Separation of bacterial capsular and lipopolysaccharides by preparative electrophoresis

Abstract

Preparative gel electrophoresis was used to separate and purify extracellular, capsular and lipopolysaccharides (EPSs, CPSs, and LPSs, respectively) from crude bacterial extracts. The procedure effectively separates CPS from LPSs. In addition discreet size ranges of these various polysaccharides can be isolated. The ‘rough’ (R-type), ‘smooth’ (S-type), and ‘semi-smooth’ LPSs were separated from one another. In addition different size classes of ‘semi-smooth’, or S-type LPS, can be separated. This procedure was demonstrated for diverse bacterial species, including the soil bacteria Rhizobium fredii, and the enteric bacterial species, Salmonella enteritidis and Proteus mirabiis. In the latter case, it was also possible to separate capsular polysaccharide from its lipid-bound form.