Biomarker detection process and assay of neurological condition

Abstract

The subject invention provides a robust, quantitative, and reproducible process and assay for diagnosis of a neurological condition in a subject. The invention provides measurement of two or more biomarkers in a biological fluid such as CSF or serum resulting in a synergistic mechanism for determining the extent of neurological damage in a subject with an abnormal neurological condition and for discerning subtypes thereof or tissue types subjected to damage.

1 . A process for determining the neurological condition of a subject or cells from the subject comprising:
 measuring a sample obtained from the subject or cells from the subject at a first time for a quantity of a first biomarker selected from the group of GFAP, UCH-L1, NSE, MAP2, S100b, or a SBDP, and a quantity of at least one additional neuroactive biomarker; and   comparing the quantity of said first biomarker and the quantity of said at least one additional neuroactive biomarker to normal levels of said first biomarker and said at least one additional neuroactive biomarker to determine the neurological condition of the subject.   
     
     
         2 . The process of  claim 1  wherein the sample is cerebrospinal fluid or blood serum. 
     
     
         3 . The process of  claim 1  wherein the sample is a culture of the cells exposed to a drug candidate or an environmental contaminant. 
     
     
         4 . The process of  claim 1  wherein said at least one additional neuroactive biomarker is GFAP, UCH-L1, NSE, SBDP150, SBDP145, SBDP120, S100b, MAP2, MAP1, MAP3, MAP4, MAPS, MBP, Tau, Neurofilament protein (NF), Cannabinoid Receptor CB, CAM, Synaptic protein, CRMP, iNOS, NeuN, CNPase, Neuroserpin, alpha-internexin, LC3, Neurofascin, EAAT, Nestin, Cortin-1, or BIII-Tubulin. 
     
     
         5 . The process of  claim 1  wherein is at least one additional neuroactive biomarker is one of GFAP, UCH-L1, NSE, SBDP150, SBDP150i, SBDP145, SBDP120, or MAP2. 
     
     
         6 . The process of  claim 1  further comprising measuring a second quantity of said first biomarker and a second quantity of said at least one additional neuroactive biomarker at a second time to yield a kinetic profile for said first biomarker and said at least one additional neuroactive biomarker. 
     
     
         7 . The process of  claim 1  further comprising comparing the quantity of said first biomarker and the quantity of said at least one additional neuroactive biomarker between normal levels in the subject to other individuals of the same gender as the subject. 
     
     
         8 . The process of  claim 1  wherein said at least one additional neuroactive biomarker is GFAP. 
     
     
         9 . The process of  claim 7  wherein said first biomarker is UCH-L1 and determined if the subject or cells from the subject has been exposed to some degree of traumatic brain injury ranging from mild to severe. 
     
     
         10 . The process of  claim 9  further comprising predicting mortality based on the quantity of UCH-L1 and the quantity of GFAP. 
     
     
         11 . The process of  claim 9  wherein mild traumatic brain injury and moderate traumatic brain injury have detection cutoffs for UCH-L1 and GFAP in serum of 0.39 ng/ml and 1.4 ng/ml, respectively. 
     
     
         12 . The process of  claim 1  wherein the at least one additional neuroactive biomarker is S100b. 
     
     
         13 . The process of  claim 1  wherein the at least one additional neuroactive biomarker is a SBDP of SBDP150, SBDP150i, SBDP145, or SBDP120. 
     
     
         14 . The process of  claim 1  wherein the at least one additional neuroactive biomarker is NSE. 
     
     
         15 . The process of  claim 1  wherein the at least one additional neuroactive biomarker is a MAP of MAP2, MAP1, MAP3, MAP4, or MAPS. 
     
     
         16 . The process of any of  claims 1  to  15  wherein the first biomarker is UCH-L1. 
     
