5型肺炎球菌荚膜多糖纯化工艺的建立与优化  

Establishment and optimization of purification process for capsular polysaccharide of Streptococcus pneumoniae serotype 5

作　　者：曹欣[1,2] 徐永学 刘艳丽 高志鑫 宁云云 王元泽 王研研 郝倩 尹珊珊[1] 刘建凯[1,2] Cao Xin;Xu Yongxue;Liu Yanli;Gao Zhixin;Ning Yunyun;Wang Yuanze;Wang Yanyan;Hao Qian;Yin Shanshan;Liu Jiankai(Research&Development Center,Beijing Minhai Biotechnology Co.,Ltd.,Beijing 102600,China;Beijing New Combined Vaccine Engineering Research Center,Beijing 102600,China)

机构地区：[1]北京民海生物科技有限公司研发中心,北京102600 [2]北京市新型联合疫苗工程技术研究中心,北京102600

出　　处：《国际生物制品学杂志》2022年第4期200-204,共5页International Journal of Biologicals

基　　金：国家科技重大专项“重大新药创制”(2020ZX09301034)。

摘　　要：目的建立一种应用脱氧胆酸钠沉淀和离子交换色谱技术纯化5型肺炎球菌荚膜多糖的方法。方法 5型肺炎球菌发酵液经离心、超滤、脱氧胆酸钠沉淀处理后, 再经过Capto Q离子交换层析, 得到精制多糖原液。依据中国药典2020年版三部的相关制检规程对精制多糖原液进行检定分析。结果精制多糖原液中蛋白和核酸含量分别不超过3.0%和0.1%, 糖醛酸含量不低于18.4%, 氨基己糖含量不低于22.7%, 各项指标均符合中国药典标准;且多糖回收率达到60.0%以上。结论建立的5型肺炎球菌荚膜多糖纯化工艺可替代使用乙醇或苯酚的工艺, 且操作步骤简便, 具有很好的规模化应用前景。Objective To develop a method for purification of capsular polysaccharide from Streptococcus pneumoniae serotype 5 by protein precipitation with sodium deoxycholate(DOC)and ion exchange chromatography.Methods Streptococcus pneumoniae serotype 5 fermentation broth was treated by centrifugation,ultrafiltration and DOC precipitation,and then purified with Capto Q ion exchange chromatography to obtain the refined polysaccharide stock.The indexes of the purified polysaccharide bulk were analyzed according to the requirements of Chinese pharmacopoeia.Results The removal rates of protein and nucleic acid in fermentation broth were 97.0%and 99.9%.respectively.The contents of uronic acid and hexosamine in the purified polysaccharide were no less than18.4%and 22.7%.All the quality indicators meet the requirements of Chinese pharmacopoeia.The yield of polysaccharide was more than 60.0%.Conclusion An purification process of Streptococcus pneumoniae capsular polysaccharide is established,which can take place of methods using ethanol or phenol with good operability and a good prospect of industrial application.

关 键 词：肺炎球菌 荚膜多糖 脱氧胆酸钠 层析 纯化 

分 类 号：R392[医药卫生—免疫学]