Optimization of removing protein in Haemophilus influenza type b capsular polysaccharide using Tween 80

To investigate the influence of Haemophilus influenzae type b (Hib) crude polysaccharide concentration,content of Tween 80 and (NH4)2SO4 on capsular polysaccharide recovery rate and residual content of protein,a single factor experiment and Box-Behnken experiment were conducted.The regression models were established and predicted the optimal parameters,of which the crude polysaccharide concentration was 8.54mg/mL,content of Tween 80 and (NH4)2SO4 were 12.9% and 28.12%,respectively,Furthermore,the experiments verified that the accuracy and conformity reached 93.9% and 90.3%.In this study,a new method by design of experiments (DOE) test was established and optimized to remove protein in Hib capsular polysaccharide,which could be applied in an actual production.