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J. Biol. Chem., Vol. 277, Issue 23, 20468-20476, June 7, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/23/20468    most recent M200387200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Brett, P. J. Articles by Charalambous, B. M. Search for Related Content PubMed PubMed Citation Articles by Brett, P. J. Articles by Charalambous, B. M. Social Bookmarking                      What's this?

Characterization of Oligopeptides That Cross-react with Carbohydrate-specific Antibodies by Real Time Kinetics, In-solution Competition Enzyme-linked Immunosorbent Assay, and Immunological Analyses^*

Paul J. Brett, Harmale Tiwana, Ian M. Feavers^§, and Bambos M. Charalambous^¶

From the  Department of Medical Microbiology, Royal Free and University College Medical School, Royal Free Campus, University College London, Rowland Hill Street, London NW3 2PF and ^§ Division of Bacteriology, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Hertfordshire EN6 3QG, United Kingdom

Phage displaying random cyclic 7-mer, and linear 7-mer and 12-mer peptides at the N terminus of the coat protein, pIII, were panned with the murine monoclonal antibody, 9-2-L379 specific for meningococcal lipo-oligosaccharide. Five cyclic peptides with two sequence motifs, six linear 7-mers, and five linear 12-mers with different sequence motifs were identified. Only phage displaying cyclic peptides were specifically captured by and were antigenic for 9-2-L379. Monoclonal antibody 9-2-L379 exhibited "apparent" binding affinities to the cyclic peptides between 11 and 184 nM, comparable with lipo-oligosaccharide. All cyclic peptides competed with the binding of 9-2-L379 to lipo-oligosaccharide with EC₅₀ values in the range 10-105 µM, which correlated with their apparent binding affinities. Structural modifications of the cyclic peptides eliminated their ability to bind and compete with monoclonal antibody 9-2-L379. Mice (C3H/HeN) immunized with the cyclic peptide with optimal apparent binding affinity and EC₅₀ of competition elicited cross-reactive antibodies to meningococcal lipo-oligosaccharide with end point dilution serum antibody titers of 3200. Cyclic peptides were converted to T-cell-dependent immunogens without disrupting these properties by C-terminal biotinylation and complexing with NeutrAvidin®. The data indicate that constrained peptides can cross-react with a carbohydrate-specific antibody with greater specificity than linear peptides, and critical to this specificity is their structural conformation.

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