Fine specificity of antibody recognition of carcinoma-associated epithelial mucins: antibody binding to synthetic peptide epitopes.
The protein core of polymorphic epithelial mucins consists predominantly of a repeating 20 amino acid peptide motif. Many monoclonal antibodies reactive with breast carcinomas recognise determinants located within the mucin protein core, and epitope mapping techniques have demonstrated that these antibodies bind to epitopes of three, four or five amino acids within the hydrophilic sequence, P D T R P A P. Each of these mucin core-reactive antibodies map to epitopes containing the central arginine residue. The fine specificity of a panel of anti-mucin antibodies binding to the tetrameric peptides P D T R or R P A P (synthesised on the heads of polyethylene pins) was examined by systematically replacing each amino acid in turn with all other 19 natural amino acids, and then testing these analogues for antibody binding. We have (i) identified those amino acids in epitopes which are essential for antibody binding, (ii) shown that for each epitope there is a hierarchy of residues required for immune recognition--certain amino acids may be replaced with little or no loss of antibody binding, while the presence of others is essential, and (iii) concluded that antibody specificity is further regulated by the residue(s) flanking an epitope motif which may impose conformational constraints upon the presentation of the epitope to an antibody.
Briggs
S
S
Cancer Research Campaign Laboratories, University of Nottingham, U.K.
Price
M R
MR
Tendler
S J
SJ
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D000951:Antigens, Neoplasm / Q000737:chemistry*; D000939:Epitopes / Q000737:chemistry*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry / Q000276:immunology*; D010449:Peptide Mapping
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000939:Epitopes; D009077:Mucins