Expression of Hc fragment from Clostridium botulinum neurotoxin serotype B in Escherichia coli and its use as a good immunogen.
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.
Laboratory of Toxinology, State key Laboratory of Pathogen and Biosecurity, The institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing China.
Yan-Li
YL
Gao
Shan
S
Gao
Lin
L
Kang
Chong
C
Nie
Jing-Lin
JL
Wang
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001905:Botulinum Toxins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D019274:Botulinum Toxins, Type A; D001906:Botulism / Q000097:blood / Q000276:immunology / Q000517:prevention & control*; D047014:Clostridium botulinum type B / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D007279:Injections, Subcutaneous; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
D000907:Antibodies, Bacterial; D001428:Bacterial Vaccines; D010446:Peptide Fragments; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C096323:rimabotulinumtoxinB; D001905:Botulinum Toxins; D019274:Botulinum Toxins, Type A