Lab. of Molecular Virus & Cancer, State Key Laboratory of Virology, Wuhan University School of Basic Medicine, Wuhan University, Wuhan 430071, China.
Virol J. 2010 Jun 13;7:126. doi: 10.1186/1743-422X-7-126.
Bluetongue virus (BTV) is an icosahedral non-enveloped virus within the genus Orbivirus of Reoviridae and exists as 24 distinct serotypes. BTV can infect all ruminant species and causes severe sickness in sheep. Recently, it was reported that BTV can infect some human cancer cells selectively. Because of the important oncolysis of this virus, we developed a novel purifying method for large-scale production. The purifying logic is simple, which is picking out all the components unwanted and the left is what we want. The process can be summarized in 4 steps: centrifugation, pulling down cell debrises and soluble proteins by co-immunoprecipitation with agarose Protein A, dialysis and filtration sterilization after concentration.
The result of transmission electron microscope (TEM) observation showed that the sample of purified virus has a very clear background and the virions still kept intact. The result of 50% tissue culture infective dose (TCID(50)) assay showed that the bioactivity of purified virus is relatively high.
This method can purify BTV-10 with high quality and high biological activity on large-scale production. It also can be used for purifying other BTV serotypes.
蓝舌病毒（BTV）是呼肠孤病毒科呼肠孤病毒属的二十面体无包膜病毒，存在 24 种不同的血清型。BTV 可感染所有反刍动物物种，并导致绵羊严重疾病。最近，据报道 BTV 可选择性感染某些人类癌细胞。由于该病毒具有重要的肿瘤溶解作用，我们开发了一种用于大规模生产的新型纯化方法。该纯化方法的原理很简单，即挑出所有不需要的成分，剩下的就是我们想要的。该过程可以概括为 4 个步骤：离心、用琼脂糖 Protein A 共免疫沉淀拉下细胞碎片和可溶性蛋白、透析浓缩后过滤除菌。
透射电子显微镜（TEM）观察结果表明，纯化病毒的样品背景非常清晰，病毒粒子仍保持完整。50%组织培养半数感染剂量（TCID(50)）测定结果表明，纯化病毒的生物活性相对较高。
该方法可在大规模生产中高效、高生物活性地纯化 BTV-10，也可用于纯化其他 BTV 血清型。
