Expression of Hc fragment from Clostridium botulinum neurotoxin serotype B in Escherichia coli and its use as a good immunogen

Hum Vaccin. 2010 Jun;6(6):462-6. doi: 10.4161/hv.6.6.11709. Epub 2010 Jun 1.

Abstract

A good immunogen was developed as an effective recombinant vaccine candidate that protected mice against botulinum neurotoxin serotype B (BoNT/B) intoxication. The Hc fragment of Clostridium botulinum neurotoxin type B (rBoNT/B-Hc) was cloned into the bacterial expression vector pQE-30, and the resulting vector was successfully expressed in the Escherichia coli M15 strain. The purified rBoNT/B-Hc protein was used to vaccinate mice and evaluate their survival against challenge with native BoNT/B. The mice that received three subcutaneous injections of rBoNT/B-Hc immunogen doses ranging from 0.25 to 6.25 μg mixed with Freund's adjuvant were completely protected against an intraperitoneal (i.p.) administration of 10,000 50% lethal doses (LD50) of BoNT/B. A dose response was observed in both the ELISA antibody titers and protective efficacy with increasing dose of immunogen. The work presented here demonstrates that the purified rBoNT/B-Hc was a highly effective immunogen and able to protect against a high-dose neurotoxin challenge.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Bacterial Vaccines / administration & dosage
  • Bacterial Vaccines / genetics
  • Bacterial Vaccines / immunology*
  • Botulinum Toxins / administration & dosage
  • Botulinum Toxins / genetics
  • Botulinum Toxins / immunology*
  • Botulinum Toxins, Type A
  • Botulism / blood
  • Botulism / immunology
  • Botulism / prevention & control*
  • Clostridium botulinum type B / immunology*
  • Dose-Response Relationship, Immunologic
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Female
  • Injections, Subcutaneous
  • Mice
  • Mice, Inbred BALB C
  • Peptide Fragments / administration & dosage
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology
  • Protein Engineering
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Vaccination
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology

Substances

  • Antibodies, Bacterial
  • Bacterial Vaccines
  • Peptide Fragments
  • Recombinant Proteins
  • Vaccines, Synthetic
  • rimabotulinumtoxinB
  • Botulinum Toxins
  • Botulinum Toxins, Type A