Detection of Degradation Products of Feline NT-proBNP

Abstract

A method for determining the amount of NT-proBNP in blood samples from felines. The method includes detecting degradation products of feline NT-proBNP by various methods, including using antibodies, kits and devices.

What is claimed is: 
     
         1 . A method for determining the amount of NT-proBNP in a sample from a feline comprising:
 a. obtaining a blood, serum or plasma sample from a feline subject;   b. analyzing the sample by contacting the sample with a first antibody that binds a polypeptide selected from the group consisting of ASEASAIQELLDGLRDTVSEL (SED ID NO: 5), LGPLQQGHSPAESWEAQEEPPARVLAPHDNVLR (SEQ ID NO:9) and LQQGHSPAESWEAQEEPPARVLAPHDNVLR (SEQ ID NO:10);   c. detecting the amount of binding between the antibody and the polypeptide and the polypeptide, wherein the binding is detectable after at least 96 hours at room temperature   d. associating the amount of binding to the amount of NT-proBNP in the sample.   
     
     
         2 . The method of  claim 2 , wherein the determining further comprises: contacting a complex formed by the first antibody and the NT-proBNP in the sample with a second antibody specific for feline NT-proBNP, wherein the second antibody is directly or indirectly conjugated to a label; and detecting the amount of the label associated with the complex, wherein the first antibody and the second antibody do not compete for binding to the same epitope on NT-proBNP. 
     
     
         3 . The method of  claim 2 , wherein the second antibody is specific for one of SEQ ID NO:5, SEQ ID NO:9 and SEQ ID NO: 10. 
     
     
         4 . The method of  claim 1 , further comprising contacting a standard comprising recombinant feline NT-proBNP with the first antibody, detecting the binding of the standard to the first antibody, and comparing the amount of binding of the first antibody to the NT-proBNP in the sample to the amount of binding of the first antibody to the standard. 
     
     
         5 . The method of  claim 2 , further comprising contacting a standard comprising recombinant feline NT-proBNP with the first antibody and the second antibody, and comparing a signal from the label of the second antibody that is bound to the NT-proBNP in the sample to the signal from the label of the second antibody that is bound to the standard. 
     
     
         6 . The method of  claim 1 , wherein the first antibody is a monoclonal antibody raised against a polypeptide selected from a sequence consisting essentially of one of SEQ ID NO:5, SEQ ID NO:9 and SEQ ID NO: 10. 
     
     
         7 . The method of  claim 3 , wherein the second antibody is monoclonal antibody. 
     
     
         8 . The method of  claim 1 , further comprising analyzing the sample after at least 192 hours at room temperature. 
     
     
         9 . The method of  claim 1 , wherein the first antibody is specific for SEQ ID NO: 5. 
     
     
         10 . The method of  claim 1 , wherein the first antibody is specific for SEQ ID NO:9. 
     
     
         11 . The method of  claim 1 , wherein the first antibody is specific for SEQ ID NO:10. 
     
     
         12 . A method for determining the presence or amount of feline NT-proBNP in a sample comprising:
 a. obtaining a blood, serum or plasma sample from a feline;   b. forming a complex comprising NT-proBNP in the sample, a first antibody that specifically a polypeptide selected from the group consisting of SEQ ID NO:5, SEQ ID NO:9 and SEQ ID NO: 10, and a second antibody that binds to feline NT-proBNP, wherein one of the first antibody and second antibody is bound to solid phase and the other of the first antibody and second antibody is bound to a label, wherein the complex can be formed after more than about 96 hours after obtaining the sample at room temperature;   c. detecting the presence or amount of the label on the solid phase, thereby determining the presence or amount of feline NT-proBNP in the sample.   
     
     
         13 . The method of  claim 12 , wherein the first antibody is a monoclonal antibody. 
     
     
         14 . The method of  claim 12 , wherein the second antibody is a monoclonal antibody. 
     
     
         15 . The method of  claim 12 , wherein the first antibody is specific for SEQ ID NO: 5. 
     
     
         16 . The method of  claim 12 , wherein the first antibody is specific for SEQ ID NO:9. 
     
     
         17 . The method of  claim 12 , wherein the first antibody is specific for SEQ ID NO:10. 
     
     
         18 . The method of  claim 12 , further comprising forming the complex after at least 192 hours at room temperature.