A modified protocol for isolation and purity evaluation of a staphylococcal acidic polysaccharide by chromatography and capillary electrophoresis.
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.
Krevvata
Maria I
MI
Laboratory of Microbiology, School of Medicine, University of Patras, Patras, Greece.
Afratis
Nikos
N
Spiliopoulou
Anastasia
A
Malavaki
Christina J
CJ
Kolonitsiou
Fevronia
F
Anastassiou
Evangelos
E
Karamanos
Nikos K
NK
D019075:Electrophoresis, Capillary / Q000379:methods*; D005561:Formates / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D013212:Staphylococcus epidermidis / Q000737:chemistry*
D005561:Formates; D011135:Polysaccharides, Bacterial; C030544:formic acid