Selection and characterization of cyclic peptides that bind to a monoclonal antibody against meningococcal L3,7,9 lipopolysaccharides.
There is still no general vaccine for prevention of disease caused by group-B meningococcal strains. Meningococcal lipopolysaccharides (LPSs) have received attention as potential vaccine candidates, but concerns regarding their safety have been raised. Peptide mimics of LPS epitopes may represent safe alternatives to immunization with LPS. The monoclonal antibody (MoAb) 9-2-L3,7,9 specific for Neisseria meningitidis LPS immunotype L3,7,9 is bactericidal and does not cross-react with human tissue. To explore the possibility of isolating peptide mimics of the epitope recognized by MoAb 9-2-L3,7,9, we have constructed two phage display libraries of six and nine random amino acids flanked by cysteines. Furthermore, we developed a system for the easy exchange of peptide-encoding sequences from the phage-display system to a hepatitis B core (HBc) expression system. Cyclic peptides that specifically bound MoAb 9-2-L3,7,9 at a site overlapping with the LPS-binding site were selected from both libraries. Three out of four tested peptides which reacted with MoAb 9-2-L3,7,9 were successfully presented as fusions to the immunodominant loop of HBc particles expressed in Escherichia coli. However, both peptide conjugates to keyhole limpet haemocyanin and HBc particle fusions failed to give an anti-LPS response in mice.
Lauvrak
V
V
Department of Molecular Biosciences, University of Oslo, Norway. vigdis.lauvrak@bio.uio.no
Berntzen
G
G
Heggelund
U
U
Herstad
T K
TK
Sandin
R H
RH
Dalseg
R
R
Rosenqvist
E
E
Sandlie
I
I
Michaelsen
T E
TE
D000818:Animals; D000907:Antibodies, Bacterial / Q000737:chemistry / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D001665:Binding Sites / Q000276:immunology; D006433:Hemocyanins / Q000276:immunology; D008070:Lipopolysaccharides / Q000276:immunology*; D051379:Mice; D009345:Neisseria meningitidis / Q000276:immunology*; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000737:chemistry / Q000276:immunology*; D011485:Protein Binding
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides; D019151:Peptide Library; D010456:Peptides, Cyclic; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin