Sokol F, Kuwert E, Wiktor T J, Hummeler K, Koprowski H
J Virol. 1968 Aug;2(8):836-49. doi: 10.1128/JVI.2.8.836-849.1968.
Extracellular rabies virus, grown in monolayer cultures of BHK21 cells in the presence of medium supplemented with bovine serum albumin, was purified by the following procedure. Virus was precipitated from infectious tissue culture fluid by zinc acetate and was resuspended in a solution of ethylenediaminetetraacetate. The suspension was filtered through a Sephadex column and was treated with ribonuclease and deoxyribonuclease. The virions were then pelleted by centrifugation at high speed and were resuspended in buffer solution. Banding of the virus by centrifugation in a sucrose density gradient was the final step in the purification procedure. Purified preparations contained bullet-shaped virus particles of variable length and little (up to 5%) contaminating host-cell material. Most of the virions were "complete", i.e., 180 nm long, but some virus particles were shorter. The length distribution of the virions was nonrandom. Shorter virions seemed to be noninfectious and showed markedly decreased hemagglutinating activity. The complement-fixing activity and the ribonucleic acid to protein ratio of the virions were not related to the length of the virus particles. Although the properties of extracellular and intracellular viruses were similar, the procedure was not suitable for purification of intracellular rabies virus.
在添加了牛血清白蛋白的培养基存在下，在BHK21细胞单层培养物中培养的细胞外狂犬病病毒，通过以下步骤进行纯化。病毒通过醋酸锌从感染性组织培养液中沉淀出来，并重悬于乙二胺四乙酸溶液中。悬浮液通过葡聚糖凝胶柱过滤，并用核糖核酸酶和脱氧核糖核酸酶处理。然后通过高速离心使病毒粒子沉淀，并将其重悬于缓冲溶液中。在蔗糖密度梯度中通过离心对病毒进行分带是纯化过程的最后一步。纯化后的制剂含有长度可变的子弹状病毒粒子，且污染的宿主细胞物质很少（高达5%）。大多数病毒粒子是“完整的”，即180纳米长，但有些病毒粒子较短。病毒粒子的长度分布不是随机的。较短的病毒粒子似乎没有传染性，并且血凝活性明显降低。病毒粒子的补体结合活性和核糖核酸与蛋白质的比率与病毒粒子的长度无关。尽管细胞外病毒和细胞内病毒的特性相似，但该方法不适用于纯化细胞内狂犬病病毒。
