PURIFICATION OF THE O ANTIGEN OF BACTEROIDES FRAGILIS SS. FRAGILIS NCTC 9343 FROM PHENOL-WATER EXTRACTS BY GEL FILTRATION AND CHROMATOGRAPHY ON DEAE-CELLULOSE AND HYDROXYLAPATITE

Abstract

The O antigen extracted from whole cells of B. fragilis ssp. fragilis NCTC 9343 with 45% aqueous phenol was purified by gel filtration and chromatography. First, the water phase was treated with RNase and DNase and passed through an agarose column. The chromatographic procedures included ion exchange on a column of DEAE-cellulose with a gradient of NaCl, and from the column of hydroxylapatite with 1 M phosphate buffer, pH 6.8. Inhibition of indirect hemagglutination [of sensitized sheep erythrocytes] was used to detect the O antigen in the eluates.