Production of a single-chain variable fraction capable of inhibiting the Streptococcus mutans glucosyltransferase in Bacillus brevis: construction of a chimeric shuttle plasmid secreting its gene product.
Periodontitis and dental caries are common oral diseases, in these days, and the passive immunization is one of the most effective approaches for prevention. For this purpose, we have constructed mouse and human monoclonal antibodies to inhibit the Porphyromonas gingivalis-associated hemagglutination and coaggregation. In addition, an artificial antibody, single-chain variable fraction, or scFv, which also inhibited the hemagglutination, was constructed. Specifically for dental caries, mouse and human monoclonal antibodies that inhibited the glucosyltransferase (GTF) activity, responsible for biofilm formation, were also constructed. The advantage of scFv over the native antibody is that the former molecule does not induce possible side-effects due to Fc, such as autoimmune disease, because it consists only of variable regions originating from both heavy and light chains. To increase the abilities of the antibody preparations, we attempted to construct an additional scFv using Bacillus brevis, a secretion-proficient gram-positive bacterium, as a host cell. An scFv protein possessing the same biological activity as that of the parental antibody was successfully secreted from a B. brevis transformant following the construction of a chimeric shuttle plasmid, which was accomplished by employing a new heterodimer system.
Shiroza
Teruaki
T
Department of Biochemistry, Nihon University School of Dentistry at Matsudo, Matsudo, 271-8587, Chiba, Japan. tshiroza@mascat.nihon-u.ac.jp <tshiroza@mascat.nihon-u.ac.jp>
Shinozaki-Kuwahara
Noriko
N
Hayakawa
Mitsuo
M
Shibata
Yasuko
Y
Hashizume
Tomomi
T
Fukushima
Kazuo
K
Udaka
Shigezo
S
Abiko
Yoshimitsu
Y
D018528:ATP-Binding Cassette Transporters / Q000235:genetics; D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D019677:Artificial Gene Fusion; D001407:Bacillus / Q000235:genetics*; D001422:Bacterial Adhesion; D001426:Bacterial Proteins; D001483:Base Sequence; D004274:DNA, Recombinant / Q000235:genetics; D005936:Glucans / Q000096:biosynthesis; D005964:Glucosyltransferases / Q000276:immunology* / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013295:Streptococcus mutans / Q000201:enzymology* / Q000276:immunology / Q000472:pathogenicity; D014169:Transformation, Bacterial
D018528:ATP-Binding Cassette Transporters; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D004274:DNA, Recombinant; D005936:Glucans; D007135:Immunoglobulin Variable Region; D011993:Recombinant Fusion Proteins; D005964:Glucosyltransferases