Dissection of an antibody paratope into peptides discloses the idiotope recognized by the cognate anti-idiotypic antibody.
Using methods of parallel synthesis, the complete amino acid sequence of an Ab 1 antibody (Tg 10, an anti-human thyroglobulin monoclonal antibody) was made in the form of a set of 100 synthetic overlapping peptides. This set of immobilized peptides was allowed to react with the cognate Ab2 (AI 10, a highly purified rabbit anti-idiotypic polyclonal antibody to Tg 10). A dominant peptide idiotope, INTFSGVPTYA, was thus mapped, which corresponds mainly to the CDR2 region from the V(H) domain of the Tg 10 mAb. A synthetic peptide replica of this idiotope was found to bind to AI 10 with an affinity (K(D) in the 10(-8) M range, as measured using BIACORE technology) which represents a significant part of the affinity of the complete Tg 10 antibody (K(D) in the 10(-9) M range). The synthetic peptide also elicited anti-idiotypic antibodies in rabbits that recognized specifically the Ab1 antibody in an Ab1- and antigen-inhibitable manner. The peptide idiotope was further characterized chemically by the identification of residues important for binding to the Ab2 and by modelization of its structure. Our approach makes it readily possible to map and characterize functional, continuous-type idiotopes that could be further used to manipulate the immune response by peptide technologies.
Laune
D
D
CNRS UMR 9921, Facult¿e de Pharmacie, 15 Avenue Charles Flahault, 34060 Cedex 2, Montpellier, France.
Molina
F
F
Mani
J
J
Del Rio
M
M
Bouanani
M
M
Pau
B
B
Granier
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D018604:Epitope Mapping; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D011817:Rabbits; D013954:Thyroglobulin / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D018985:Epitopes, B-Lymphocyte; D007140:Immunoglobulin Fab Fragments; D007135:Immunoglobulin Variable Region; D010455:Peptides; D013954:Thyroglobulin