Monoclonal antibodies against an identical short peptide sequence shared by two unrelated proteins.
Antipeptide antibodies provide the opportunity to explore the molecular basis for antigen-antibody recognition and to test theories of immune recognition. We investigated the possibility of raising monoclonal antipeptide antibodies against a specific epitope consisting of six amino acid residues, which is common to two unrelated proteins. The goal of this investigation was to analyze the reactivity of these epitope specific antibodies towards the same sequence in these two different proteins. A correlation between antibody reactivity and secondary structures of the same peptide sequence in different proteins could help to understand the ability of antipeptide antibodies to react with their cognate sequence in intact folded proteins. Monoclonal antibodies were raised against one hexamer sequence, PGTAPK, that is present in both thioredoxin and Fab New lambda-light chain. The antipeptide antibodies reacted only with thioredoxin but not with Fab New in ELISA's, immune precipitation and Western blots. Determination of the antibody specificity through binding tests with peptide analogs revealed the influence of the residue N-terminal from the hexamer epitope on antibody binding. Because of the observed influence of the N-1 adjacent residue in peptide analogs, the discrimination between the protein antigens could not be interpreted clearly as the result of the different hexamer conformations present in the native structures of the two proteins. However, analysis of the antibody reactivity with peptide analogs with varying "frame residues" surrounding the hexamer epitope indicates the possible discrimination of different peptide conformations by the antibody.
Schulze-Gahmen
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U
Research Institute of Scripps Clinic, La Jolla, CA 92037.
Wilson
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IA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis*; D006241:Haptens; D006825:Hybridomas / Q000276:immunology; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008805:Mice, Inbred A / Q000276:immunology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011487:Protein Conformation; D013879:Thioredoxins / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006241:Haptens; D007140:Immunoglobulin Fab Fragments; D009842:Oligopeptides; D010455:Peptides; D013879:Thioredoxins