Production of antibody against a synthetic peptide of Porphyromonas gingivalis 40-kDa outer membrane protein.
Porphyromonas gingivalis has been implicated as a major pathogen in periodontal diseases. We previously cloned a 40-kDa outer membrane protein (OMP) gene from P. gingivalis 381 and succeeded in producing sufficient quantities of the recombinant protein for experimental use. Since antibodies against the recombinant (r) 40-kDa OMP have potent ability to kill P. gingivalis cells by complement activation and opsonization, r40-kDa OMP has been the subject of considerable interest as a possible vaccine candidate. In this study, in order to develop a component vaccine, the immunodominant domain in 40-kDa OMP was identified. Peptides corresponding to portions of the N-terminal regions of 40-kDa OMP were synthesized chemically and their immunoreactivities with antibody against r40-kDa OMP were tested. The 16-mer peptide, LDDEYKERVFQTFVHY, was found to react strongly with the antibody. Furthermore, a rabbit antibody was prepared by immunization with the 16-mer peptide, cross-linked with dehydrofolate reductase, and its immunoreactivity was then examined. In a BIAcore experiment the antibody clearly reacted with r40-kDa OMP as well as P. gingivalis strains. These findings suggest that the 16-mer synthetic peptide may be useful for development of a component vaccine against P. gingivalis.
Tsurumi
Yoshiaki
Y
Department of Biochemistry, Nihon University School of Dentistry at Matsudo, Chiba 271-8587, Japan.
Hayakawa
Mitsuo
M
Shibata
Yasuko
Y
Abiko
Yoshimitsu
Y
D000276:Adjuvants, Immunologic; D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000942:Antigens, Bacterial / Q000032:analysis / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000276:immunology*; D001428:Bacterial Vaccines / Q000138:chemical synthesis; D004591:Electrophoresis, Polyacrylamide Gel; D007114:Immunization; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology; D016966:Porphyromonas gingivalis / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins
D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D010446:Peptide Fragments; D011994:Recombinant Proteins