Impact of adsorbents selection on capture efficiency of cell culture derived human influenza viruses

Opitz, L.; Lehmann, S.; Zimmermann, A.; Reichl, Udo; Wolff, Michael W.

doi:10.1016/j.jbiotec.2007.07.723

Cited by 23 publications

(28 citation statements)

References 20 publications

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“…Hence, as observed for the virus particles and the dsDNA a small fraction of the loaded protein interacted with the tested matrices too strong to be desorbed during the elution process. Similar observations have been described by Opitz et al (2007a) during the purification of influenza virus particles.…”

Section: Separation Of Mva-bnsupporting

confidence: 88%

“…One possibility to economize or even to eliminate the nuclease treatment in a new production process might be the application of adsorption chromatography methods, ideally using chromatography media with predominant convective material transport like membrane adsorbers (MA). There are several examples for the application of ion exchange and affinity MA for the purification of virus particles like adenoviral vectors (Peixoto et al, 2008;Sellick, 2006), Aedes aegypti densonucleosis virus (Enden et al, 2005), baculovirus (Wu et al, 2007), and influenza virus (Kalbfuss et al, 2007a;Opitz et al, 2007aOpitz et al, , 2009. With regard to the low isoelectric point (pI) of $4.0, as estimated for the Lister strain of Vaccinia virus (Douglas et al, 1969), anion exchange MA might be a promising option.…”

Section: Introductionmentioning

confidence: 99%

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Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Wolff

Siewert

Lehmann

et al. 2009

Biotech & Bioengineering

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Smallpox is an acute, highly infectious viral disease unique to humans, and responsible for an estimated 300-500 million deaths in the 20th century. Following successful vaccination campaigns through the 19th and 20th centuries, smallpox was declared eradicated by the World Health Organization in 1980. However, the threat of using smallpox as a biological weapon prompted efforts of some governments to produce smallpox vaccines for emergency preparedness. An additional aspect for the interest in smallpox virus is its potential use as a platform technology for vector vaccines. In particular, the latter requires a high safety level for routine applications. IMVAMUNE, a third generation smallpox vaccine based on the attenuated Modified Vaccinia Ankara (MVA) virus, demonstrates superior safety compared to earlier generations and represents therefore an interesting choice as viral vector. Current downstream production processes of Vaccinia virus and MVA are mainly based on labor-intensive centrifugation and filtration methods, requiring expensive nuclease treatment in order to achieve sufficient low host-cell DNA levels for human vaccines. This study compares different ion exchange and pseudo-affinity membrane adsorbers (MA) to capture chicken embryo fibroblast cell-derived MVA-BN after cell homogenization and clarification. In parallel, the overall performance of classical bead-based resin chromatography (Cellufine sulfate and Toyopearl AF-Heparin) was investigated. The two tested pseudo-affinity MA (i.e., sulfated cellulose and heparin) were superior over the applied ion exchange MA in terms of virus yield and contaminant depletion. Furthermore, studies confirmed an expected increase in productivity resulting from the increased volume throughput of MA compared to classical bead-based column chromatography methods. Overall virus recovery was approximately 60% for both pseudo-affinity MA and the Cellufine sulfate resin. Depletion of total protein ranged between 86% and 102% for all tested matrices. Remaining dsDNA in the product fraction varied between 24% and 7% for the pseudo-affinity chromatography materials. Cellufine sulfate and the reinforced sulfated cellulose MA achieved the lowest dsDNA product contamination. Finally, by a combination of pseudo-affinity with anion exchange MA a further reduction of host-cell DNA was achieved.

