title: Mapping epitopes of neutralizing monoclonal antibodies using phage random peptide libraries.
abstract: Identification of protective determinants from microbial proteins is a necessary step in the rational design of subunit vaccines. We have previously used a synthetic peptide scan (Pepscan) assay to map a panel of eight neutralizing monoclonal antibodies (mAb; designated as C1.1 to C1.8) to a common motif sequence from Chlamydia trachomatis. In the present study, five of the eight mAbs were used to screen phage random peptide libraries. mAbs C1.1 and C1.3 selected a motif sequence of G-L-X-N-D from a pIII-based phage random peptide library and a motif sequence of G-X-X-N-D from a pVIII-based random peptide library while mAbs C1.6 to C1.8 failed to select recognizable motifs from either of the phage libraries. However, C1.6 to C1.8 bound to the same motif sequence displayed on phage when the appropriate conformational constraints were imposed onto the motif sequence. Thus the specificity of the mAbs identified on Pepscan assays correlates with the mAbs' dependence on local epitope constraints displayed on the phage surface.
authors.affiliation: Department of Medical Microbiology, University of Manitoba, Winnipeg, Canada.
authors.forename: G
authors.initials: G
authors.lastname: Zhong
authors.forename: J D
authors.initials: JD
authors.lastname: Berry
authors.forename: S
authors.initials: S
authors.lastname: Choukri
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D001435:Bacteriophages / Q000235:genetics; D002692:Chlamydia trachomatis / Q000276:immunology*; D018604:Epitope Mapping; D015723:Gene Library; D008969:Molecular Sequence Data
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins