论文部分内容阅读
目的采用冷冻干燥法制备1型糖尿病噬菌体展示疫苗。方法将正交试验筛选的不同冻干保护剂与1型糖尿病噬菌体展示疫苗混合,优化冻干曲线进行冻干。检测冻干前后噬菌体疫苗的滴度,并通过各项指标对冻干后的疫苗进行评价。结果经筛选,最佳保护剂配方为:10.85%海藻糖+17.37%甘氨酸(w/v);最佳冻干曲线为:S1:-40℃4h,1.5℃/min;S2:-15℃5h,1.5℃/min;S3:25℃4h,1.5℃/min;真空度:0.02mbar。冻干后疫苗滴度下降不超过0.5pfu/ml,外观及电镜观察疫苗样品形态均较好,玻璃态转化温度能达到220℃以上,含水量小于3%。结论以海藻糖、甘氨酸为保护剂,采用冷冻干燥法制备的1型糖尿病噬菌体展示疫苗具有保护剂组成成分少、热稳定性强、含水量低、冻干曲线简化、保存时间长等特点。
Objective To prepare a type 1 diabetes phage display vaccine by freeze-drying. Methods The different freeze-dried protective agents selected by orthogonal test were mixed with the type 1 diabetes phage display vaccine, and the lyophilization curve was optimized for lyophilization. The phage vaccine titer was tested before and after lyophilization, and the freeze-dried vaccine was evaluated by various indexes. Results: The best protective agent formula was: 10.85% trehalose and 17.37% glycine (w / v); the best lyophilization curve was: S1: -40 ℃ 4h, 1.5 ℃ / min; S2: -15 ℃ 5h , 1.5 ° C / min; S3: 4h at 25 ° C, 1.5 ° C / min; degree of vacuum: 0.02 mbar. After the freeze-dried, the titer of the vaccine did not drop more than 0.5pfu / ml. The appearance and electron microscopy were all good. The glass transition temperature was above 220 ℃ and the water content was less than 3%. Conclusion The type 1 diabetes phage display vaccine prepared by lyophilization with trehalose and glycine as protective agent has the characteristics of less components of protective agent, high thermal stability, low water content, simplified lyophilization profile and long storage time.