优化使用膜色谱法浓缩和纯化逆转录病毒。
Optimised concentration and purification of retroviruses using membrane chromatography.
机构信息
Department of Chemical Engineering and Biotechnology, New Museums Site, Pembroke St, Cambridge CB2 3RA, UK.
King's College London, 123 Coldharbour Lane, London SE5 9NU, UK.
出版信息
J Chromatogr A. 2014 May 2;1340(100):24-32. doi: 10.1016/j.chroma.2014.03.023. Epub 2014 Mar 15.
The ability of an anion exchange membrane to purify a γ-retrovirus was assessed and optimised with respect to different loading and wash buffers. Recoveries of infectious virus greater than 50% were consistently obtained, while specific titre was increased up to one thousand fold when compared to the material loaded. Specific proteins removed and retained by this optimised process were identified by mass spectrometry. It was possible to successfully bind and elute the equivalent of 1.27 × 10(8) Ifu/ml of ion exchange membrane. This could then be highly concentrated, with infectious virus concentrated to a maximum of 420-fold compared to the load.
评估了阴离子交换膜对γ逆转录病毒的净化能力，并针对不同的加载和洗涤缓冲液对其进行了优化。与加载的材料相比，始终获得大于 50%的感染性病毒回收率，而特定效价增加了一千倍。通过质谱鉴定了通过此优化过程去除和保留的特定蛋白质。成功地结合和洗脱了相当于 1.27×10(8)Ifu/ml 的离子交换膜。然后可以将其高度浓缩，与负载相比，感染性病毒浓缩最多可达 420 倍。
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