Proteolytic digestion of cardiac troponin i

Abstract

A fluid sample container comprising a protease, when a sample fluid is placed in the fluid sample container the protease breaking a target analyte in the sample fluid into at least two peptides, the at least two peptides being smaller than the original target analyte.

1 .- 4 . (canceled) 
     
     
         5 . A device comprising:
 a sample fluid filter; and   a protease, wherein when a heparinized whole blood sample is mixed with the protease, the protease breaking a cardiac troponin in the heparinized whole blood sample into at least two peptides, the at least two peptides being smaller than the original target analyte;   wherein at least one of the at least two peptides is small enough to pass through the sample fluid filtration device; and   wherein the heparin prevents red blood cells (RBCs) in the whole blood sample from clotting, the heparin binding to individual RBCs and to at least one of the at least two peptides but less than all of the at least two peptides to form a bound compound; and   wherein the sample fluid filter prevents the bound compound from flowing there through.   
     
     
         6 .- 7 . (canceled) 
     
     
         8 . The device of  claim 5 , wherein the protease is disposed within the sample fluid filter. 
     
     
         9 . The device of  claim 5 , wherein the protease is mixed with the sample fluid in a sample container external to the sample fluid filter. 
     
     
         10 . The device of  claim 5 , wherein the protease is chymotrypsin. 
     
     
         11 . The device of  claim 10 , wherein the cTn molecule is at least one of cardiac troponin I (cTnI) or cardiac troponin T (cTnT). 
     
     
         12 .- 15 . (canceled) 
     
     
         16 . The device of  claim 5 , wherein the cardiac troponin is cardiac I (cTnI) and is broken down into at least three peptides. 
     
     
         17 . The device of  claim 16 , wherein at least one of the middle peptide fragments of cTnI has the amino acid sequence represented by SEQ ID NO:3. 
     
     
         18 . The device of  claim 16 , wherein the heparin present in the whole blood sample fluid binds to a C-terminal peptide fragment of cTnI and wherein the C-terminal peptide fragment of cTnI has the amino acid sequence represented by SEQ ID NO:4. 
     
     
         19 . The device of  claim 17 , wherein the middle peptide fragment is detected using a detection technology selected from fluorescence, chemiluminescence, electrochemical luminescence, electrochemically, and luminescent oxygen channeling immunoassay (LOCI). 
     
     
         20 . The device of  claim 16 , wherein the at least three peptides comprises at least an N-terminal peptide fragment, a C-terminal peptide fragment, and a middle peptide fragment, wherein the heparin present in the whole blood sample fluid binds to the C-terminal peptide fragment of cTnI; and
 wherein the N-terminal peptide fragment of cTnI has the amino acid sequence represented by SEQ ID NO:2.