Methods of determining stabilization compounds for predictive biomarkers

Abstract

Methods of determining stabilization compounds for predictive biomarkers. According to at least one embodiment of a method of the present disclosure, the method comprises introducing a diagnostic marker and a stabilization agent to a detection platform, mixing the diagnostic marker and stabilization agent, and determining their binding characteristics. Additionally, the method may further comprise generating a report based on the binding characteristics and delivering the report to a recipient.

1 . A method for analyzing stabilization agents, the method comprising the steps of:
 introducing a diagnostic marker into each of a plurality of detection sites of a detection platform, the detection platform comprising:
 the plurality of detection sites each capable of receiving a diagnostic marker, a stabilization agent, and a detection agent; 
 a detection device capable of determining a binding characteristic between the detection agent and the diagnostic marker in the detection sites; 
 a computer database capable of receiving a plurality of binding characteristics, the plurality of binding characteristics comprising at least one binding property of a diagnostic marker to a detection agent; 
 a processor operably coupled to the computer database and the detection device, the processor having and executing a software program operational to:
 determine the binding characteristic of the detection agent to the diagnostic marker in each of the plurality of detection sites; 
 compare the binding characteristic among each of the plurality of detection sites to order the level of effect of the stabilizing agents assayed on the binding characteristic between the detection agent and the diagnostic agent; 
 generate a binding record using the compared binding characteristics; and 
 deliver the binding record to a recipient; 
 
   introducing the stabilization agent to each of the plurality of detection sites containing the diagnostic marker;   combining the detection agent with the stabilization agent and diagnostic marker in the detection sites;   comparing the binding characteristic with the processor among each of the plurality of detection sites to order the level of effect of the stabilizing agents assayed on the binding characteristic between the detection agent and the diagnostic agent;   generating a binding report using the compared binding characteristics; and   delivering the binding report to a recipient.   
     
     
         2 . The method of  claim 1 , wherein the stabilizing agent is selected from the group consisting of a protease inhibitor, a DNase inhibitor, and a RNase inhibitor. 
     
     
         3 . The method of  claim 1 , wherein the stabilizing agent prevents the degradation or inactivation of the diagnostic marker for at least a twenty four hours period following interaction with the diagnostic agent. 
     
     
         4 . The method of  claim 1 , wherein the diagnostic marker is selected from the group consisting of a protein, a glycoprotein, a nucleic acid, an enzyme, an enzyme inhibitor, and a metabolite. 
     
     
         5 . The method of  claim 1 , wherein the stabilizing agent is useful to completely or substantially inactivate an enzyme selected from the group consisting of an amylase, a lysozyme, a peroxidase, a glycosidase, an esterase, a protease, and a peptidase. 
     
     
         6 . The method of  claim 1 , wherein the stabilizing agent is selected from the group consisting of Fixanal® Buffer 6.0 (Sigma-Aldrich Co.), acetic acid, aluminum hydroxide bentonite, aluminum sulfate hydrate, aluminum potassium sulfate dodecahydrate, benzoic acid, caffeine, and 3-tert-butyl-hydroxyanisole, or a combination thereof. 
     
     
         7 . The method of  claim 1 , wherein the stabilizing agent comprises a plurality of stabilizing agents each present in approximately the same concentration. 
     
     
         8 . The method of  claim 1 , wherein the stabilizing agent is capable of inhibiting degradation of the diagnostic marker to an inhibitory degree, wherein the inhibitory degree is selected from the group consisting of at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, and at least about 99%. 
     
     
         9 . The method of  claim 1 , wherein the stabilizing agent has a concentration selected from the group consisting of about 200 parts per million (ppm) to about 2000 ppm, about 400 ppm to about 1600 ppm, about 600 ppm to about 1400 ppm, about 800 ppm to about 1200 ppm, and about 400 ppm to about 600 ppm. 
     
     
         10 . The method of  claim 1 , wherein the stabilizing agent is able to inhibit the degradation or inactivation of the diagnostic marker for an inhibitory period selected from the group consisting of at least one minute, at least about five minutes, at least about ten minutes, at least about fifteen minutes, at least about thirty minutes, at least about one hour, at least about two hours, at least about four hours, and at least about eight hours. 
     
     
         11 . The method of  claim 1 , wherein the diagnostic marker is selected from the group consisting of Aldose reductase, Angiogenin, Annexin A1, B-cell activating factor (BAFF), B-cell lymphoma 2 (BCL2)-like 2, Beta Human chorionic gonadotropin, Ca15-3, Calcyclin, Calvasculin, Cancer Antigen CA 19-9, Cancer Antigen CA 15-3, Cathepsin D, Caveolin-1, Chromogranin A, Alpha-crystallin B chain (CRYAB), Endostatin, Eotaxin-2, Epithelial cell adhesion molecule (EpCAM), Ezrin, fatty acid binding protein 4 (FABP4), Galectin-3, γ-glutamylcysteine ligase regulatory chain (GCLR), Gelsolin, Glucose 6-phosphate (G6P), Glycoprotein 130 (gp130), Glutathione S-transferase Mu 1 (GSTM1), Hepsin, High-mobility group protein B1 (HMGB-1), Insulin-like growth factor binding protein 1 (IGFBP-1), Insulin-like growth factor binding protein 4 (IGFBP-4), Insulin-like growth factor binding protein 5 (IGFBP-5), Insulin-like growth factor binding protein 6 (IGFBP-6), LGL, latency associated peptide (LAP), macrophage stimulating protein (MSP), MHC class I polypeptide-related sequence A (MICA), Nucleoside diphosphate kinase B (NME2), Neuron-specific Enolase (NSE), Osteopontin, Osteoprotegerin, Pepsinogen, Peroxiredoxin, Phosphoserine aminotransferase (PSAT1), Prostate Specific Antigen, Receptor tyrosine-protein kinase erbB-3 (ErbB3), Serpin B3, Vascular smooth muscle cell growth factor R2 (VSGF R2/KDR), Vascular endothelial growth factor R3 (VEGF R3/Flt-4), Thyroglobulin, Tyrosine kinase with immunoglobulin-like and EGF-like domains 2 (TIE-2), Tissue plasminogen activator (tPA), Transforming growth factor beta (TGF-β1), Tumor necrosis factor receptor 1 (TNF-R1), urokinase-type Plasminogen Activator (uPA), urokinase-type Plasminogen Activator Receptor (uPAR), BrcaI, BrcaII, kallikreins, e-cadherin, Hox peptide, and Engrailed-2.