Production of functional ScFv inhibiting Streptococcus mutans glucosyltransferase activity from a hybridoma P126

Hiroyoshi Tagawa, Mitsuo Hayakawa, Yasuko Shibata, Kazuo Fukushima, Sheng Yang Lee, Teruaki Shiroza, Yoshimitsu Abiko

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Streptococcus mutans has been considered the principal etiologic agent of dental caries in humans. The glucosyltransferase-I (GTF-I), which synthesized adhesive water-insoluble glucans from sucrose, has been demonstrated to be an important cariogenic property. Water-insoluble glucans (WIG) synthesized by S. mutans GTF-I can mediate sucrose-enhanced colonization on tooth surfaces and form dental plaque. It has been suggested that inhibition of WIG synthesis decreases bacterial colonization and cariogenicity. Indeed, the use of GTF enzymes as a vaccine antigen resulted in protection from experimental dental caries in rodents. However, it is preferable to eliminate unwanted immune response during active immunization of humans. To prevent this incidence, we attempted to produce the single-chain variable fragment (ScFv) antibody against GTF-I to develop passive immunization for dental caries. Hybridomas producing monoclonal antibody (MAb) that inhibited WIG synthesis by GTF-I have been constructed. Using mRNA from an IgG1 hybridoma P126, cDNAs encoding the variable fragments of the L and H chains of IgG1 from the hybridoma P126 were cloned by RT-PCR-based techniques and then transformed into an Escherichia coli expression system. The purified ScFv antibody recognized the recombinant (r) GTF-I proteins and was capable of inhibiting the WIG synthesis of rGTF-I.

Original languageEnglish
Pages (from-to)305-310
Number of pages6
JournalHybridoma and Hybridomics
Volume23
Issue number5
DOIs
Publication statusPublished - Oct 2004

Fingerprint

Glucosyltransferases
Streptococcus mutans
Glucans
Hybridomas
Single-Chain Antibodies
Dental Caries
Water
Sucrose
Immunoglobulin G
Dental Plaque
Passive Immunization
Adhesives
Rodentia
Tooth
Vaccination
Vaccines
Complementary DNA
Monoclonal Antibodies
glucosyltransferase I
Escherichia coli

ASJC Scopus subject areas

  • Genetics
  • Immunology

Cite this

Production of functional ScFv inhibiting Streptococcus mutans glucosyltransferase activity from a hybridoma P126. / Tagawa, Hiroyoshi; Hayakawa, Mitsuo; Shibata, Yasuko; Fukushima, Kazuo; Lee, Sheng Yang; Shiroza, Teruaki; Abiko, Yoshimitsu.

In: Hybridoma and Hybridomics, Vol. 23, No. 5, 10.2004, p. 305-310.

Research output: Contribution to journalArticle

Tagawa, Hiroyoshi ; Hayakawa, Mitsuo ; Shibata, Yasuko ; Fukushima, Kazuo ; Lee, Sheng Yang ; Shiroza, Teruaki ; Abiko, Yoshimitsu. / Production of functional ScFv inhibiting Streptococcus mutans glucosyltransferase activity from a hybridoma P126. In: Hybridoma and Hybridomics. 2004 ; Vol. 23, No. 5. pp. 305-310.
@article{8e3b323a4ee7408ab3485bb3ba289609,
title = "Production of functional ScFv inhibiting Streptococcus mutans glucosyltransferase activity from a hybridoma P126",
abstract = "Streptococcus mutans has been considered the principal etiologic agent of dental caries in humans. The glucosyltransferase-I (GTF-I), which synthesized adhesive water-insoluble glucans from sucrose, has been demonstrated to be an important cariogenic property. Water-insoluble glucans (WIG) synthesized by S. mutans GTF-I can mediate sucrose-enhanced colonization on tooth surfaces and form dental plaque. It has been suggested that inhibition of WIG synthesis decreases bacterial colonization and cariogenicity. Indeed, the use of GTF enzymes as a vaccine antigen resulted in protection from experimental dental caries in rodents. However, it is preferable to eliminate unwanted immune response during active immunization of humans. To prevent this incidence, we attempted to produce the single-chain variable fragment (ScFv) antibody against GTF-I to develop passive immunization for dental caries. Hybridomas producing monoclonal antibody (MAb) that inhibited WIG synthesis by GTF-I have been constructed. Using mRNA from an IgG1 hybridoma P126, cDNAs encoding the variable fragments of the L and H chains of IgG1 from the hybridoma P126 were cloned by RT-PCR-based techniques and then transformed into an Escherichia coli expression system. The purified ScFv antibody recognized the recombinant (r) GTF-I proteins and was capable of inhibiting the WIG synthesis of rGTF-I.",
author = "Hiroyoshi Tagawa and Mitsuo Hayakawa and Yasuko Shibata and Kazuo Fukushima and Lee, {Sheng Yang} and Teruaki Shiroza and Yoshimitsu Abiko",
year = "2004",
month = "10",
doi = "10.1089/hyb.2004.23.305",
language = "English",
volume = "23",
pages = "305--310",
journal = "Monoclonal Antibodies in Immunodiagnosis and Immunotherapy",
issn = "2167-9436",
publisher = "Mary Ann Liebert Inc.",
number = "5",