     
         17 . An assay for determining the neurological condition of a subject or cells from the subject comprising:
 a substrate for holding a sample isolated from the subject or the cells;   a first biomarker specifically binding agent wherein a first biomarker is one of GFAP, UCH-L1, NSE, MAP2, S100b, or a SBDP;   an agent specifically binding at least one additional neuroactive biomarker; and   printed instructions for reacting said first biomarker specific agent with a first portion of the sample so as to detect an amount of said first biomarker and reacting said at least one additional neuroactive biomarker specific agent with a second portion of the sample and said at least one additional neuroactive biomarker in the sample so as to detect an amount of said at least one additional neuroactive biomarker to determine the neurological condition of the subject according to the process of  claim 1 .   
     
     
         18 . The assay of  claim 17  wherein the first biomarker specific agent is one of anti-GFAP antibody, anti-UCH-L1 antibody, anti-NSE antibody, anti-MAP2 antibody, or an anti-SBDP antibody. 
     
     
         19 . The assay of  claim 17  wherein the agent specifically binding at least one additional neuroactive biomarker binds one of GFAP, NSE, SBDP, SBDP150, SBDP150i, SBDP145, SBDP120, S100b, MAP2, MAP1, MAP3, MAP4, MAPS, MBP, Tau, Neurofilament protein (NF), Cannabinoid Receptor CB, CAM, Synaptic protein, CRMP, iNOS, NeuN, CNPase, Neuroserpin, alpha-internexin, LC3, Neurofascin, EAAT, Nestin, Cortin-1, or BIII-Tubulin. 
     
     
         20 . The assay of  claim 17  wherein the agent specifically binding at least one additional neuroactive biomarker binds GFAP. 
     
     
         21 . The assay of  claim 17  wherein the agent specifically binding at least one additional neuroactive biomarker binds S100b. 
     
     
         22 . The assay of  claim 17  wherein the agent specifically binding at least one additional neuroactive biomarker binds a SBDP of SBDP150, SBDP150i, SBDP145, or SBDP120. 
     
     
         23 . The assay of any of  claims 17  to  22  wherein the first biomarker specific agent is anti-UCH-L1 antibody. 
     
     
         24 . The assay of  claim 17  wherein said first biomarker specifically binding agent and said agent specifically binding at least one additional neuroactive biomarker are both bound to the substrate. 
     
     
         25 . The assay of  claim 24  wherein said first biomarker specifically binding agent and said agent specifically binding at least one additional neuroactive biomarker are both bound to the substrate with spatial overlap. 
     
     
         26 . The assay of  claim 17  wherein the first portion and the second portion of the sample and the second portion of the sample are the same, and detection of the amount of said first biomarker and the amount of said at least one additional neuroactive biomarker occurs simultaneously. 
     
     
         27 . The assay of  claim 26  wherein detection of the amount of said first biomarker and the amount of said at least one additional neuroactive biomarker occurs with spatial overlap. 
     
     
         28 . The assay of any of  claims 17  or  24  to  26  further comprising a separate first biomarker detection species for said first biomarker and a separate discernable at least one additional neuroactive biomarker detection species for said at least one additional neuroactive biomarker. 
     
     
         29 . The assay of  claim 17  wherein the neurological condition is stroke, ischemic stroke, hemorrhagic stroke, or subarachnoid hemorrhage (SAH). 
     
     
         30 . The assay of  claim 17  wherein the neurological condition is mild traumatic brain injury or moderate traumatic brain injury. 
     
     
         31 . A process for determining if a subject has suffered mild traumatic brain injury or moderate traumatic brain injury in an event comprising:
 measuring a sample obtained from the subject or cells from the subject at a first time after the event for a quantity of GFAP; and   comparing the quantity of said GFAP to normal levels of GFAP in a control to determine if the subject has suffered mild traumatic brain injury or moderate traumatic brain injury in the event.   
     
     
         32 . The process of  claim 31  wherein the first time is within 48 hours of the event. 
     
     
         33 . The process of  claim 31  wherein the sample is blood serum. 
     
     
         34 . The process of any of  claims 31  to  33  wherein a mean GFAP value for the subject having suffered suffered mild traumatic brain injury or moderate traumatic brain injury in the event is approximately 0.28 nanograms per milliliter of the sample.