Sorption processes in ion-exchange chromatography of viruses.

Sorption processes in ion-exchange chromatography of viruses.

Journal of chromatography. A (2007-01-24)

E I Trilisky, A M Lenhoff

ABSTRACT

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.

MATERIALS

Product Number

Brand

Product Description

EP951031208

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Eppendorf^® Deepwell plates, DNA LoBind, 96 wells, white plate, conical bottom, colorless wells, capacity 1000 μL, pkg of 120 ea (10 bags × 12 plates)

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Eppendorf^® Protein LoBind Microplate 384/V-PP, PCR clean, V-bottom clear wells, white border, pack of 240 ea (10 bags × 24 plates)

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Eppendorf^® DNA LoBind tubes, capacity 1.5 mL, PCR clean, pkg of 250 ea (5 x 50ea)

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Eppendorf^® Deepwell plates, DNA LoBind, 96 wells, white plate, conical bottom, colorless wells, capacity 500 μL, pkg of 40 ea (5 bags × 8 plates)

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Eppendorf^® Protein LoBind tubes, capacity 0.5 mL, PCR clean, pkg of 100 ea (2 x 50ea)

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Eppendorf^® Protein LoBind tubes, capacity 2.0 mL, PCR clean, pkg of 100 ea (2 x 50ea)

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Eppendorf^® Deepwell plates, Protein LoBind, 96 wells, yellow plate, conical bottom, colorless wells, capacity 500 μL, pkg of 40 ea (5 bags × 8 plates)

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Eppendorf^® Protein LoBind tubes, capacity 1.5 mL, PCR clean, pkg of 100 ea (2 x 50ea)

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Eppendorf^® Deepwell plates, Protein LoBind, 96 wells, white plate, conical bottom, colorless wells, capacity 500 μL, pkg of 80 ea (10 bags × 8 plates)

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Eppendorf^® Deepwell plates, Protein LoBind, 96 wells, yellow plate, conical bottom, colorless wells, capacity 500 μL, pkg of 20 ea (5 bags × 4 plates)