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J Chromatogr A. 2007 Feb 16;1142(1):2-12. Epub 2007 Jan 3.

Sorption processes in ion-exchange chromatography of viruses.

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Department of Chemical Engineering, University of Delaware, Newark, DE 19716, USA.


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.

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