应用单因素试验、复合保护剂组合试验和响应面分析法,对鳗弧菌载体疫苗MVAV6203A-1冷冻干燥的保护剂配方进行筛选和优化。试验结果表明,众多保护剂中半乳糖、海藻糖和脱脂牛奶的组合使用,能够显著提高细胞对冷冻干燥环境的耐受力。以冻干后的细胞存活率为响应值,响应面法优化的结果为:半乳糖4.6%、海藻糖2.3%、脱脂牛奶10.2%,以优化的最佳配方进行的3次重复冻干试验,细胞平均存活率可达77.3%。优化的保护剂配方适用于鳗弧菌载体疫苗的冻干生产。
The protective medium for Vibrio anguillarum vector vaccine MVAV6203A-1 was optimized by mono-factorial experiment, factor combination test and response surface methodology (RSM). The re- sults showed that the cell resistance to freeze drying was improved by combination of galactose, trehalose and skimmed milk. The quadratic model for the three significant factors was established referring cell survival rate as the target response by CCD design and response surface analysis. The optional protective me- dium was optimized as the following: 4.6% of galactose, 2.3% trehalose and 10.2% skimmed milk. In the lyophilization test repeated for 3 times, the mean V. anguillarum MVAV6203A-1 survival rate of 77.3% was achieved, indicating that the optional protective additive was suitable for the production of V. anguillarum vector vaccine MVAV6203A-1.