Simple Method for Repurification of Endotoxins for Biological Use

Appl Environ Microbiol. 2007 Mar;73(6):1803-8. doi: 10.1128/AEM.02452-06. Epub 2007 Jan 19.

Abstract

A method for obtaining highly purified endotoxin (lipopolysaccharide [LPS]) in a few hours by repurification of commercial or laboratory preparations was devised. It avoids the use of phenol, which is not suitable for phenol-soluble lipopolysaccharides nor for some industrial purposes. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization mass spectrometry analysis confirmed the integrity of the purified LPSs. The purified products did not activate Toll-like receptor 2 (TLR2), nuclear oligomerization domain 1 (NOD1), or NOD2 but did activate TLR4. Applied to different lipopolysaccharides, the method also improved their mass spectra, thus facilitating their structural analysis.

MeSH terms

  • Cell Line
  • Electrophoresis, Polyacrylamide Gel
  • Endotoxins / chemistry
  • Endotoxins / isolation & purification*
  • Humans
  • Immunologic Techniques*
  • Lipopolysaccharides / chemistry
  • Lipopolysaccharides / isolation & purification*
  • NF-kappa B / analysis
  • Nod1 Signaling Adaptor Protein / metabolism
  • Nod2 Signaling Adaptor Protein / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Toll-Like Receptor 2 / physiology
  • Toll-Like Receptor 4 / physiology

Substances

  • Endotoxins
  • Lipopolysaccharides
  • NF-kappa B
  • NOD1 protein, human
  • NOD2 protein, human
  • Nod1 Signaling Adaptor Protein
  • Nod2 Signaling Adaptor Protein
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4