Antibodies and Methods for the Detection of Cell Death

Abstract

Disclosed herein are antibodies having binding specificity to the amino acid sequences Ala Ser Ser Gly Leu Thr Val Glu Val Asp (SEQ ID NO:1) and Thr Val Glu Val Asp (SEQ ID NO:14), and methods of detecting cell death in a sample, comprising contacting the sample with a first antibody specific for a C-terminal amino acid sequence Ala Ser Ser Gly Leu Thr Val Glu Val Asp (SEQ ID NO:1) or Thr Val Glu Val Asp (SEQ ID NO:14) of a CK18 protein fragment having a C-terminal amino acid sequence of Val Glu Val Asp (SEQ ID NO:2) and a second antibody that specifically binds an epitope that is present in both full-length CK18 and the CK18 protein fragment, and that does not overlap with SEQ ID NO:1 or SEQ ID NO:14, under conditions such that the CK18 protein fragment present in the sample specifically binds to the first antibody and the second antibody, wherein one of the antibodies is bound to a solid support and the other antibody is bound to a detection moiety capable of producing a signal; optionally removing any unbound or excess material; and detecting the signal from the detection moiety, wherein the signal is positively correlated with the presence of the CK18 protein fragment in the sample.

What is claimed is: 
     
         1 . An antibody wherein the antibody has binding specificity to the amino acid sequence Thr Val Glu Val Asp (SEQ ID NO:14). 
     
     
         2 . The antibody of  claim 1 , coupled to a solid support or a detection moiety. 
     
     
         3 . The antibody of  claim 2 , wherein the antibody is directly bound to a solid support or a detection moiety. 
     
     
         4 . The antibody of  claim 2 , wherein the antibody is indirectly bound to a solid support or a detection moiety. 
     
     
         5 . The antibody of  claim 2 , wherein the detection moiety is selected from the group consisting of electrochemiluminescence labels or compounds, chemiluminescent compounds, enzyme labels, fluorophores, chromogenic compounds, radiolabels, catalysts, colorimetric, labeled antibodies, a latex particle, a magnetic particle, a paramagnetic particle, a radioactive element, fluorescent dyes, phosphorescent dyes, dye crystals, gold particles, silver colloidal particles, selenium colloidal particles, metal chelates, coenzymes, electro active groups, oligonucleotides and stable radicals. 
     
     
         6 . The antibody of  claim 5 , wherein the metal chelate is a ruthenium, an osmium, or a europium metal chelate. 
     
     
         7 . The antibody of  claim 2 , wherein the solid support is selected from the group consisting of a bead, a plate, a glass surface, a plastic surface, a metal surface, a polystyrene surface, a nitrocellulose surface, a microparticle and a nanoparticle surface. 
     
     
         8 . The antibody of  claim 7 , wherein the solid support is a bead. 
     
     
         9 . The antibody of  claim 1 , wherein the antibody comprises a variable region of the heavy chain amino acid sequence of SEQ ID NO:5, and a variable region of the light chain amino acid sequence of SEQ ID NO:7. 
     
     
         10 . The antibody of  claim 9 , coupled to a solid support or a detection moiety. 
     
     
         11 . The antibody of  claim 10 , wherein the antibody is directly bound to a solid support or a detection moiety. 
     
     
         12 . The antibody of  claim 10 , wherein the antibody is indirectly bound to a solid support or a detection moiety. 
     
     
         13 . The antibody of  claim 10 , wherein the detection moiety is selected from the group consisting of electrochemiluminescence labels or compounds, chemiluminescent compounds, enzyme labels, fluorophores, chromogenic compounds, radiolabels, catalysts, colorimetric, labeled antibodies a latex particle, a magnetic particle, a radioactive element, fluorescent dyes, phosphorescent dyes, dye crystallites, gold particles, silver colloidal particles, selenium colloidal particles, metal chelates, coenzymes, electro active groups, oligonucleotides and stable radicals. 
     
     
         14 . The antibody of  claim 13 , wherein the metal chelate is a ruthenium, an osmium, or a europium metal chelate. 
     
     
         15 . The antibody of  claim 10 , wherein the solid support is selected from the group consisting of a bead, a plate, a glass surface, a plastic surface, a metal surface, a polystyrene surface, a nitrocellulose surface, a microparticle and a nanoparticle surface. 
     
     
         16 . The antibody of  claim 15 , wherein the solid support is a bead.