Purification of Capsular Polysaccharide Complex from Gram-Negative Bacteria

Methods Mol Biol. 2019:1954:25-35. doi: 10.1007/978-1-4939-9154-9_3.

Abstract

Capsular polysaccharides are a dominant class of antigens from bacteria, both pathogenic and symbiotic or commensal. With the rise of awareness for the influence of the microbiota over immune system development and immune homeostasis, analysis of the antigens is more important than ever. Here we describe a method for the isolation of capsular polysaccharide from gram-negative bacteria, with the purification of polysaccharide from the commensal bacterium Bacteroides fragilis serving as an example. The method efficiently removes all detectable endotoxins and other lipid components, proteins, and nucleic acids, providing a source of capsular polysaccharide for immunologic study.

Keywords: Bacteria; Capsule; Carbohydrate; Endotoxin; Extraction; Glycan; Polysaccharide; Purification.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Capsules / chemistry*
  • Bacteroides Infections / microbiology
  • Bacteroides fragilis / chemistry*
  • Chemical Fractionation / instrumentation*
  • Chemical Fractionation / methods
  • Chemical Precipitation
  • Chromatography / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Equipment Design
  • Ethers / chemistry
  • Humans
  • Phenols / chemistry
  • Polysaccharides, Bacterial / chemistry
  • Polysaccharides, Bacterial / isolation & purification*

Substances

  • Ethers
  • Phenols
  • Polysaccharides, Bacterial