使用高效尺寸排阻色谱法分离脊髓灰质炎病毒和亚病毒颗粒。

Use of high-performance size-exclusion chromatography for the separation of poliovirus and subviral particles.

作者信息

Foriers A, Rombaut B, Boeyé A

机构信息

Department of Medical and Special Biochemistry, Vrije Universiteit Brussel, Belgium.

出版信息

J Chromatogr. 1990 Jan 5;498(1):105-11. doi: 10.1016/s0021-9673(01)84239-4.

DOI:10.1016/s0021-9673(01)84239-4

PMID:2153149

Abstract

The size-dependent separation of viral and subviral particles in the range 10(5)-10(7) daltons was undertaken by high-performance liquid chromatography. A combination of Ultrahydrogel 2000 and 1000 size-exclusion columns, equilibrated and developed with Tris buffer (pH 7.4), was used to fractionate extracts of cells infected with radiolabelled poliovirus. Poliovirions (30 nm) and subviral particles (20 nm) were separated according to size with full retention to their biological activities. Procapsids (same size as virions, but devoid of RNA) could not be separated from virions. Sample recoveries as determined with radiolabelled material constantly exceeded 70%. The method was successfully applied to the separation of viral and subviral particles from complex mixtures.

摘要

采用高效液相色谱法对分子量在10⁵ - 10⁷道尔顿范围内的病毒和亚病毒颗粒进行了尺寸依赖性分离。使用Ultrahydrogel 2000和1000尺寸排阻柱的组合，用Tris缓冲液（pH 7.4）平衡并展开，对感染了放射性标记脊髓灰质炎病毒的细胞提取物进行分级分离。脊髓灰质炎病毒颗粒（30纳米）和亚病毒颗粒（20纳米）根据尺寸进行分离，并完全保留其生物活性。前衣壳（与病毒颗粒大小相同，但不含RNA）无法与病毒颗粒分离。用放射性标记物质测定的样品回收率始终超过70%。该方法成功应用于从复杂混合物中分离病毒和亚病毒颗粒。