title: Cross-reactivity of antibodies to retro-inverso peptidomimetics with the parent protein histone H3 and chromatin core particle. Specificity and kinetic rate-constant measurements.
abstract: A series of monoclonal antibodies has been generated against an hexapeptide of sequence IRGERA corresponding to the C-terminal residues 130-135 of histone H3 and three analogues of this model peptide. The analogues correspond to the D-enatiomer, containing only D-residues, and two retro-peptides containing NH-CO bonds instead of natural amide peptide bonds. The chirality of each residue was maintained in the retro-peptide and inverted in the retro-inverso-peptide. Monoclonal antibodies were generated from mice immunized with the analogues coupled to neutral small unilamellar liposomes containing monophosphoryl lipid A as adjuvant. The reactivity of antibodies with the four analogues and with the parent protein H3 was studied in enzyme-linked immunosorbent assay and in a biosensor system. The equilibrium affinity constant (Ka) toward the retro-inverso-peptide of two out of three antibodies of IgG1 isotype induced against the L-hexapeptide was 7-75-fold higher than toward the homologous L-peptide. The range of Ka values of four antibodies of IgG1 and IgG2a isotypes generated against the retro-inverso-peptide was 0.6-1.9 x 10(9) M-1 for both the retro-inverso- and L-peptides. Furthermore, antibodies to the L- and retro-inverso-peptides cross-reacted strongly (in some cases better than with the homologous peptide) with the parent histone H3 and with chromatin subunits containing H3. The results are thus promising in respect to the potential use of retro-inverso-analogues, which are particularly stable, in the design of much more potent synthetic vaccines or to generate antibody probes.
authors.affiliation: Institute de Biologie Moléculaire et Cellulaire, UPR 9021 CNRS, Strasbourg, France.
authors.forename: N
authors.initials: N
authors.lastname: Benkirane
authors.forename: G
authors.initials: G
authors.lastname: Guichard
authors.forename: M H
authors.initials: MH
authors.lastname: Van Regenmortel
authors.forename: J P
authors.initials: JP
authors.lastname: Briand
authors.forename: S
authors.initials: S
authors.lastname: Muller
mesh_terms: D000276:Adjuvants, Immunologic; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002645:Chickens; D002843:Chromatin / Q000276:immunology*; D003429:Cross Reactions; D004912:Erythrocytes; D006657:Histones / Q000276:immunology*; D008050:Lipid A / Q000031:analogs & derivatives / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D011487:Protein Conformation
chemical_list: D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D002843:Chromatin; D006657:Histones; D008050:Lipid A; D010446:Peptide Fragments; C048436:monophosphoryl lipid A