脊髓灰质炎病毒颗粒分离纯化方法学评价

目的比较3种不同密度梯度离心方法对脊髓灰质炎病毒空心颗粒、实心颗粒的分离效果。方法选择蔗糖密度梯度离心、氯化铯密度梯度离心、Optiprep(碘克沙醇溶液)密度梯度离心方法,对脊髓灰质炎病毒进行纯化,抽取离心之后的蛋白条带,通过计算蛋白回收率、SDS-PAGE分析、病毒颗粒透射电镜观察,评价其对病毒空心颗粒、实心颗粒的分离效果。结果病毒液经蔗糖、氯化铯和Optiprep 3种介质超速离心,均能分离空心颗粒和实心颗粒。氯化铯密度梯度离心法蛋白回收率优于Optiprep及蔗糖密度梯度离心法;Optiprep密度梯度离心法蛋白纯度优于氯化铯及蔗糖密度梯度离心法。结论通过蛋白回收率、SDS-PAGE分析、病毒颗粒透射电镜观察初步评价了3种方法对脊髓灰质炎病毒空心颗粒和实心颗粒的分离效果,为脊髓灰质炎病毒分离纯化提供了参考。

Evaluation of methods for separation and purification of poliovirus particles

Objective To compare the separation effects of empty and solid viral particles of poliovirus by sucrose density gradient centrifugation,cesium chloride density gradient centrifugation and Optiprep density gradient centrifugation.Methods Poliovirus was purified by three kinds of density gradient centrifugation methods respectively,and the protein bands were extracted. The separation effects of empty and solid viral particles were evaluated by protein recovery rate,SDS-PAGE and transmission electron microscopy. Results Both empty and solid viral particles were separated by ultracentrifugation in three media. The protein recovery rate of cesium chloride centrifugation was higher than those of Optiprep and sucrose density gradient centrifugation. However,the purity of protein purified by Optiprep centrifugation was higher than those by cesium chloride and sucrose density gradient centrifugations. Conclusion The separation effects of empty and solid viral particles by three density gradient centrifugations were preliminarily evaluated,which provided a reference for the separation and purification of poliovirus.