Methods and products for in vivo enzyme profiling

Abstract

The present invention relates to methods and products associated with in vivo enzyme profiling. In particular, the invention relates to methods of in vivo processing of exogenous molecules followed by detection of signature molecules as representative of the presence of active enzymes associated with diseases or conditions. The invention also relates to products, kits, and databases for use in the methods of the invention.

1 - 88 . (canceled) 
     
     
         89 . A method comprising:
 introducing a diagnostic reagent, wherein said diagnostic reagent comprises a carrier linked to a signature molecule by a peptide, wherein said peptide is cleaved by a protease wherein said cleavage releases said signature molecule, and wherein said diagnostic reagent is not coupled to a chip;   performing an assay to detect said signature molecule cleaved from said carrier; and   evaluating a disease based on a quantifiable amount of said signature molecule,   wherein evaluating said disease is based on an amount of said released signature molecule, and   wherein said disease is a blood pathology.   
     
     
         90 . The method of  claim 89 , wherein said detection is conducted in a solution. 
     
     
         91 . The method of  claim 89 , wherein said signature molecule comprises a peptide sequence, a nucleic acid, a small molecule, a fluorophore, a carbohydrate, a particle, a radiolabel, a MRI-active compound, an inorganic material, and/or an organic material. 
     
     
         92 . The method of  claim 91 , wherein said signature molecule comprises encoded characteristics to facilitate optimal detection. 
     
     
         93 . The method of  claim 92 , wherein said encoded characteristics are specific to detection using mass spectrometry. 
     
     
         94 . The method of  claim 91 , wherein said diagnostic reagent further comprises a quencher configured to quench the fluorophore. 
     
     
         95 . The method of  claim 89 , wherein said cleavage of said peptide generates a detectable signal. 
     
     
         96 . The method of  claim 89 , wherein said protease cleaves said peptide in vivo. 
     
     
         97 . The method of  claim 89 , wherein said protease cleaves said peptide in vitro. 
     
     
         98 . The method of  claim 89 , wherein said protease cleaves said peptide ex vivo. 
     
     
         99 . The method of  claim 89 , wherein evaluating said disease comprises analyzing a disease metric of said disease. 
     
     
         100 . The method of  claim 99 , wherein said disease metric includes a genetic predisposition for development of a particular disease. 
     
     
         101 . The method of  claim 99 , wherein said evaluating of said disease is non-invasive to a subject. 
     
     
         102 . The method of  claim 89 , wherein said protease comprises MMP-2, MMP-7, MMP-8, MMP-9, MMP-14, thrombin, factor Xa, tissue factor, cathepsin B, or a combination thereof. 
     
     
         103 . The method of  claim 89 , wherein said signature molecule is detected at a level above a normal value. 
     
     
         104 . The method of  claim 89 , wherein said carrier comprises a particle, and wherein said particle is a microparticle or a nanoparticle. 
     
     
         105 . A method comprising:
 introducing a diagnostic reagent, wherein said diagnostic reagent comprises a carrier linked to a signature molecule by a peptide, wherein said peptide is cleaved by a protease, wherein said cleavage releases said signature molecule, and wherein said carrier comprises a targeting agent;   performing an assay to detect said signature molecule cleaved from said carrier; and   evaluating a disease based on a quantifiable amount of said signature molecule.   
     
     
         106 . The method of  claim 105 , wherein said targeting agent is selected from the group consisting of a glycoprotein, an antibody, and a binding protein. 
     
     
         107 . The method of  claim 105 , wherein said signature molecule comprises a peptide sequence, a nucleic acid, a small molecule, a fluorophore, a carbohydrate, a particle, a radiolabel, a MRI-active compound, an inorganic material, and/or an organic material. 
     
     
         108 . The method of  claim 105 , wherein said protease comprises MMP-2, MMP-7, MMP-8, MMP-9, MMP-14, thrombin, factor Xa, tissue factor, cathepsin B, or a combination thereof.