Methods of determining a treatment protocol for and/or a prognosis of a patient's recovery from a brain injury

Abstract

The present invention, in some embodiments, generally relates to methods of determining a treatment protocol for and/or a prognosis of a patient's recovery from a brain injury. In some embodiments, the brain injury results from a hypoxic event. In some embodiments, methods are provided for determining a measure of the concentration of tau protein in a patient sample containing or suspected of containing tau protein.

What is claimed: 
     
         1 - 91 . (canceled) 
     
     
         92 . A method for determining a measure of the concentration of a biomarker generated in the brain in a volume of bodily fluid obtained from a subject, comprising performing an assay on a sample containing or derived from the volume of bodily fluid to determine a measure of the concentration of the biomarker in the bodily fluid, wherein:
 the volume of bodily fluid contains or is suspected of containing the biomarker,   the volume of bodily fluid comprises blood or a blood component selected from plasma and serum, and   the limit of quantification of the biomarker of the assay is less than about 0.2 pg/mL.   
     
     
         93 . The method of  claim 92 , wherein the biomarker comprises a protein. 
     
     
         94 . The method of  claim 92 , wherein the biomarker comprises neuron specific neuronal enolase (NSE), β-site aPP-cleaving enzyme 1 (BACe1), S100B, myelin basic protein (MBP), growth associated protein 43, glutamine synthetase, glial fibrillary acid protein (GFAP), a glycine transporter, a neuron specific glycoprotein, calpain, neurofibrillary protein, heat shock protein 72, beta-amyloid precursor proteins, calbindin D-28K, proteolipid protein, myeline associated glycoprotein, neurofilament H, a creatine kinase protein, a tau protein, and/or an endothelium membrane protein. 
     
     
         95 . The method of  claim 92 , wherein the assay comprises:
 i. suspending a plurality of capture objects that each include a binding surface having affinity for the biomarker in the sample,   ii. immobilizing at least some of the biomarker molecules with respect to at least some of the plurality of capture objects suspended in the sample such that at least some of the capture objects associate with at least one molecule of the biomarker from the volume of bodily fluid,   iii. interrogating at least some of the capture objects associated with at least one molecule of the biomarker from the volume of bodily of fluid; and   iv. determining a measure of the biomarker concentration in the volume of bodily fluid based at least in part on the interrogating step performed in step iii.   
     
     
         96 . The method of  claim 95 , wherein the plurality of capture objects comprises a plurality of beads. 
     
     
         97 . The method of  claim 95 , wherein the interrogating in step iii comprises exposing the at least some of the capture objects associated with at least one molecule of the biomarker to electromagnetic radiation and detecting emission. 
     
     
         98 . The method of  claim 92 , wherein the assay comprises:
 i. exposing a plurality of capture objects that each include a binding surface having affinity for the biomarker to the sample;   ii. immobilizing at least some of the biomarker molecules with respect to the plurality of capture objects such that at least some of the capture objects associate with at least one molecule of the biomarker and a statistically significant fraction of the capture objects do not associate with any molecules of the biomarker;   iii. determining a measure indicative of the number or fraction of the plurality of capture objects that associated with a molecule of the biomarker in step ii; and   iv. determining a measure of the biomarker concentration in the volume of bodily fluid based at least in part on the measure indicative of the number or fraction determined in step iii.   
     
     
         99 . The method of  claim 98 , wherein the plurality of capture objects comprises a plurality of beads. 
     
     
         100 . The method of  claim 92 , wherein the assay comprises spatially segregating molecules of the biomarker into a plurality of locations, wherein a statistically significant fraction of the locations contain a biomarker molecule and a statistically significant fraction of the locations do not contain any biomarker molecules. 
     
     
         101 . The method of  claim 100 , wherein the plurality of locations comprises a plurality of reaction vessels. 
     
     
         102 . The method of  claim 101 , wherein the reaction vessels have a volume between about 10 attoliters and about 100 picoliters. 
     
     
         103 . The method of  claim 102 , wherein the reaction vessels have a volume between about 1 femtoliter and about 1 picoliter. 
     
     
         104 . The method of  claim 92 , wherein the assay comprises:
 providing a plurality of capture objects, at least some of which are associated with at least one molecule of the biomarker from the sample;   passing at least some of the capture objects associated with at least one molecule of the biomarker from the sample by a detection system;   individually interrogating, during the passing step, the at least some of the capture objects passed by the detection system; and   determining a measure of the biomarker concentration in the volume of bodily fluid based at least in part on the interrogating step.   
     
     
         105 . The method of  claim 104 , wherein at least some of the plurality of capture objects provided are associated with at least one molecule of the biomarker and a statistically significant fraction of the plurality of capture objects provided are not associated with any molecules of the biomarker. 
     
     
         106 . The method of  claim 105 , wherein the step of determining a measure of the biomarker concentration comprises determining a measure indicative of a number or fraction of interrogated capture objects determined to be associated with at least one molecule of the biomarker. 
     
     
         107 . The method of  claim 106 , wherein at least some of the capture objects passed by the detection system are in single file. 
     
     
         108 . The method of  claim 107 , wherein the detection system is an optical detection system. 
     
     
         109 . The method of  claim 108 , wherein the capture objects comprise or are each contained within a liquid droplet that is immiscible within a fluid in which the droplet is suspended during at least the interrogating step. 
     
     
         110 . The method of  claim 92 , wherein the bodily fluid is blood. 
     
     
         111 . The method of  claim 92 , wherein the bodily fluid is whole blood. 
     
     
         112 . The method of  claim 92 , wherein the bodily fluid is plasma. 
     
     
         113 . The method of  claim 92 , wherein the bodily fluid is serum. 
     
     
         114 . The method of  claim 92 , wherein the limit of quantification of the biomarker is less than about 0.02 pg/mL.