摘要
目的建立第二代肠道病毒71型(enterovirus 71,EV71)抗原国家标准品,用于体外效力(EV71抗原含量)质量控制。方法选取与第一代EV71抗原国家标准品相同来源的原料和冻干方法制备候选标准品(CS),经均匀性和稳定性考核合格后,分发给4个实验室,以EV71抗原国际标准品(WHO IS)和第一代EV71抗原国家标准品(NS)为标准进行协作标定及适用性研究。结果 CS与WHO IS及CS与NS均具有良好的平行性和线性。以WHO IS为标准时,CS检测的平均值为2 316 IU/mL,实验室内CV为4.3%~9.3%,实验室间CV为7.1%,95%置信区间为2 229~2 403 IU/mL;以NS为标准时,CS检测平均值为2 437 U/mL,实验室内CV为3.9%~10.7%,实验室间CV为5.3%,95%置信区间为2 355~2 519 U/mL。以CS为标准检测各企业生产的EV71收获液、原液和疫苗均具有较好的平行性和线性,且准确率均位于90%~110%之间。在2~8和37℃放置56 d,CS抗原含量稳定;CS复溶后置2~8℃保存7 d或-20℃反复冻融5次,抗原含量残留率均未超过90%~110%。结论建立了第二代EV71抗原国家标准品,赋值2 320 IU/mL(相当于2 440 U/mL)。
Objective To establish the second generation of national standard for enterovirus 71(EV71) antigen and use for the quality control of in vitro efficacy(EV71 antigen content).Methods The candidate standard(CS) was prepared by lyophilization with the materials from the same source as that for the first generation of national standard for EV71 antigen,and evaluated for homogeneity and stability.The qualified CS was distributed to four laboratories for collaborative calibration and studies on suitability using the international standard(WHO IS) and the first generation of national standard(NS) for EV71 antigen as standards.Results CS showed good parallelism and linea-rity with WHO IS and NS.Using WHO IS as a standard,the average value of CS was 2 316 IU/mL,while the intra-and inter-lab CVs were 4.3%~9.3% and 7.1% respectively,with a 95% confidence interval of 2 229 ~2 403 IU/mL.However,using NS as a standard,the average value of CS was 2 437 U/mL,while the CVs in intraand inter-assays were 3.9%~10.7% and 5.3% respectively,with a 95% confidence interval of 2 355~2 519 U/mL.The harvest,bulk and vaccine of EV71 manufactured by various manufacturers using CS as a standard showed good parallelism and linearity,while the accuracy was 90%~110%.The antigen content of CS showed no significant change after storage at 2~8 ℃ and 37 ℃ for 56 d.The residual antigen content of CS was between 90%~110% after reconstitution and storage at 2~8 ℃ for 7 d or after repeatedly freeze and thawing at-20 ℃ for 5 times.Conclusion The second generation of national standard for EV71 antigen was established and assigned as 2 320 IU/mL(equivalent to 2 440 U/mL).
作者
高帆
卞莲莲
吴星
么山山
王一平
毛群颖
梁争论
GAO Fan;BIAN Lian-lian;WU Xing;YAO Shan-shan;WANG Yi-ping;MAO Qun-ying;LIANG Zheng-lun(National Institutes for Food and Drug Control,Beijing 102629,China)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2021年第9期1069-1075,共7页
Chinese Journal of Biologicals
基金
国家科技重大专项(2018ZX09737-011)。
关键词
手足口病
肠道病毒71型
抗原含量
国家标准品
Hand
foot and mouth disease(HFMD)
Enterovirus 71(EV71)
Antigen content
National standard
