Sorption processes in ion-exchange chromatography of viruses
Sorption processes in ion-exchange chromatography of viruses
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DOI：
10.1016/j.chroma.2006.12.094
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发表时间：
2007-02-16
影响因子：
4.1
通讯作者：
Lenhoff, A. M.
中科院分区：
文献类型：
--
作者：
Trilisky, E. I.;Lenhoff, A. M.
纯化的病毒用于基因治疗和疫苗生产。离子交换层析（IEC）是病毒和蛋白质大规模下游纯化的最常用方法。出版的IEC协议提供了具体分离的细节，但没有选择操作参数的一般方法。为了使选择更系统，我们研究了5型腺病毒（Ad 5）作为模型病毒，并开发了批量摄取和光散射方法，用于优化吸附的离子强度和pH值，以及提供树脂几何形状。发现Ad 5的静态容量随着离子强度的增加而经历最大值。与蛋白质-树脂系统的比较表明，即使在宽孔树脂上，树脂对病毒的容量也至少低一个数量级。病毒仅部分渗透到宽孔树脂中，并且摄取速率比窄孔树脂上的摄取慢一个数量级。(c)2007 Elsevier B. V.保留所有权利。
Purified viruses are used in gene therapy and vaccine production. Ion-exchange chromatography (IEC) is the most common method for large-scale downstream purification of viruses and proteins. Published IEC protocols provide details for specific separations but not general methods for selecting operating parameters. To make the selection more systematic, we study adenovirus type 5 (Ad5) as a model virus and develop batch uptake and light scattering methods for optimizing the ionic strength and pH of adsorption, as well as providing heuristics for resin geometry. The static capacity for Ad5 was found to go through a maximum with increasing ionic strength. Comparison to a protein-resin system shows that resin capacity for the virus is at least an order of magnitude lower, even on a wide-pore resin. Virus penetration into the wide-pore resin is only partial and the uptake rate is an order of magnitude slower than the uptake onto a narrow-pore resin. (c) 2007 Elsevier B.V. All rights reserved.