Anti-sulfotyrosine antibodies
View Patent ↗The invention provides anti-sulfotyrosine specific antibodies capable of detecting and isolating polypeptides that are tyrosine-sulfated. The sulfotyrosine antibodies and antibody fragments of the invention may be used to discriminate between the non-sulfated and sulfated forms of such proteins, using any number of immunological assays, such ELISAs, immunoblots, Western Blots, immunoprecipitations, and the like. Using a phage-display system, single chain antibodies (scFvs) were generated and screened against tyrosine-sulfated synthetic peptide antigens, resulting in the isolation of scFvs that specifically recognize sulfotyrosine-containing peptides and/or demonstrate sulfotyrosine-specific binding in tyrosine sulfated proteins. The VH and VL genes from one such sulfotyrosine-specific scFv were employed to generate a full length, sulfotyrosine-specific immunoglobulin.
1. An isolated sulfotyrosine antibody which is specific for a sulfated tyrosine antigenic determinant in a sulfotyrosine-containing polypeptide, and which comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO: 8, and a light chain variable region having the amino acid sequence of SEQ ID NO: 4.
2. The isolated sulfotyrosine antibody of claim 1 , which is a full length immunoglobulin selected from the classes consisting of IgA, IgD, IgE, IgG and IgM.
3. An isolated sulfotyrosine antibody fragment of the sulfotyrosine antibody of claim 1 or 2 , which is specific for a sulfated tyrosine determinant in a sulfotyrosine containing polypeptide.
4. The isolated sulfotyrosine antibody fragment of claim 3 , which is an scFv.
5. The isolated sulfotyrosine antibody fragment of claim 3 , which is an Fab or Fab′.
6. The isolated sulfotyrosine antibody of claim 1 or 2 , wherein the binding of the antibody to a sulfotyrosine antigen is inhibited by tyrosine sulfate in a competitive immunoassay.
7. The isolated sulfotyrosine antibody of claim 1 or 2 , wherein the antibody binds to a sulfotyrosine antigen at least two times more strongly than to an irrelevant antigen or antigen mixture in an ELISA.
8. The isolated sulfotyrosine antibody of claim 1 or 2 which is detectably labeled.
9. The isolated sulfotyrosine antibody of claim 1 , which is a recombinant antibody.
10. The isolated sulfotyrosine antibody of claim 2 , which is a recombinant antibody.
11. The isolated sulfotyrosine antibody of claim 9 or 10 which is detectably labeled.
12. A composition comprising the sulfotyrosine antibody of any one of claims 1 , 2 , 9 or 10 .
13. A composition comprising the sulfotyrosine antibody of claim 11 .