Preparation of refined and depolymerized capsular polysaccharide of Streptococcus pneumoniae
10.13200/j.cnki.cjb.004494
- VernacularTitle:肺炎链球菌荚膜精制多糖及降解多糖的制备
- Author:
LIU Yanli
- Publication Type:Journal Article
- Keywords:
Streptococcus pneumoniae;
Capsular polysaccharide(CPS);
Refined CPS;
Depolymerized CPS;
Acid precipitation;
Trifluoroacetic acid
- From:
Chinese Journal of Biologicals
2025;38(06):641-646+651
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prepare high-quality refined and depolymerized capsular polysaccharide(CPS) of Streptococcus pneumoniae for preparation of polysaccharide vaccine and polysaccharide conjugate vaccine against Streptococcus pneumoniae.Methods Refined CPS of type 6A was prepared by acid precipitation method from the inactivated fermentation broth of type 6A Streptococcus pneumoniae,and the final concentration of CaCl_2(80,160,240,320 mmol/L) and pH(2.5,3.0,3.5,4.0) were optimized.Three batches of type 6A refined CPSs were prepared by the optimized method,and the applicability of the method to polysaccharides of 5,7F,10A and 19F types was verified.After removing impurities from the inactivated broth of type 6 A Streptococcus pneumoniae by acid precipitation,the type 6 A depolymerized CPS was prepared by trifluoroacetic acid method,and the reaction temperature(8,25 and 40℃),the final concentration of trifluoroacetic acid(0.25,0.50,1.00 mol/L) and the reaction time(1,3,6 h) were optimized.Three batches of depolymerized CPSs were prepared by the optimized method,and compared with the high-pressure homogenization method,and then the applicability of trifluoroacetic acid method to depolymerized CPSs of 5,10A and 19F types was verified.The quality of refined and depolymerized CPSs was tested according to the methods specified in Chinese Pharmacopoeia(Vol Ⅲ,2020 edition).Results The optimum final concentration of CaCl_2 and pH of acid precipitation method were 160 mmol/L and 3.0 respectively.The prepared type 6A,5,7F,10A and 19F refined CPSs were all qualified.The optimum reaction temperature,reaction time and final concentration of trifluoroacetic acid were 25℃,3 h and 0.50 mol/L for trifluoroacetic acid method.The prepared type 6A,5,10A and 19F depolymerized CPSs were all qualified.Conclusion The refined and depolymerized CPSs of Streptococcus pneumoniae prepared in this study meet the requirements of Chinese Pharmacopoeia(Vol Ⅲ,2020 edition),and the preparation process is suitable for the preparation of various types of polysaccharides.
