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          Institute: MPI für Dynamik komplexer technischer Systeme     Collection: Bioprocess Engineering     Display Documents



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ID: 315419.0, MPI für Dynamik komplexer technischer Systeme / Bioprocess Engineering
Direct capture of influenza A virus from cell culture supernatant with Sartobind anion-exchange membrane adsorbers
Authors:Kalbfuss, B.; Wolff, M. W.; Geisler, L.; Tappe, A.; Wickramasinghe, R.; Thom, V.; Reichl, U.
Language:English
Date of Publication (YYYY-MM-DD):2007
Title of Journal:Journal of Membrane Science
Volume:299
Start Page:251
End Page:260
Review Status:Peer-review
Audience:Experts Only
Abstract / Description:Human and equine influenza A virus in cell culture supernatant (serum-free and serum-containing ultivation) was directly adsorbed to Sartobind Q and D MA75 anion-exchangers. A light scattering detector was used to trace virus particles online. The intensity of scattered light was shown to correlate with viral HA activity. Dynamic apacities of Sartobind Q (operated at a flow rate of 264 cm h-1) were consistent but depended on the cell culture medium (3.12 kHAU cm-2 for serum-free and 5.19 kHAU cm-2 for serum-containing medium). Elution of adsorbed virus from Sartobind Q by displacement with sodium chloride (up to 1.5 M, pH 7.0) resulted in average yields of 86% (based on HA activity). Elution from Sartobind D resulted in yields of only 38%. Lowering the pH (down to 4.2) or combining a low pH with salt displacement failed to elute virions. The pH of loaded supernatant was important with respect to recovery but not capacity. Preparative runs with Sartobind Q resulted in about 5-fold enrichment of HA activity and 77% reduction in total protein. Host-cell DNA, in contrast, was recovered completely in the product fraction. The average virus yield was 72%. A productivity of 67 l of culture broth m-2 h-1 was achieved. Due to their high productivity, ease of operation and acceptable yields Sartobind Q anion-exchangers can be considered promising candidates for the large-scale purification of cell culture derived influenza virus.
External Publication Status:published
Document Type:Article
Communicated by:Udo Reichl
Affiliations:MPI für Dynamik komplexer technischer Systeme/Bioprocess Engineering
External Affiliations:Sartorius AG,
Membrane Technology,
Göttingen, Germany

Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, CO, United States

Otto-von-Guericke University,
Chair of Bioprocess Engineering,
Magdeburg, Germany
Identifiers:LOCALID:21/07
URL:http://dx.doi.org/10.1016/j.memsci.2007.04.048
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