新型肺炎球菌荚膜多糖纯化方法的建立  

Development of a novel purification process for Streptococcus pneumonia capsular polysaccharide

作　　者：任克明 白贵杰 张丽芝 李阿妮 刘倩 张轶 王玺 张新庄 王剑虹 罗树权 沈荣 REN Ke-ming;BAI Gui-jie;ZHANG Li-zhi;LI A-ni;LIU Qian;ZHANG Yi;WANG Xi;ZHANG Xin-zhuang;WANG Jian-hong;LUO Shu-quan;SHEN Rong(Lanzhou Institute of Biological Products Co.,Ltd.,Center for Gansu Provincial Vaccine Engineering Research,Lanzhou 730046,Gansu Province,China)

机构地区：[1]兰州生物制品研究所有限责任公司甘肃省疫苗工程技术研究中心,甘肃兰州730046

出　　处：《中国生物制品学杂志》2020年第10期1181-1185,共5页Chinese Journal of Biologicals

基　　金：国家科技重大专项重大新药创新(2018ZX09738-006)。

摘　　要：目的建立一种无乙醇、无苯酚的新型肺炎球菌荚膜多糖纯化方法。方法以1型、6B型、14型肺炎球菌为模式样品,将发酵培养液依次用脱氧胆酸钠和核酸酶进行处理,以实现荚膜多糖与杂质的分离。再将此工艺应用于其他型别肺炎球菌发酵培养液纯化,获得纯化荚膜多糖。结果1型、6B型、14型肺炎球菌在纯化过程中核酸、蛋白的去除率均在99%以上,多糖回收率高于80%。应用此方法纯化的各型荚膜多糖各项指标均符合《欧洲药典》9.0版标准。结论该纯化方法适用于肺炎球菌荚膜多糖的规模化生产,且无血清型别限制,可替代传统的乙醇苯酚纯化工艺。Objective To develop a novel method for purification of Streptococcus pneumoniae capsular polysaccharide without alcohol and phenol.Methods S.pneumoniae types I,6 B and 14 were used as model samples.The fermentation broth was treated with sodium deoxycholate and nuclease successively in order to separate capsular polysaccharides from impurities.The process was applied to purification of fermentation broth of other serotypes of S.pneumoniae to obtain purified capsular polysaccharide.Results The removal rates of nucleic acid and protein during purification of S.pneumoniae types I,6 B and 14 were more than 99%,while the recovery rate of polysaccharide was more than 80%.All the quality indexes of capsular polysaccharide purified by this process met the requirements in European Pharmacopoeia 9.0.Conclusion Without serotype limitation,the developed purification process was suitable for large-scale production of S.pneumoniae capsular polysaccharide instead of the traditional purification process using ethanol.

关 键 词：肺炎球菌 荚膜多糖 纯化 乙醇 苯酚 

分 类 号：R378.14[医药卫生—病原生物学]