摘要
目的:对HAV病毒液的3种常见浓缩方法进行分析比较,为HAV病毒研究及规模化疫苗生产提供参考。方法:使用MILLIPORE PELLICON超滤、PEG 6000沉淀、蔗糖-甘油垫三种方法对纯化HAV病毒液进行浓缩,用ELISA方法对浓缩液进行抗原滴度检测,计算不同浓缩方法的回收率。结果:HAV病毒液经过7次超滤循环浓缩,平均回收率为86%;PEG浓缩方法回收率平均72.5%;蔗糖-甘油离心浓缩方法平均回收率53.3%。结论:蔗糖/甘油超离心法,集纯化浓缩一体,适用于样品量较少,需要高浓度样品的试验;PEG浓缩得率适中,操作简单,应用范围较广;超滤膜浓缩在大规模疫苗生产或样品量较大时适用,但需控制样品浓度及浓缩倍数不能太高,以免样品损失。
Objective： Using three common concentrated methods to enrichment hepatitis A virus, and the refined effect is analysed, concentrated compare different ways to use effect. Provide the reference for large-scale vaccine production and research of hepatitis A virus. Methods： Use MILLIPORE PELLICON ultrafiltration device, PEG6000, sucrose - glycerin cushion three methods to Hepatitis A liquid enrichment, Test concentrate antigen titer by ELISA, calculating the recovery rate of different methods. Results： Hepatitis A virus solution with ultrafiltration devices 7 times cycles, the average recovery was 86%. PEG concentration the average recovery was 72.5%; sucrose-glycerol centrifuge enrichment method average recovery was 53.3%. Conclusion： Sucrose - glycerol centrifuge enrichment method is purification concentration Synchronization and Suitable for small sample. PEG enrichment method is Simple operation, and have wide range of applications. Ultrafiltration device can be effectively concentrated for large scale production, but when the sample titer is high, it lead to sample loss.
出处
《现代生物医学进展》
CAS
2014年第24期4616-4619,共4页
Progress in Modern Biomedicine
基金
云南省应用基础研究面上项目(2010ZC230)
关键词
HAV
超滤浓缩
回收率
Hepatitis A virus
Ultrafiltration concentrated
Recovery rate
