基质辅助激光解吸电离飞行时间质谱技术在13价肺炎球菌多糖蛋白结合疫苗生产用菌种质量控制中的作用  

Role of matrix-assisted laser desorption/ionization time of flight mass spectrometry for quality control of bacterial strain for production of 13-valent pneumococcus polysaccharide protein conjugate vaccine

作　　者：张丽芝 李阿妮 张新庄 程新 白贵杰 任克明 郑明睿 罗树权 ZHANG Li-zhi;Li A-ni;ZHANG Xin-zhuang;Cheng Xin;BAI Gui-jie;REN Ke-ming;ZHENG Ming-rui;LUO Shu-quan(First Department of Research,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第一研究室,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》2021年第12期1482-1487,共6页Chinese Journal of Biologicals

基　　金：国家科技重大专项(2018ZX09738-006)。

摘　　要：目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF MS)技术在13价肺炎球菌多糖蛋白结合疫苗生产用菌种质量控制中的作用。方法采用《中国药典》三部(2020版)中规定方法对13种血清型共24株肺炎球菌的经典表型进行鉴定,应用MALDI-TOF MS技术构建蛋白指纹图谱,根据质谱图的相似性进行聚类分析。同时采用MALDI-TOF MS技术检测不同荚膜丰度14型肺炎球菌(Streptococcus pneumoniae serotype 14,FL14)、不同来源FL5菌种及不同批次FL5工作菌种的蛋白指纹图谱的差异。结果13种血清型肺炎球菌均符合相关要求,蛋白指纹图谱存在差异,共分为3个聚类。不同荚膜丰度FL14的蛋白指纹图谱一致;FL5丹麦菌种及北京儿科医院菌种的指纹图谱有差异,且为不同聚类;不同批次FL5工作菌种的蛋白指纹谱图一致,均为同一个聚类。结论MALDI-TOF MS技术可作为经典细菌表型鉴定外的补充方法,丰富肺炎球菌菌种信息,可应用于疫苗用生产菌种的质量控制。Objective To investigate the role of matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS)in quality control of bacterial strain for production of 13-valent pneumococcus polysaccharide protein conjugate vaccine.Methods The classical phenotype of 24 strains of pneumococcus of 13 serotypes were identified by the method in Chinese Pharmacopoeia(VolumeⅢ,2020 edition),while the protein fingerprints were constructed by MALDI-TOF MS.Cluster analysis was performed according to the similarity of mass spectra.Meanwhile,the difference in protein fingerprints of Streptococcus pneumoniae serotype 14(FL14)with different capsular abundances,FL5 s trains from different origins and working seeds of FL5 of different batches were analyzed by MALDI-TOP MS.Results All the 13 serotypes of pneumococcus met the relevant requirements,while the protein fingerprints were different,which belonged to 3 clusters.The protein fingerprints of FL14 with different capsular abundances were in agreement,while those of FL5 from different origins were different.The FL5 strains from Denmark and from Beijing Children’s Hospital belonged to different clusters.However,the protein fingerprints of working strains of FL5 of different batches were in agreement,which belonged to one cluster.Conclusion MALDI-TOF MS may be used as a supplemental method for identification of classical bacterial phenotypes and provide more information on pneumococcus,thus may be used for the quality control of working strains for vaccine production.

关 键 词：13价肺炎多糖蛋白结合疫苗 肺炎球菌 基质辅助激光解吸电离飞行时间质谱 质量控制 

分 类 号：R378.1[医药卫生—病原生物学]