 weeks days post injury. 
     
     
         16 . The method of  claim 11 , wherein the samples and the normal control are blood product samples or cerebrospinal fluid (CSF) samples. 
     
     
         17 . The method of  claim 16 , wherein the blood product is serum. 
     
     
         18 . The method of  claim 11 , wherein the detecting in step (B) is by means of a ligand specific for the protein. 
     
     
         19 . The method of  claim 18 , wherein the ligand is an antibody. 
     
     
         20 . The method of  claim 18 , wherein the ligand is a designed ankyrin repeat protein (DARPin). 
     
     
         21 . The method of  claim 11 , wherein protein levels are detected by means of ELISA. 
     
     
         22 . The method of  claim 20 , wherein the DARPin is encoded by a sequence having at least 90% sequence identity of any one of SEQ ID NO: 7-28. 
     
     
         23 . The method of  claim 20 , wherein the DARPin is encoded by a sequence having 100% sequence identity of any one of SEQ ID NO: 7-28. 
     
     
         24 . A method for measuring the presence of a biomarker in a human sample from a patient having traumatic brain injury (TBI), the improvement comprising measuring the levels of toxic variants of TDP-43, tau, abeta and/or alpha-synuclein in the sample for use in predicting the amount of neuronal damage, and/or susceptibility to neurodegenerative disease in a subject.