Detection of specific nitrated markers

Abstract

Methods are described for improving the diagnostic possibilities of diseases where oxidative NO-modifications occur, for example inflammatory conditions, cancer, Parkinson's or Alzheimer's disease, and to provide means of monitoring the effects of therapeutical measures taken towards such diseases. The invention enables the detection of disease specific catabolic markers related to oxidative NO-modifications, utilizing an immunoassay comprising antibodies directed against nitrated and non-nitrated epitopes characteristic of a specific protein.

1 - 26 . (canceled) 
     
     
         27 . A kit for performing an immunoassay comprising the steps of: detecting binding of an immunological binding partner which is immunoreactive with one or more aromatic amino acid residues in nitrated form of an amino acid sequence which is characteristic of a specific protein; the kid comprising: an immunological binding partner which is specifically reactive with the nitrated form of an aromatic amino acid residue in the context of an amino acid sequence which is characteristic of a specific protein; and means for detecting binding of said binding partner and said protein or a fragment thereof. 
     
     
         28 . The kit as claimed in  claim 27 , wherein the immunological binding partner is an immunological binding partner specifically reactive with the nitrated form of an aromatic amino acid residue in the context of an amino acid sequence which is characteristic of a specific protein. 
     
     
         29 . The kit as claimed in  claim 27 , including an immunological binding partner which is reactive with the said amino acid sequence in non-nitrated form. 
     
     
         30 . The kit as claimed in  claim 27 , comprising a peptide reactive with a said immuno-logical binding partner. 
     
     
         31 . An immunological binding partner specifically reactive with the nitrated form of an aromatic amino acid residue in the context of an amino acid sequence which is characteristic of a specific protein. 
     
     
         32 . The immunological binding partner as claimed in  claim 31 , having binding specificity for an epitope contained in the amino acid sequence HRGY:NO 2 PGLDG (SEQ ID NO: 6) which epitope contains the amino acid residue Y:NO 2  and is characteristic of collagen type II. 
     
     
         33 . The immunological binding partner as claimed in  claim 32 , which is an antibody raised against a peptide having the sequence (X aa ) m HRGY:NO 2 PGLDG(X aa ) n  (SEQ ID NO: 4), where-in X denotes any amino acid or derivative thereof and m and n are independent integers of from 1 to 10, or is a fragment of such an antibody. 
     
     
         34 . The immunological binding partner as claimed in  claim 31 , having binding specificity for an epitope contained in the amino acid sequence LQY:NO 2 MRA (SEQ ID NO: 7) which epitope contains the amino acid residue Y:NO 2  and is characteristic of collagen type II. 
     
     
         35 . The immunological binding partner as claimed in  claim 34 , which is an antibody raised against a peptide having the sequence (X aa ) m LQY:NO 2 MRA(X aa ) n  (SEQ ID NO: 5), wherein X denotes any amino acid or derivative thereof and m and n are independent integers of from 1 to 10, or is a fragment of such an antibody. 
     
     
         36 . The immunological binding partner as claimed in  claim 31 , which is a monoclonal antibody or fragment thereof. 
     
     
         37 . A cell line producing a monoclonal antibody or fragment thereof as claimed in  claim 36 .