Neural proteins as biomarkers for nervous system injury and other neural disorders

Abstract

The present invention identifies biomarkers that are diagnostic of nerve cell injury and/or neuronal disorders. Detection of different biomarkers of the invention are also diagnostic of the degree of severity of nerve injury, the cell(s) involved in the injury, and the subcellular localization of the injury.

1 . A kit comprising:
 (a) a substrate for holding a sample isolated from a subject;   (b) an agent that specifically interacts with ubiquitin C-terminal hydrolase L1 (UCH-L1);   (c) an optional additional agent that specifically interacts with at least one additional protein biomarkers upon contact with said sample; and   (d) printed instructions for reacting the agent and the optional additional agent with the sample or a portion of the sample for diagnosing a neural injury or neuronal disorder in the subject.   
     
     
         2 . The kit of  claim 1 , wherein said optional additional agent is present, and said optional additional agent interacts with said one additional protein biomarker upon contact with said sample, wherein said additional protein biomarkers is selected from the group consisting of: vesicular membrane protein p-24, synuclein, microtubule-associated protein, synaptophysin, Vimentin, Synaptotagmin, Synaptojanin-2, Synapsin2, CRMP1, 2, Amphiphysin-1, PSD95, PSD-93, Calmodulin dependent protein kinase II (CAMPK)-alpha, beta, gamma, Myelin basic protein (MBP), Myelin proteolipid protein (PLP), Myelin Oligodendrocyte specific protein (MOSP), Myelin Oligodendrocyte glycoprotein (MOG), myelin associated protein (MAG), NF-H, NF-L, NF-M, BIII-tubulin-1 and combinations thereof. 
     
     
         3 . The kit of  claim 1 , wherein said additional protein biomarkers is selected from the group consisting of: vesicular membrane protein p-24, synuclein, synaptophysin and combinations thereof. 
     
     
         4 . The kit of  claim 1 , further comprising reagents for the reacting the agent with the sample as an immunoassay. 
     
     
         5 . The kit of  claim 4 , wherein the immunoassay is an ELISA. 
     
     
         6 . The kit of  claim 1 , wherein the agent is an antibody that binds to UCH-L1. 
     
     
         7 . The kit of  claim 1 , wherein the optional agent is an antibody that binds with said at least one additional protein biomarker, the said at least one additional protein biomarker selected from the group consisting of: vesicular membrane protein p-24, synuclein, microtubule-associated protein, synaptophysin, Vimentin, Synaptotagmin, Synaptojanin-2, Synapsin2, CRMP1, 2, Amphiphysin-1, PSD95, PSD-93, Calmodulin dependent protein kinase II (CAMPK)-alpha, beta, gamma, Myelin basic protein (MBP), Myelin proteolipid protein (PLP), Myelin Oligodendrocyte specific protein (MOSP), Myelin Oligodendrocyte glycoprotein (MOG), myelin associated protein (MAG), NF—H, NF-L, NF-M, BIII-tubulin-1 and combinations thereof. 
     
     
         8 . The kit of  claim 7 , wherein said additional protein biomarkers is selected from the group consisting of: vesicular membrane protein p-24, synuclein, synaptophysin and combinations thereof. 
     
     
         9 . The kit of  claim 7 , further comprising a second antibody that binds with a second protein biomarker of said at least one additional protein biomarker. 
     
     
         10 . The kit of  claim 7 , further comprising a third antibody that binds with a third protein biomarker of said at least one additional protein biomarker. 
     
     
         11 . The kit of  claim 1  wherein said substrate is a biochip array. 
     
     
         12 . The kit of  claim 11  wherein the biochip array is a protein chip array. 
     
     
         13 . The kit of  claim 11  wherein the biochip array is a nucleic acid array. 
     
     
         14 . The kit of  claim 11  wherein the biochip array has a surface comprising a substance selected from the group consisting of: an antibody, nucleic acid, protein, peptides, amino acid probes, and a phage display library. 
     
     
         15 . A method of detecting a neural injury or neuronal disorder in a subject comprising:
 collecting a sample of a bodily fluid or a tissue in contact with neural tissue from the subject; and   analyzing said sample or a fraction thereof for an amount of ubiquitin C-terminal hydrolase L1 (UCH-L1) associated with the neural injury or neuronal disorder;   optionally analyzing a sample of the fluid or the tissue for at least one additional protein biomarker associated with the neural injury and/or neuronal disorder.   
     
     
         16 . The method of  claim 15 , wherein said at least one additional protein biomarker is selected from the group consisting of: vesicular membrane protein p-24, synuclein, microtubule-associated protein, synaptophysin, Vimentin, Synaptotagmin, Synaptojanin-2, Synapsin2, CRMP1, 2, Amphiphysin-1, PSD95, PSD-93, Calmodulin dependent protein kinase II (CAMPK)-alpha, beta, gamma, Myelin basic protein (MBP), Myelin proteolipid protein (PLP), Myelin Oligodendrocyte specific protein (MOSP), Myelin Oligodendrocyte glycoprotein (MOG), myelin associated protein (MAG), NF—H, NF-L, NF-M, BIII-tubulin-1 and combinations thereof. 
     
     
         17 . The method of  claim 15 , wherein said at least one additional protein biomarkers is selected from the group consisting of: vesicular membrane protein p-24, synuclein, synaptophysin and combinations thereof. 
     
     
         18 . The method of  claim 15 , wherein the amount decreases with recovery of the subject from the neural injury and/or neuronal disorder. 
     
     
         19 . The method of  claim 15 , wherein the amount of UCH-L1 is related to severity of the neural injury or neuronal disorder. 
     
     
         20 . The method of  claim 15 , further comprising analyzing a second sample of the bodily fluid or tissue in contact with neural tissue from the subject for a second sample amount of UCH-L1. 
     
     
         21 . The method of  claim 20  wherein said second sample amount is less than the sample amount. 
     
     
         22 . The method of  claim 20  wherein said second sample amount is more than the sample amount. 
     
     
         23 . The method of  claim 15 , wherein the neural injury or neuronal disorder is subcellular neural cell injury. 
     
     
         24 . The method of  claim 15 , wherein the neural injury or neuronal disorder is traumatic brain injury (TBI). 
     
     
         25 . The method of  claim 15  wherein said bodily fluid in contact with neural tissue is selected from the group consisting of blood, blood plasma, serum and urine. 
     
     
         26 . The method of  claim 15  wherein said UCH-L1 or one of said at least one additional protein biomarkers are detected using an immunoassay. 
     
     
         27 . The method of  claim 26  wherein the immunoassay is an ELISA. 
     
     
         28 . The method of  claim 15  wherein said at least one additional protein biomarker is two or three or four or five protein biomarkers. 
     
     
         29 . The method of  claim 15  further comprising immobilizing said at least one protein biomarker on a biochip array and laser ionizing to detect a biomarker molecular weight. 
     
     
         30 . The method of  claim 29  further comprising comparing the molecular weight against a threshold intensity that is normalized against total ion current. 
     
     
         31 . The method of  claim 29  wherein the biochip array surface comprises a substance selected from the group consisting of: an antibody, nucleic acid, protein, peptides, amino acid probes, and a phage display library.