Methionine sulfoxide antibodies

Abstract

Antibodies specific for methionine sulfoxide residues on proteins are provided. The antibodies are prepared using methionine-rich zein proteins, which are oxidized, as antigens.

1 . An isolated antibody that specifically binds a methionine sulfoxide of a polypeptide.  
   
   
       2 . The antibody of  claim 1  wherein said antibody is polyclonal.  
   
   
       3 . The antibody of  claim 1  wherein said polypeptide is selected from the group consisting of oxidized glutamine synthestase and oxidized glyceraldehyde 3-phosphate dehydrogenase.  
   
   
       4 . The antibody of  claim 1  wherein said polypeptide is selected from the group consisting of oxidized calmodulin, Ikappa B, and voltage-dependent K(+) channel.  
   
   
       5 . The antibody of  claim 1  wherein said polypeptide wherein said polypeptide is an oxidized zein protein.  
   
   
       6 . The antibody of  claim 5  wherein at least 23% of amino acid residues of said zein protein comprise methionine.  
   
   
       7 . A method of making antibodies with the specificity of the antibody of  claim 1 , the method comprising: 
 a) oxidizing a zein protein to provide a methionine sulfoxide containing zein protein;    b) immunizing an animal with the oxidized zein protein or an immunogenic fragment thereof, under conditions to elicit an antibody response; and    c) isolating antibody-producing cells from the animal.    
   
   
       8 . The method of  claim 7  further comprising fusing the antibody producing cells with immortalized cells to form monoclonal antibody-producing hybridoma cells.  
   
   
       9 . A kit comprising the isolated antibody as set forth in  claim 1  for screening for expression of a polypeptide containing methionine sulfoxide in a cell.  
   
   
       10 . A diagnostic test for a condition or disease associated with the expression of methionine sulfoxide containing polypeptides in a biological sample, the method comprising: 
 a) combining the biological sample with an antibody of  claim 1 , under conditions suitable for the antibody to bind the polypeptide and form an antibody:polypeptide complex, and    b) detecting the complex, wherein the presence of the complex correlates with the presence of the polypeptide in the biological sample.    
   
   
       11 . The test of  claim 10  wherein said antibody is polyclonal.  
   
   
       12 . The test of  claim 10  wherein said polypeptide is selected from the group consisting of oxidized glutamine synthestase and oxidized glyceraldehyde 3-phosphate dehydrogenase.  
   
   
       13 . The test of  claim 10  wherein said polypeptide is selected from the group consisting of oxidized calmodulin, Ikappa B, and voltage-dependent K(+) channel.