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Updated: Jun 9, 2026
Isolation and Preparation of Bacterial Cell Walls for Compositional Analysis by Ultra Performance Liquid Chromatography
Published on: January 15, 2014
A modified protocol for isolation and purity evaluation of a staphylococcal acidic polysaccharide by chromatography
Maria I Krevvata1, Nikos Afratis, Anastasia Spiliopoulou
1Laboratory of Microbiology, School of Medicine, University of Patras, Patras, Greece.
Abstract:
The extracellular slime of Staphylococcus epidermidis contains, amongst various macromolecules, an acidic polysaccharide (PS) of a molecular mass of 20 kDa with significant antigenic and biological properties. The isolation procedure used so far includes multiple fractionations in anion-exchange chromatographic columns before its final purification by gel filtration chromatography. This protocol is laborious, time-consuming and includes the risk of unnecessary loss of PS quantities. Because of the significance of this PS, a modified protocol resulting in an easier and quicker isolation procedure was developed. Furthermore, identification, purity, charge density and molecular integrity of the isolated polysaccharide were evaluated by a reverse-polarity capillary electrophoresis method.
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