19A型肺炎球菌多糖国家参考品的制备  

Preparation of national reference for serotype 19A pneumococcal polysaccharide

作　　者：陈琼[1] 王珊珊[1] 吕雪艳[2] 周学益 韩菲[3] 张亭[3] 叶强[1] CHEN Qiong;WANG Shan-shan;LV Xue-yan;ZHOU Xue-yi;HAN Fei;ZHANG Ting;YE Qiang(不详;National Institutes for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院,北京102629 [2]成都生物制品研究所有限责任公司,四川成都610023 [3]北京民海生物科技有限公司,北京102600

出　　处：《中国生物制品学杂志》2022年第4期433-436,共4页Chinese Journal of Biologicals

基　　金：国家重大新药创制专项(2018ZX09738006-006)。

摘　　要：目的 制备19A型肺炎球菌多糖国家参考品。方法 根据《中国药典》三部(2020版)对19A型肺炎球菌多糖候选参考品进行质量分析,包括蛋白质、核酸等杂质含量检测,氨基己糖和甲基戊糖等基团含量检测,以及分子大小分布等。以企业参考品为标定标准品,组织3个实验室采用速率比浊法对肺炎球菌多糖候选参考品进行协助标定。随机抽取50瓶候选参考品进行分装均匀性考察。由中国食品药品检定研究院通过速率比浊法对37、25和2~8℃放置一段时间的候选参考品进行热加速稳定性检测,并观察反复冻融3次对候选参考品19A型多糖含量的影响。结果 19A型肺炎球菌多糖候选参考品的各项质量指标均符合《中国药典》三部(2020版)标准;1H核磁共振结果确定候选参考品为19A型肺炎球菌多糖;协作标定结果显示,3个实验室测得的多糖含量RSD均小于5%,均值为565μg/mL。50瓶候选品多糖含量RSD为3.16%。2~8℃存放8周,25℃存放14 d,37℃存放3 d,以及反复冻融3次,稳定性均良好。结论 成功制备了19A型肺炎球菌多糖国家参考品,并通过协作标定确定了多糖含量,可用于肺炎球菌多糖疫苗中的多糖含量检测。Objective To prepare the national reference for serotype 19A pneumococcal polysaccharide. Methods According to the requirements in Chinese Pharmacopoeia(VolumeⅢ,2020 edition),the quality of serotype 19A pneumococcal polysaccharide candidate reference,including the contents of foreign matters such as protein and nucleic acid,the groups such as aminohexose and methyl pentose,as well as molecular size distribution,were analyzed. The pneumococcal polysaccharide candidate reference was collaboratively by rate nephelometry in three laboratories using the reference from manufacturer as standard. Fifty containers of candidate were randomly selected and investigated for the uniformity. The candidate was subjected to accelerant thermal stability test after storage at 37,25 or 2 ~ 8 ℃ for a certain time durations respectively by rate nephelometry by the National Institutes for Food and Drug Control(NIFDC).The effect of repeat freezing-thawing for 3 times on serotype 19A pneumococcal polysaccharide content of the candidate reference was observed. Results All the quality indexes of candidate reference met the requirements in Chinese Pharmacopoeia(Volume Ⅲ,2020 edition). 1H NMR confirmed the candidate as serotype 19A pneumococcal polysaccharide. The collaborative calibration results showed that the mean polysaccharide content determined by three laboratories was 565 μg/mL,of which the RSDs were less than 5%. The RSD of polysaccharide content of 50 containers of candidate was 3. 16%. The candidate showed high stability after storage at 2 ~ 8 ℃ for 8 weeks,at 25 ℃ for 14 d,at37 ℃ for 3 d,and after repeat freezing-thawing for 3 times. Conclusion The national reference for serotype 19A pneumococcal polysaccharide was successfully prepared,of which the polysaccharide content was determined by collaborative calibration. The reference may be used for determination of polysaccharide content in pneumococcal polysaccha-ride vaccine.

关 键 词：肺炎球菌多糖 国家参考品 速率比浊法 

分 类 号：R378.14[医药卫生—病原生物学] R392-33[医药卫生—基础医学]