Endotoxin Removal from Escherichia coli Bacterial Lysate Using a Biphasic Liquid System

Methods Mol Biol. 2017:1600:107-112. doi: 10.1007/978-1-4939-6958-6_10.

Abstract

Lipopolysaccharide (LPS, endotoxin, pyrogen) which is a component of the outer membrane of most Gram-negative bacteria is a troubling contaminant of crude bacteriophage suspension. Therefore, its removal is important for bacteriophage applications especially in preparations dedicated for use in therapy with bacterial infections treatment. The method presented here is used for extractive removal of endotoxins from bacteriophage preparations with a water immiscible solvent such as 1-octanol. During extraction most of the phage lytic activity is retained in the aqueous phase, while endotoxin accumulates in the organic solvent. The levels of endotoxin in the aqueous bacteriophage rich fraction are determine by Limulus Amebocyte Lysate or EndoLISA assay and are extremely low.

Keywords: Bacteriophage; Extraction procedure; Lipopolysaccharides.

MeSH terms

  • Bacteriophages / metabolism
  • Escherichia coli / metabolism*
  • Lipopolysaccharides / chemistry*
  • Lipopolysaccharides / isolation & purification

Substances

  • Lipopolysaccharides