Purification of LPS from Leptospira

Methods Mol Biol. 2020:2134:53-65. doi: 10.1007/978-1-0716-0459-5_6.

Abstract

Leptospira species are one of the few spirochetes to possess a lipopolysaccharide (LPS) embedded in their outer membrane. Two protocols are currently available to extract and/or purify the leptospiral lipopolysaccharides: the rapid proteinase K method and the classical hot water/phenol extraction. The first method allows to get a quick overview of the LPS O antigen structure, whereas the second method is fitted to study the immunological properties of the leptospiral LPS. These two methods will be detailed in this chapter. Methodologies to assess the quality of the purification, such as the modified silver staining coloration, will also be reviewed. Both advantages and limitations of the different analyses will be described.

Keywords: Hot water/phenol extraction; Leptospira ssp.; Lipopolysaccharide/endotoxin purification; Proteinase K; Silver staining.

MeSH terms

  • Clinical Laboratory Techniques / methods*
  • Endopeptidase K / chemistry
  • Leptospira / chemistry*
  • Lipopolysaccharides / chemistry*
  • Lipopolysaccharides / isolation & purification*
  • O Antigens / chemistry
  • O Antigens / isolation & purification
  • Phenols / chemistry
  • Silver Staining / methods

Substances

  • Lipopolysaccharides
  • O Antigens
  • Phenols
  • Endopeptidase K