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Section: Separation Of Mva-bnsupporting

confidence: 88%

Section: Introductionmentioning

confidence: 99%

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Wolff

Siewert

Lehmann

et al. 2009

Biotech & Bioengineering

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“…Hence, a projected adsorption area of about 5.6 m 2 SCM or a volume of 8.3 L CSR would be required for purification of 100 L inactivated and clarified cell culture supernatant with an HA-titer of 3. In contrast, from a membrane adsorber with the specific affinity ligand E. europaeus lectin and a dynamic binding capacity of 671 kHA/75 cm 2 , about 11 m 2 (unoptimized conditions) would be required for capturing 100 L of same cell culture supernatant (Opitz et al, 2007a (Kalbfuß et al, 2007b). This corresponds to about 389 kHAU/75 cm 2 , which is approximately threefold lower than for SCM-adsorber membranes, which was estimated from the eluted product fraction after complete saturation of the adsorber.…”

Section: Discussionmentioning

confidence: 99%

“…The fact that influenza virus envelope proteins bind to heparin was exploited using pseudo-affinity chromatography (Opitz et al, 2007a). Heparin is a heavily sulfated glycosaminoglycan consisting of hexuronic acid and D-glucosamine residues (Rabenstein, 2002).…”

Section: Introductionmentioning

confidence: 99%

“…Due to the limited pressure stability of Cellufine 1 sulfate under operating conditions (<2 bar, recommendation of manufacturer) the flow rate is restricted resulting in suboptimal productivity. Membrane adsorbers, which may overcome such drawbacks, have been already used for affinity separation of different target molecules, for example, recombinant proteins (Cattoli et al, 2006;Mellado et al, 2007), antibodies (Boi et al, 2006b;Castilho et al, 2002;Platonova et al, 1999), lectins (Boi et al, 2006a;Sorci et al, 2006) or influenza virus particles (Opitz et al, 2007a). In addition, membrane-based ion-exchange chromatography was successfully applied for purification of several other virus species, including for example insect baculoviruses (Wu et al, 2007), mosquito-specific parvoviruses (Czermak et al, 2008;Specht et al, 2004), alphaherpesviruses (Karger et al, 1998), adenovirus vectors (Peixoto et al, 2008) or influenza viruses (Kalbfuß et al, 2007b).…”

Section: Introductionmentioning

confidence: 99%

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Sulfated membrane adsorbers for economic pseudo‐affinity capture of influenza virus particles

Opitz

Lehmann

Reichl

et al. 2009

Biotech & Bioengineering

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Strategies to control outbreaks of influenza, a contagious respiratory tract disease, are focused mainly on prophylactic vaccinations in conjunction with antiviral medications. Currently, several mammalian cell culture-based influenza vaccine production processes are being established, such as the technologies introduced by Novartis Behring (Optaflu) or Baxter International Inc. (Celvapan). Downstream processing of influenza virus vaccines from cell culture supernatant can be performed by adsorbing virions onto sulfated column chromatography beads, such as Cellufine sulfate. This study focused on the development of a sulfated cellulose membrane (SCM) chromatography unit operation to capture cell culture-derived influenza viruses. The advantages of the novel method were demonstrated for the Madin Darby canine kidney (MDCK) cell-derived influenza virus A/Puerto Rico/8/34 (H1N1). Furthermore, the SCM-adsorbers were compared directly to column-based Cellufine sulfate and commercially available cation-exchange membrane adsorbers. Sulfated cellulose membrane adsorbers showed high viral product recoveries. In addition, the SCM-capture step resulted in a higher reduction of dsDNA compared to the tested cation-exchange membrane adsorbers. The productivity of the SCM-based unit operation could be significantly improved by a 30-fold increase in volumetric flow rate during adsorption compared to the bead-based capture method. The higher flow rate even further reduced the level of contaminating dsDNA by about twofold. The reproducibility and general applicability of the developed unit operation were demonstrated for two further MDCK cell-derived influenza virus strains: A/Wisconsin/67/2005 (H3N2) and B/Malaysia/2506/2004. Overall, SCM-adsorbers represent a powerful and economically favorable alternative for influenza virus capture over conventional methods using Cellufine sulfate.

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Viral Vaccines Purification

Kalbfuss

Reichl

2014

Vaccine Development and Manufacturing

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Impact of adsorbents selection on capture efficiency of cell culture derived human influenza viruses

Cited by 23 publications

References 20 publications

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Capturing of cell culture‐derived modified Vaccinia Ankara virus by ion exchange and pseudo‐affinity membrane adsorbers

Sulfated membrane adsorbers for economic pseudo‐affinity capture of influenza virus particles

Viral Vaccines Purification

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