A simple and convenient method for the purification of Haemophilus Influenzae type b polysaccharide PRP

A simple and convenient method of removing the impurities from the broth like proteins, nucleic acids and endotoxins generated during the fermentation of Haemophilus Influenzae type b to recover pure polysaccharide (PRP) without using cationic detergent like cetyltrimethylammonium bromide (CTAB), enzymes and chromatographic purification methods. The present method removes the reagents added for the fermentation process by diafiltration and precipitation. The protein impurities are removed by sodium deoxycholate and sodium acetate with 30% ethanol precipitation, nucleic acid and endotoxins are removed by passing through depth and carbon filters while residual reagents by 300 kD membrane diafiltration. The purified PRP meets all the regulatory specifications and recovery is in the range of 60 to 80% with respect to the PRP produced during the fermentation process.