Epitope mapping of bovine retinal S-antigen with monoclonal antibodies.
We examined the binding of seven murine monoclonal antibodies raised to S-antigen, an immunopathogenic, 404 residue photoreceptor cell autoantigen which induces experimental autoimmune uveoretinitis. S-antigen has also been identified as arrestin, a protein involved in the regulation of phototransduction. One additional monoclonal antibody (C10C10), raised to a synthetic peptide (peptide N) corresponding to residues 281 to 302 in bovine S-antigen, was also studied. In preliminary studies we examined the specificity of the antibody response to bovine S-antigen in sera from Balb/c mice. Western blots of mouse sera on the cyanogen bromide digest of bovine S-antigen demonstrated that all animals produced antibody which recognized epitopes within the C-terminal cyanogen bromide peptide designated CB46. Mice of the H-2d haplotype, including the Balb/c strain often used to produce monoclonal antibodies, showed little activity to cyanogen bromide peptides other than CB46. Also, all seven of the monoclonals raised to S-antigen are specific for epitopes in the CB46 peptide. The epitopes recognized by the monoclonal antibodies could be grouped into four distinct sites defined by peptides AE-1 (A2G5), peptide AA (PDS-1), peptide 19-OV (A9C6), and peptide 199 (BDS-1,2,3 and 4). The mono-clonal antibody, C10C10, raised to peptide N recognizes an epitope in the N peptide and binds to a larger cyanogen bromide peptide designated CB123 as well as intact S-antigen. Fine mapping of these epitopes was done with various subpeptides. None of the antibodies bound the known immunopathogenic peptide, peptide M, which resides in CB123 although the C10C10 antibody binds a peptide adjacent to peptide M.
Knospe
V
V
Department of Ophthalmology, University of Minnesota, Minneapolis.
Donoso
L A
LA
Banga
J P
JP
Yue
S
S
Kasp
E
E
Gregerson
D S
DS
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis*; D019393:Arrestin; D015153:Blotting, Western; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D005136:Eye Proteins / Q000032:analysis*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D010449:Peptide Mapping
D000911:Antibodies, Monoclonal; D000941:Antigens; D019393:Arrestin; D000939:Epitopes; D005136:Eye Proteins; D010446:Peptide Fragments