Peptide vaccine against canine parvovirus: identification of two neutralization subsites in the N terminus of VP2 and optimization of the amino acid sequence.
The N-terminal domain of the major capsid protein VP2 of canine parvovirus was shown to be an excellent target for development of a synthetic peptide vaccine, but detailed information about number of epitopes, optimal length, sequence choice, and site of coupling to the carrier protein was lacking. Therefore, several overlapping peptides based on this N terminus were synthesized to establish conditions for optimal and reproducible induction of neutralizing antibodies in rabbits. The specificity and neutralizing ability of the antibody response for these peptides were determined. Within the N-terminal 23 residues of VP2, two subsites able to induce neutralizing antibodies and which overlapped by only two glycine residues at positions 10 and 11 could be discriminated. The shortest sequence sufficient for neutralization induction was nine residues. Peptides longer than 13 residues consistently induced neutralization, provided that their N termini were located between positions 1 and 11 of VP2. The orientation of the peptides at the carrier protein was also of importance, being more effective when coupled through the N terminus than through the C terminus to keyhole limpet hemocyanin. The results suggest that the presence of amino acid residues 2 to 21 (and probably 3 to 17) of VP2 in a single peptide is preferable for a synthetic peptide vaccine.
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D000917:Antibody Formation; D002213:Capsid / Q000276:immunology*; D036022:Capsid Proteins; D016384:Consensus Sequence; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay; D051379:Mice; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D017993:Parvovirus, Canine / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D011817:Rabbits / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship; D014765:Viral Vaccines / Q000276:immunology*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D010446:Peptide Fragments; D014765:Viral Vaccines