冷冻干燥法制备1型糖尿病噬菌体展示疫苗  

Preparation of Phage-displayed Diabetes Type 1 Vaccine by Lyophilization

作　　者：任万军[1,2] 胡云章[1,2] 胡凝珠[1,2] 

机构地区：[1]中国医学科学院 [2]北京协和医学院医学生物学研究所疫苗研究室,昆明650118

出　　处：《中国生物制品学杂志》2010年第4期373-377,共5页Chinese Journal of Biologicals

基　　金：国家863项目"预防性I型糖尿病反义肽抗原的中试和临床前研究"(2006AA02Z4C0).

摘　　要：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 phage-displayed diabetes type 1 vaccine by lyophilization. Methods Various stabilizers were screened by orthogonal test and mixed with phage-displayed diabetes type 1 vaccine respectively, based on which the lyophilization curve was optimized. The vaccines before and after lyophilization were determined for titers and evaluated by various indexes. Results The formula of stabilizer was optimized as 10. 85% trehalose ＋ 17. 37% glycine（w / v）. The lyophilization curve was optimized as follows： S1：-40℃ 4 h, 1. 5℃ / min; S2：-15℃ 5 h, 1. 5℃ / min; S3： 25℃ 4 h, 1. 5℃ / min; vacuum： 0. 02 mbar. The decrease of vaccine titer after lyophilization was not more than 0. 5 pfu / ml. The freeze-dried vaccine showed both good appearance and good morphology under microscope, and its temperature for glass state transformation reached more than 220℃, while its residual moisture content was less than 3%. Conclusion The phage-displayed diabetes type 1 vaccine prepared by lyophilization using a stabilizer consisting of trehalose and glycine were of simple components, high thermal stability and low residual moisture content, and its lyophilization curve was simplified, while the time for storage was prolonged.

关 键 词：冷冻干燥法 1型糖尿病噬菌体展示疫苗 保护剂 

分 类 号：R587.1[医药卫生—内分泌] R392-33[医药卫生—内科学]