Epitope analysis using kinetic measurements of antibody binding to synthetic peptides presenting single amino acid substitutions.
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, 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 resulted 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.
Zeder-Lutz
G
G
Laboratoire d'Immunochimie, Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France.
Altschuh
D
D
Denery-Papini
S
S
Briand
J P
JP
Tribbick
G
G
Van Regenmortel
M H
MH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D001666:Binding Sites, Antibody; D015374:Biosensing Techniques; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000276:immunology / Q000378:metabolism*; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship; D014027:Tobacco Mosaic Virus; D014764:Viral Proteins / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D009842:Oligopeptides; D014764:Viral Proteins