title: Equilibrium competitive inhibition analysis of synthetic peptide antigens from myelin basic protein as affected by the dual-dilution phenomenon.
abstract: It was shown that 125I-labelled and unlabelled forms of synthetic encephalitogenic peptide S82 (residues 65-83 plus glycine) of bovine myelin basic protein (MBP-Bov) were equally competitive in dual-dilution radioimmunoassays with rat- and rabbit-anti-S82 antisera without causing much deviation even at the extremes of the dual-dilution binding curves (solved in terms of total S82). With other antisera the deviations caused by the addition of unlabelled S82 were much greater than would be expected among repetitive assays with labelled antigen alone, and the excessive deviations were usually more prominent in one region of the dual-dilution binding curve than in another. Thus, establishing equivalence between labelled and unlabelled antigen with respect to one antiserum even at several dilutions does not establish proportionate sharing with respect to all antisera at all antigen concentrations. A method of dual-dilution equilibrium competitive inhibition analysis was devised that took this precaution into account. By means of the method, synthetic MBP-Bov peptides representing different parts of the S82 sequence were compared with homologous S82 peptide for their inhibitory effects upon dually diluted [125I]S82-anti-S82 systems. By this process several different S82 determinants were pinpointed, some with high affinity antibodies, others with low affinity antibodies, yet others equally well at high or low affinity.
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis; D000915:Antibody Affinity; D000941:Antigens / Q000032:analysis*; D001667:Binding, Competitive; D004676:Myelin Basic Protein / Q000032:analysis / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology*; D011817:Rabbits; D011863:Radioimmunoassay / Q000379:methods*; D051381:Rats; D011917:Rats, Inbred Lew
chemical_list: D000906:Antibodies; D000941:Antigens; D004676:Myelin Basic Protein; D010446:Peptide Fragments