应用响应面分析法优化布氏菌活疫苗的冻干工艺参数

Optimization of freeze-drying process parameters for live Brucella vaccine using response surface methodology

目的 优化布氏菌活疫苗的冻干工艺参数。方法 采用单因素试验分析预冻温度、一次干燥持续时间和二次干燥温度对布氏菌活疫苗冻干存活率的影响;在此基础上采用响应面试验,分析3种因素不同组合试验方案的冻干存活率,拟合回归模型,探索最适合的布氏菌活疫苗冻干工艺参数。结果 优化后的布氏菌活疫苗冻干工艺参数为:预冻温度-39.9℃,时间3.75h,抽真空至0.22mbar;一次干燥温度从-20℃升温至0℃,再升至25℃,总用时16.4h,真空度0.16mbar;二次干燥温度27.3℃,时间6h,真空度0.005mbar。模型预测和实测的冻干存活率分别为74.4%和71.3%,外观、水分、浓度、活菌数等关键质量指标均符合标准。结论 响应面分析法可用于布氏菌活疫苗冻干工艺参数的优化,为后续生产过程中的冻干工艺改进提供理论参考。

Objective To optimize the freeze-drying process parameters for live Brucella vaccine.Methods We used a single parameter experiment to identify the effect of pre-freezing temperature, primary drying duration, and secondary drying temperature on the freeze-drying survival rates of live Brucella vaccine. Based on a single parameter experiment, we used a response surface experiment to determine freeze-drying survival rates for combination regimens of the three parameters, fit a regression model, and identified the most suitable freeze-drying process parameters.Results The optimized freeze-drying process parameters of live Brucella vaccine included a pre-freezing temperature-39.9℃ with a duration of 3.75 hours at a vacuum pressure of 0.22 mbar, a primary drying temperature from-20℃ to 0℃ followed by 25℃ with a total duration of 16.4 hours and a vacuum pressure of 0.16 mbar, and secondary drying temperature 27.3℃ with a duration of 6 hours and a vacuum pressure of 0.005 mbar. The freeze-drying survival rate predicted by the fitted model was 74.4%, comparable to the measured freeze-drying survival rate of 71.3%. Other key quality indicators such as appearance, moisture, concentration, and viable counts met standards.Conclusions The response surface methodology can be used to optimize freeze-drying process parameters for live Brucella vaccine. This methodology may provide a theoretical reference for improving the freeze-drying process in subsequent productions.