}

TY - JOUR

T1 - Production of functional ScFv inhibiting Streptococcus mutans glucosyltransferase activity from a hybridoma P126

AU - Tagawa, Hiroyoshi

AU - Hayakawa, Mitsuo

AU - Shibata, Yasuko

AU - Fukushima, Kazuo

AU - Lee, Sheng Yang

AU - Shiroza, Teruaki

AU - Abiko, Yoshimitsu

PY - 2004/10

Y1 - 2004/10

N2 - Streptococcus mutans has been considered the principal etiologic agent of dental caries in humans. The glucosyltransferase-I (GTF-I), which synthesized adhesive water-insoluble glucans from sucrose, has been demonstrated to be an important cariogenic property. Water-insoluble glucans (WIG) synthesized by S. mutans GTF-I can mediate sucrose-enhanced colonization on tooth surfaces and form dental plaque. It has been suggested that inhibition of WIG synthesis decreases bacterial colonization and cariogenicity. Indeed, the use of GTF enzymes as a vaccine antigen resulted in protection from experimental dental caries in rodents. However, it is preferable to eliminate unwanted immune response during active immunization of humans. To prevent this incidence, we attempted to produce the single-chain variable fragment (ScFv) antibody against GTF-I to develop passive immunization for dental caries. Hybridomas producing monoclonal antibody (MAb) that inhibited WIG synthesis by GTF-I have been constructed. Using mRNA from an IgG1 hybridoma P126, cDNAs encoding the variable fragments of the L and H chains of IgG1 from the hybridoma P126 were cloned by RT-PCR-based techniques and then transformed into an Escherichia coli expression system. The purified ScFv antibody recognized the recombinant (r) GTF-I proteins and was capable of inhibiting the WIG synthesis of rGTF-I.

AB - Streptococcus mutans has been considered the principal etiologic agent of dental caries in humans. The glucosyltransferase-I (GTF-I), which synthesized adhesive water-insoluble glucans from sucrose, has been demonstrated to be an important cariogenic property. Water-insoluble glucans (WIG) synthesized by S. mutans GTF-I can mediate sucrose-enhanced colonization on tooth surfaces and form dental plaque. It has been suggested that inhibition of WIG synthesis decreases bacterial colonization and cariogenicity. Indeed, the use of GTF enzymes as a vaccine antigen resulted in protection from experimental dental caries in rodents. However, it is preferable to eliminate unwanted immune response during active immunization of humans. To prevent this incidence, we attempted to produce the single-chain variable fragment (ScFv) antibody against GTF-I to develop passive immunization for dental caries. Hybridomas producing monoclonal antibody (MAb) that inhibited WIG synthesis by GTF-I have been constructed. Using mRNA from an IgG1 hybridoma P126, cDNAs encoding the variable fragments of the L and H chains of IgG1 from the hybridoma P126 were cloned by RT-PCR-based techniques and then transformed into an Escherichia coli expression system. The purified ScFv antibody recognized the recombinant (r) GTF-I proteins and was capable of inhibiting the WIG synthesis of rGTF-I.

UR - http://www.scopus.com/inward/record.url?scp=7244236290&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=7244236290&partnerID=8YFLogxK

U2 - 10.1089/hyb.2004.23.305

DO - 10.1089/hyb.2004.23.305

M3 - Article

C2 - 15672609

AN - SCOPUS:7244236290

VL - 23

SP - 305

EP - 310

JO - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

JF - Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

SN - 2167-9436

IS - 5

ER -