Simple and rapid purification of alphaherpesviruses by chromatography on a cation exchange membrane
Simple and rapid purification of alphaherpesviruses by chromatography on a cation exchange membrane
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DOI：
10.1016/s0166-0934(97)00200-0
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发表时间：
1998-02-01
影响因子：
3.1
通讯作者：
Mettenleiter, TC
中科院分区：
文献类型：
--
作者：
Karger, A;Bettin, B;Mettenleiter, TC
本文描述了一种简便、快速的纯化两种甲型疱疹病毒——伪狂犬病毒（PrV）和牛疱疹病毒1型的方法。细胞培养上清通过磺酸修饰的过滤膜，并用含氯化钾的缓冲液洗脱病毒粒子。85%以上的病毒在一个片段内被洗脱，所得病毒制剂的特异性感染性比蔗糖梯度纯化的病毒粒子高10倍以上。阳离子交换也被用于纯化一些糖蛋白缺失的PrV突变体，这些突变体在细胞培养中生长的滴度比野生型PrV低10- 100倍。对于PrV，非必需糖蛋白gC的存在，介导病毒粒子与细胞表面硫酸肝素在附着过程中的相互作用，是通过阳离子交换成功纯化的关键。(C) 1998 Elsevier Science B.V.版权所有
A simple and rapid method is described for the purification of two alphaherpesviruses, pseudorabies virus (PrV) and bovine herpesvirus 1, by chromatography on a cation exchange membrane. Cell culture supernatants were passed over a sulfonic-acid modified filter membrane and virions were eluted with a potassium chloride-containing buffer. Over 85% of the virus was eluted within a single fraction and specific infectivity of the resulting virus preparation was over 10-fold higher than that of sucrose gradient-purified virions. Cation exchange was also used for purification of PrV mutants deleted in several glycoproteins which grow in cell culture to titers 10- to 100-fold lower than those obtained by wildtype PrV. For PrV, the presence of non-essential glycoprotein gC, which mediates interaction of virions with cell surface heparin sulfate during attachment, was crucial for the successful purification by cation exchange. (C) 1998 Elsevier Science B.V. All rights reserved.