Premium
Epitope analysis using kinetic measurements of antibody binding to synthetic peptides presenting single amino acid substitutions
Author(s) -
ZederLutz Gabrielle,
Altschuh Danièle,
DeneryPapini Sandra,
Briand Jean Paul,
Van Regenmortel Marc H. V.,
Tribbick Gordon
Publication year - 1993
Publication title -
journal of molecular recognition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.401
H-Index - 79
eISSN - 1099-1352
pISSN - 0952-3499
DOI - 10.1002/jmr.300060205
Subject(s) - epitope , monoclonal antibody , chemistry , peptide , antibody , amino acid , dissociation constant , receptor–ligand kinetics , epitope mapping , biochemistry , peptide sequence , linear epitope , conformational epitope , kinetics , microbiology and biotechnology , biology , receptor , immunology , physics , quantum mechanics , gene
The fine modulation of peptide–antibody interactions was investigated with anti‐peptide monoclonal antibodies recognizing peptide 125–136 of the coat protein of tobacco mosaic virus. Nine synthetic peptides presenting single amino acid substitutions were selected for detailed analysis on the basis of their reactivity in ELISA. Kinetic measurements of the binding of four antibodies to these peptides performed with a biosensor instrument (BIAcore TM , Pharmacia) were used to quantify the contribution of individual residues to antibody binding. The results showed that even conservative exchanges of some residues in the epitope results in a small but significant decrease of the equilibrium affinity constant due mostly to a higher dissociation rate constant of the monoclonal antibodies. Two amino acid residues directly adjacent to the epitope, which appeared to play no role when tested by ELISA, were shown to influence the kinetics of binding. These data should be useful for computer modelling of the peptide–antibody interactions.