Method of detecting cleaved snap25 in tissue samples

Abstract

Methods and compositions for detecting BoNT/A enzymatic activity in tissues or a tissue sample are described herein. The invention encompasses antibodies that bind preferentially to BoNT/A cleaved SNAP25 and is able to preferentially detect BoNT/A cleaved SNAP25, as compared to intact (non-cleaved) SNAP25, in a tissue sample.

We claim: 
     
         1 . A method of diagnosing if a tissue has been exposed to BoNT/A enzymatic activity comprising contacting a tissue sample suspected of having been exposed to BoNT/A enzymatic activity with an anti-SNAP25 antibody wherein the antibody binds preferentially to BoNT/A cleaved SNAP25; and detecting whether the anti-SNAP25 antibody bound to the tissue sample, wherein the presence of the anti-SNAP25 antibody binding to the tissue sample indicates that the tissue sample has been exposed to BoNT/A activity. 
     
     
         2 . The method of  claim 1 , wherein the antibody does not bind to full length SNAP25. 
     
     
         3 . The method of  claim 1 , wherein the antibody is able to preferentially bind to BoNT/A cleaved SNAP25 in a tissue. 
     
     
         4 . The method of  claim 1 , wherein the tissue is a biopsy sample. 
     
     
         5 . The method of  claim 1 , wherein the antibody comprises a light chain sequence of SEQID NO:1 or SEQ ID NO:2 and is a recombinant murine antibody. 
     
     
         6 . The method of  claim 1 , wherein the antibody comprises a heavy chain sequence of SEQID NO:3 or SEQID NO:4 and is a recombinant murine antibody. 
     
     
         7 . The method of  claim 1 , wherein the antibody comprises a light chain sequence of SEQID NO: 5 or SEQID NO: 6 and is a recombinant human antibody. 
     
     
         8 . The method of  claim 1 , wherein the antibody comprises a heavy chain sequence of SEQID NO:7 or SEQID NO:8 and is a recombinant human antibody. 
     
     
         9 . The method of  claim 1 , wherein the antibody binds to the same epitope of an antibody with a heavy chain and/or light chain comprising one or more sequences of SEQID NOs:1-8. 
     
     
         10 . A kit for diagnosing if a tissue has been exposed to BoNT/A enzymatic activity comprising an anti-SNAP25 antibody, wherein the antibody binds preferentially to BoNT/A cleaved SNAP25. 
     
     
         11 . The kit of  claim 10 , wherein the antibody does not bind to full length SNAP25. 
     
     
         12 . The kit of  claim 10 , wherein the antibody is able to preferentially bind to BoNT/A cleaved SNAP25 in a tissue. 
     
     
         13 . The kit of  claim 10 , wherein the tissue is a biopsy sample. 
     
     
         14 . The kit of  claim 10 , wherein the antibody comprises a light chain sequence of SEQID NO:1 or SEQ ID NO:2 and is a recombinant murine antibody. 
     
     
         15 . The kit of  claim 10 , wherein the antibody comprises a heavy chain sequence of SEQID NO:3 or SEQID NO:4 and is a recombinant murine antibody. 
     
     
         16 . The kit of  claim 10 , wherein the antibody comprises a light chain sequence of SEQID NO: 5 or SEQID NO: 6 and is a recombinant human antibody. 
     
     
         17 . The kit of  claim 10 , wherein the antibody comprises a heavy chain sequence of SEQID NO:7 or SEQID NO:8 and is a recombinant human antibody. 
     
     
         18 . The kit of  claim 10 , wherein the antibody binds to the same epitope of an antibody with a heavy chain and/or light chain comprising one or more sequences of SEQID NOs:1-8.