用加速热稳定试验筛选甲型肝炎减毒活疫苗热稳定保护剂

Selection of Thermostable Protectants of Live Attenuated Hepatitis A Vaccine with Accelerated Themostability Test

为筛选具有良好热稳定保护效果的甲肝减毒活疫苗 (H2 株 )保护剂 ,利用正交设计和加速热稳定试验 ,将不同配比的保护剂和液体疫苗混合后 ,封入安瓿 ,分别置 37℃和 45℃ ,不同时间取样 ,检测其感染性滴度 .结果表明无保护剂的对照疫苗在 45℃条件下感染性滴度仅能维持 2d不变 ,第 3天开始下降 ,至 4d已下降1.2 7logCCID50 ·mL-1;而有保护剂的 5批疫苗在 45℃条件下感染性滴度能维持 5d无显著变化 .在 37℃条件下无保护剂的对照疫苗感染性滴度仅能维持 7d无显著变化 ,10d显著变化 ,下降 1.0～ 1.1logCCID50 ·mL-1,而有保护剂的 5批疫苗在 37℃条件下感染性滴度能维持 10d无显著变化 .研究表明 0 .1mol·L-1MgSO4 ,w =5 %乳糖 ,0 .0 1mol·L-1精氨酸 ,0 .0 1mol·L-1甘氨酸组成的保护剂具有最佳保护效果 ,其热稳定保护效果明显优于现在使用的对照疫苗 .

To select the better thermostable protectants for aqueous formulation of live attenuated hepatitis A vaccine.Cross experimental design and accelerated thermostablity test were used.The control vaccine and vaccine with protectants from different combinations of lactose,MgSO 4,arginine,glycine were tested at 37℃ and 45℃.The cell culture infectious dose(CCID)were detected at different time in human diploid fibroblast(KMB17).Results showed that the control vaccine at 45℃ maintained only for 2 d without decrease of logCCID 50 ·mL -1 ,but at 4th day the control vaccine lost 1.27?logCCID 50 ·mL -1 .The vaccine with best combination of protectants showed no obvious change in logCCID 50 ·mL -1 at 45℃ for 5?d.The control vaccine at 37℃ showed no obvious decrease in logCCID 50 ·mL -1 for 1 week,but obvious decrease of 1.0—1.1?logCCID 50 ·mL -1 at 10th d.The vaccine with best combination of protectants showed no change in logCCID 50 ·mL -1 at 37℃ for 10 d.The results demonstrated that the best combination of protectants ( w =5%lactose,0.1?mol·L -1 MgSO 4,0.01?mol·L -1 arginine,0.01?mol·L -1 glycine) showed the much better thermostability than the control vaccine.