冻干乙型脑炎减毒活疫苗的冻干工艺优化研究

Studies on the optimized freeze drying process for lyophilized live Japanese encephalitis vaccine

为了研究优化冻干乙型脑炎减毒活疫苗的冻干工艺,运用正交试验法L934考察不同冻干参数对该疫苗成品质量的影响,以外观、残余水分、病毒滴度以及热稳定性为直观分析指标,并对病毒滴度进行方差分析。结果显示,主干燥时间和主干燥真空压力对病毒滴度的影响有显著统计学意义(P<0.05)。优化的冻干参数为预冻温度-35℃、时间2h;主干燥温度从-35℃升温至-10℃,再由-10℃升温至33℃,总耗时16h,真空压力0.220mbar;二次干燥的温度维持于30℃,真空压力为0.001mbar,终点测试压力无变化时结束。对冻干乙型脑炎减毒活疫苗冻干参数筛选优化后,可得到较佳的冻干曲线,经验证该曲线适用于生产。

To study the optimized freeze drying strategy of lyophilized live Japanese encephalitis vaccine（JEV ）. The parameters of lyophilization process were optimized by means of orthogonal experimental design L934. The qualities of lyophilized live JEV were tested by shape, residual water, virus titer and titer after heat exposure. In addition, the virus titer was analyzed by analysis of variance. The results showed that the effects of the time and the vacuum pressure of main drying on virus titcrs had statistically significant （ P 〈 0.05 ）. The lyophilized parameters of JEV were optimized as follows ： prepare freezing temperature is at -35℃ , prepare freezing time is for 2h; main drying temperature is at -30℃→-10℃-→33℃ , main drying time is for 16h, main drying vacuum pressure is at 0. 220 mbar; final drying temperature is at 30℃, final drying vacuum pressure is at 0.001mbar, the process was not finished until the vaccum pressure is stable. The optimized freeze drying strategy of JEV has been obtained by screening the parameters of lyophilization process and it is applicable for the production after beening validated.