黄热减毒活疫苗(鸡胚细胞)三级毒种库的建立和质量控制研究  

Establishment and quality control of virus seed bank of live attenuated yellow fever vaccine(chicken embryo cells)

作　　者：徐宏山[1] 黄艳秋 刘欣玉[1] 贾丽丽[1] 李玉华[1] XU Hong-shan;HUANG Yan-qiu;LIU Xin-yu;JIA Li-li;LI Yu-hua(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

出　　处：《药物分析杂志》2024年第10期1749-1755,共7页Chinese Journal of Pharmaceutical Analysis

基　　金：重大新药创制项目(2018ZX09738003-008)。

摘　　要：目的:筛选黄热减毒活疫苗17D-204原代鸡胚细胞适应株,制备三级毒种库,并对毒种库质量控制研究。方法:筛选黄热减毒活疫苗生产毒种17D-204株,在原代鸡胚细胞上适应和驯化培养,获得鸡胚细胞适应株,并对毒种库进行质量控制研究。结果:黄热减毒活疫苗17D-204在原代鸡胚细胞最适宜培养温度为37℃,最适宜感染复数(multiplicity of infection,MOI)为0.0048;建立了黄热减毒活疫苗(17D-204(CEC))原始种子、主代种子批、工作种子批三级毒种库。对毒种库鉴别试验、病毒滴度测定、无菌检查、支原体检查、外源病毒因子检查、分枝杆菌检查等项目研究,均符合《中华人民共和国药典》三部规定。高通量测序显示,17D-204(CEC)在原代鸡胚细胞上传代7代,其基因序列未发生改变。也未发现内、外源病毒因子污染。结论:筛选出了鸡胚细胞17D-204适应株17D-204(CEC),建立了三级毒种库,传代稳定性和遗传稳定性均良好,为原代鸡胚细胞取代鸡胚卵黄囊制备黄热减毒活疫苗奠定了坚实基础。Objective:To screen primary chicken embryo cells adapted strain of live attenuated yellow fever vaccine 17D-204,prepare virus seed banks and perform release tests.Methods:The virus seed of live attenuated yellow fever vaccine 17D-204 was adapted and domesticated on primary chicken embryo cells,and the chicken embryo cells adapted strain was obtained.Establish virus seed banks of live attenuated yellow fever vaccine(primary chicken embryo cells),and quality control was performed.Results:The optimum culture temperature of live attenuated yellow fever vaccine 17D-204 in primary chicken embryo cells was 37℃,and the optimum MOI(multiplicity of infection)was 0.0048.Three-level seed banks of primary seed bank,master seed bank,working seed bank of live attenuated yellow fever vaccine 17D-204(primary chicken embryo cells)[17D-204(CEC)]were established.The verification of virus titer,identification test,sterility test,mycoplasma,exogenous virus factor test,Mycobacterium test and other items were in line with the provisions of volumeⅢof the Chinese Pharmacopoeia.High throughput sequencing showed that there was no gene mutation and no internal and external virus factor pollution in the gene sequence of the virus seed banks.Conclusion:The primary chicken embryo cell adapted strain of live attenuated yellow fever vaccine 17D-204(CEC)can be screened out to establish the virus seed banks,with good passage stability and genetic stability.This study laid a firm foundation for the regeneration of live attenuated yellow fever vaccine from chicken yolk sac to chicken embryo cells.

关 键 词：黄热病毒 原代鸡胚细胞 毒种库 质量控制 黄热减毒活疫苗 17D-204 

分 类 号：R917[医药卫生—药物分析学]