一种用于流感病毒亚单位疫苗纯度分析的改进方法

摘要

目的：在SDS-PAGE方法的基础上建立一种改进方法,用于流感病毒亚单位疫苗中间体的杂质分析。方法：用糖苷酶F对不同亚型的疫苗中间体去糖基化处理后进行SDS-PAGE,与未经去糖基化处理的还原型SDS-PAGE结果进行比较。结果：去糖基化处理消除了血凝素的糖基化对电泳迁移率的影响,从而实现了血凝素亚基和杂质的电泳分离。结论：此方法应用于流感病毒亚单位疫苗中间体的杂质分析,较之常规的还原型SDS-PAGE方法具有更高的分辨力。%Objective: To establish an improved method of impurity analysis based on SDS-PAGE for the inter⁃mediate of influenza subunit vaccine. Methods: Different types of vaccine intermediates were deglycosylated using PNGase F followed by SDS-PAGE. Deglycosylated and non-deglycosylated samples were analyzed by SDS-PAGE. Results: Deglycosylating eliminate the effects of electrophoretic mobility by haemagglutinin glycosylation. Therefore, subunits of haemagglutinin could be separated from impurities satisfactorily by electrophoresis. Conclusion: This improved method provided a higher resolution in the impurity analysis for influenza subunit vaccine intermediate, in comparison of regular reduced SDS-PAGE.