Kitsak V Ia, Moĭsiadi S A, Bocharov A F, Landin L K, Sklianskaia E I
Vopr Virusol. 1979 Mar-Apr(2):142-8.
The results of comparative studies on concentration and purification of herpes simplex virus type 1 and type 2 (HSV-1 and HSV-2) by ficol density gradient centrifugation are presented. A two-phase distribution of extracellular HSV was established in phycol density gradient centrifugation: in zones with density of 1.110-1.114 and 1.088-1.085 g/ml. The effectiveness of purification of HSV preparations recovered from the corresponding gradient zones was determined by electron microscopy and quantitation of the contaminating cellular (radioactive) proteins in virus purification from a mixture of the culture fluid from infected cultures and the culture fluid from uninfected labeled human embryo skin-muscle tissue cultures (HESM) and a mixture of unlabeled extracellular HSV and a homogenate of labeled uninfected HESM cultures. In HSV purification from the virus-containing culture fluid the amount of cellular proteins was shown to decrease 500-fold in 150-fold virus concentration. In purification of extracellular HSV from the mixture with cell homogenate the amount of cellular proteins decreased 70- and 100-fold for HSV-2 and HSV-1, respectively. The infectious virus yield in phycol gradient centrifugation of a precipitate obtained by the addition of polyethylene glycol-6000 to the culture fluid for HSV-1 was 34.7% (in titrations in HESM cultures) and 38.4% (by intracerebral inoculation of mice weighing 5-6 g), and for HSV-2 20.2% and 26.3%, respectively.
本文展示了通过菲可密度梯度离心法对1型和2型单纯疱疹病毒（HSV-1和HSV-2）进行浓缩和纯化的比较研究结果。在菲可密度梯度离心中建立了细胞外HSV的两相分布：密度在1.110 - 1.114和1.088 - 1.085 g/ml的区域。从相应梯度区域回收的HSV制剂的纯化效果通过电子显微镜以及对从感染培养物的培养液与未感染标记的人胚胎皮肤肌肉组织培养物（HESM）的培养液混合物中纯化病毒时污染的细胞（放射性）蛋白质进行定量，以及对未标记的细胞外HSV与标记的未感染HESM培养物匀浆的混合物进行测定。在从含病毒的培养液中纯化HSV时，细胞蛋白质的量在病毒浓缩150倍的情况下减少了50倍。在从与细胞匀浆的混合物中纯化细胞外HSV时，对于HSV-2和HSV-1，细胞蛋白质的量分别减少了70倍和100倍。在通过向HSV-1培养液中添加聚乙二醇-6000获得的沉淀物的菲可梯度离心中，感染性病毒产量在HESM培养物滴定中为34.7%，在对体重5 - 6 g的小鼠进行脑内接种时为38.4%，对于HSV-2分别为20.2%和26.3%。
