重组胰蛋白酶在腮腺炎减毒活疫苗和麻疹减毒活疫苗生产中的应用  

Application of recombinant trypsin in the production of live attenuated mumps vaccine and live attenuated measles vaccine

作　　者：许晚 张芹 汪张建 陈晓望 李贞妮 周锋 沈谊清 胡伊奇 徐晓婧 Xu Wan;Zhang Qin;Wang Zhangjian;Chen Xiaowang;Li Zhenni;Zhou Feng;Shen Yiqing;Hu Yiqi;Xu Xiaojing(Vaccine DivisionⅡ,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 201403,China;Producing Department,Shanghai Institute of Biological Products Co.,Ltd.,Shanghai 201403,China)

机构地区：[1]上海生物制品研究所有限责任公司疫苗二室,上海201403 [2]上海生物制品研究所有限责任公司生产部,上海201403

出　　处：《国际生物制品学杂志》2024年第4期207-211,共5页International Journal of Biologicals

基　　金：国家科技重大专项(2018ZX09738003-005)。

摘　　要：目的研究重组胰蛋白酶替代动物源胰蛋白酶用于腮腺炎减毒活疫苗和麻疹减毒活疫苗生产的可行性。方法根据动物源胰蛋白酶消化液活性计算所需重组胰蛋白酶浓度, 选择0.10~0.25 mg/ml的重组胰蛋白酶处理鸡胚组织生产2种疫苗, 以动物源胰蛋白酶为对照, 对细胞工厂活细胞总数、细胞贴壁率以及收获液的病毒滴度进行对比和统计学处理。结果和动物源胰蛋白酶组相比, 重组胰蛋白酶消化的鸡胚组织表观更细腻, 细胞易于分散。选择0.20 mg/ml重组胰蛋白酶用于腮腺炎和麻疹减毒活疫苗的生产, 每批鸡胚分散后的活细胞总数分别为1.19×10^(10)和1.63×10^(9), 对照组分别为9.31×10^(9)和1.27×10^(9);细胞贴壁率分别为63.01%和82.64%, 对照组分别为53.90%和75.99%;病毒收获液的病毒滴度均在正常范围内。结论 0.20 mg/ml的重组胰蛋白酶可替代动物源胰蛋白酶用于腮腺炎减毒活疫苗和麻疹减毒活疫苗的生产。Objective To evaluate the feasibility of recombinant trypsin as a substitute for animal-derived trypsin in production of live attenuated mumps and live attenuated measles vaccines.Methods The required concentration of recombinant trypsin was calculated based on the enzymatic activity of animal-derived trypsin and 0.10-0.25 mg/ml was selected to treat chicken embryo tissues and produce two vaccines.Animal-derived trypsin was used as control.The total number of viable cells,cell adherence rate,and virus titers of the virus harvests were compared and statistically processed.Results Compared with control group,the tissues of chicken embryos digested with recombinant trypsin appeared more delicate and the cells were easily dispersed.The total numbers of viable cells from each batch of chicken embryos after dispersion were 1.19×10^(10) and 1.63×10^(9) for 0.20 mg/ml recombinant trypsin groups in production of mumps and measles vaccines,9.31×10^(9) and 1.27×10^(9) for control group,respectively,while the cell adherence rates were 63.01%and 82.64%for 0.2 mg/ml groups,53.90%and 75.99%for control groups.Virus titers of the virus harvests were all within the normal range.Conclusion Recombinant trypsin at 0.20 mg/ml can be used as a substitute for animal-derived trypsin in the production of live attenuated mumps and measles vaccines.

关 键 词：腮腺炎疫苗 麻疹疫苗 重组胰蛋白酶 动物源胰蛋白酶 活细胞总数 细胞贴壁率 滴度/病毒 

分 类 号：R392-33[医药卫生—免疫学]