BIA Separations GmbH

Villach, Austria

BIA Separations GmbH

Villach, Austria
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Gerster P.,University of Natural Resources and Life Sciences, Vienna | Kopecky E.-M.,University of Natural Resources and Life Sciences, Vienna | Hammerschmidt N.,University of Natural Resources and Life Sciences, Vienna | Klausberger M.,University of Natural Resources and Life Sciences, Vienna | And 10 more authors.
Journal of Chromatography A | Year: 2013

A chromatographic process based on monoliths for purification of infective baculovirus without prior concentration step has been established. Baculovirus produced in Spodoptera frugiperda cells (Sf-9) were harvested by centrifugation, filtered through 0.8μm filters and directly loaded onto radial 1mL anion exchange monoliths with a channel size of 1.5-2.0μm operated at a volumetric flow rate of one bed volume per minute. Optional an epoxy monolith was used as pre-column to reduce interfering compounds and substances influencing the capacity of anion exchange monoliths for baculovirus infectious virus could be eluted with a step gradient at salt concentrations of 440mM NaCl. Recovery of infectious virus was highly influenced by composition and age of supernatant and ranged from 20 to >99% active baculovirus. Total protein content could be reduced to 1-8% and DNA content to 38-48% in main virus fraction. Infective virus could be 52-fold concentrated within 20.5h and simultaneously an 82-fold volume reduction was possible when loading 1150mL (2.1×108pfu/mL) onto 1mL scale support. © 2013 Elsevier B.V.

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