Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Simplified purification method for Clostridium botulinum type E toxin.

J A Giménez1, H Sugiyama

  • 1Food Research Institute, University of Wisconsin, Madison 53706.

Applied and Environmental Microbiology
|December 1, 1987
PubMed
Summary

Researchers purified Clostridium botulinum type E toxin using advanced chromatography. This method yielded highly potent toxin, significantly exceeding previous reports in purity and toxicity.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Long-term follow-up of minimal residual disease in leukemia patients by monitoring WT1 (Wilms tumor gene) expression levels.

Blood·1996
Same author

Differential selectivity of M1 and M3 type muscarinic acetylcholine receptors in coupling with a G protein G01 alpha examined in Xenopus oocytes.

Neuroscience letters·1996
Same author

Catalytic activities of hammerhead ribozymes with a triterpenoid linker instead of stem/loop II.

FEBS letters·1996
Same author

Interleukin-6-mediated growth enhancement of cell lines derived from pyothorax-associated lymphoma.

Laboratory investigation; a journal of technical methods and pathology·1996
Same author

p70 S6 kinase sensitivity to rapamycin is eliminated by amino acid substitution of Thr229.

Journal of immunology (Baltimore, Md. : 1950)·1996
Same author

[Role of apoptosis in renal injury].

Nihon rinsho. Japanese journal of clinical medicine·1996

Area of Science:

  • Microbiology
  • Biochemistry
  • Protein Purification

Background:

  • Clostridium botulinum type E produces a potent neurotoxin.
  • Efficient purification of this toxin is crucial for research and potential therapeutic applications.

Purpose of the Study:

  • To develop and optimize a chromatography-based method for purifying Clostridium botulinum type E toxin.
  • To achieve high purity and yield of the toxin for further study.

Main Methods:

  • Three-step chromatography process involving DEAE-Sephadex, carboxymethyl Sepharose, and DEAE-Sephacel.
  • Toxin extraction from cells, concentration via precipitation, and buffer exchange.
  • Purification at 4°C on DEAE-Sephacel at a slightly alkaline pH.

Main Results:

  • A novel three-step chromatography protocol successfully purified Clostridium botulinum type E toxin.
  • The purified toxin (Mr, 145,000) was obtained in its bimolecular complex form.
  • Yields of at least 1.5 mg of pure toxin per liter of culture were achieved.
  • The toxicity of the purified toxin was 6 x 10^7 50% lethal doses per mg of protein, significantly higher than previously reported.

Conclusions:

  • The developed purification method is highly effective for obtaining pure and potent Clostridium botulinum type E toxin.
  • The achieved purity and toxicity levels represent a significant advancement in toxin purification.
  • This purified toxin is suitable for further research into its structure, function, and potential applications.

Related Experiment Videos