Use of sulfated cellulose membrane adsorbers for chromatographic purification of cell cultured-derived influenza A and B viruses

Fortuna, Ana Raquel; Teeffelen, Sebastian Van; Ley, Adrian; Fischer, Laura M.; Taft, Florian; Genzel, Yvonne; Villain, Louis; Wolff, Michael W.; Reichl, Udo

doi:10.1016/j.seppur.2019.05.101

Cited by 22 publications

(16 citation statements)

References 36 publications

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“…This promised an independence with regard to the specific HCV genotype and the robust depletion of smaller impurities. SCMA was selected for virus polishing, based on the heparin affinity of HCV 31,34 . Additionally, both methods allowed to fully exploit the advantages of membrane adsorbers, such as a higher capacity and convective flow properties, compared to packed-bed resins 20 .…”

Section: Discussionmentioning

confidence: 99%

“…SCMA was applied for subsequent polishing, employing a pseudo affinity-based orthogonal technique. The method has been described for the purification of the Influenza A virus and the Modified Vaccinia Ankara virus [29][30][31][32][33] . It utilizes the heparin-mimicking effect of sulphated cellulose and should, thus, be widely applicable to viruses with an affinity to heparin.…”

Section: Pbsmentioning

confidence: 99%

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Development of a downstream process for the production of an inactivated whole hepatitis C virus vaccine

Lothert

Offersgaard

Pihl

et al. 2020

Sci Rep

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There is a large unmet need for a prophylactic hepatitis C virus (HCV) vaccine to control the ongoing epidemic with this deadly pathogen. Many antiviral vaccines employ whole viruses as antigens. For HCV, this approach became feasible following the development of infectious cell culture systems for virus production. However, the lack of efficient downstream processes (DSP) for HCV purification poses a roadblock for the development of a whole virus vaccine. Using cell culture-derived genotype 1a HCV we developed a scalable and efficient DSP train, employing commonly used clarification and ultrafiltration techniques, followed by two membrane-based chromatography steps. For virus capture, steric exclusion chromatography using cellulose membranes was established, resulting in a virtually complete virus recovery with > 99% protein and 84% DNA depletion. Virus polishing was achieved by sulphated cellulose membrane adsorbers with ~ 50% virus recovery and > 99% protein and 90% DNA depletion. Additional nuclease digestion resulted in 99% overall DNA depletion with final DNA concentrations of 2 ng/mL. Process results were comparable for cell culture-derived HCV of another major genotype (5a). This study provides proof-of-concept for establishment of an efficient and economically attractive DSP with potential application for production of an inactivated whole virus vaccine against HCV for human use.

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Section: Discussionmentioning

confidence: 99%

Section: Pbsmentioning

confidence: 99%

Development of a downstream process for the production of an inactivated whole hepatitis C virus vaccine

Lothert

Offersgaard

Pihl

et al. 2020

Sci Rep

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“…However, the latest studies have been performed using sulfated CEL MAs [168,171,172,178]. These pseudo-affinity MAs are made of regenerated CEL crosslinked [183] or not [182] before sulfation.…”

Section: Sulfated Cellulose Pafc Masmentioning

confidence: 99%

“…They obtained comparable results regarding the virus yield and the removal of HC-related DNA and protein impurities. MA displayed higher productivity than commercial sulfated resins [172]. Weigel et al [178] tested a two-step membrane chromatography approach to improve the purification level of IAV preparations while avoiding nuclease treatment -contrary to the two-step packed-bed chromatography process they reported previously [70] (see section 2.2.5).…”

Section: Sulfated Cellulose Pafc Masmentioning

confidence: 99%

“…The RC membrane and most Sart membranes were used in syringe filter-type units packed with 15 membrane layers (75 cm 2 total membrane area), except in the works by Peixoto et al [165] (total membrane areas: 90 cm 2 and 900 cm 2 ), Grein et al [164] (3 membrane layers, 15 cm 2 area), Nestola [177] and Weigel et al [178] where reduced membrane areas were used for small-scale experiments. For SCMA, 10 (50 cm 2 area) [43,178] or 15 (75 cm 2 area) [39] membrane layers were stacked in a stainless steel holder, while Carvalho et al [168] and Fortuna et al [171,172] used smaller membrane surfaces, i.e., 5.65 cm 2 (5 layers) [168] and 2.1 cm 2 (6 layers) [171] or 2.9 cm 2 (15 layers) [172]. c rAcMNPV, (recombinant) Autographa californica multicapsid nucleopolyhedrovirus (baculovirus); AdV-5, adenovirus type 5; rBV, (recombinant) baculovirus (derived from AcMNPV); AeDNV, Aedes aegypti densonucleosis virus; CAdV-2, canine adenovirus type 2; IAV/IBV, influenza A/influenza B virus; MVA, modified vaccinia Ankara virus.…”

Section: Conclusion and Future Trendsmentioning

confidence: 99%

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Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Junter

Lebrun

2020

Journal of Pharmaceutical Analysis

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Towards integrated production of an influenza A vaccine candidate with MDCK suspension cells

Bissinger

Marichal-Gallardo

et al. 2021

Biotech & Bioengineering

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Seasonal influenza epidemics occur both in northern and southern hemispheres every year. Despite the differences in influenza virus surface antigens and virulence of seasonal subtypes, manufacturers are well‐adapted to respond to this periodical vaccine demand. Due to decades of influenza virus research, the development of new influenza vaccines is relatively straight forward. In similarity with the ongoing coronavirus disease 2019 pandemic, vaccine manufacturing is a major bottleneck for a rapid supply of the billions of doses required worldwide. In particular, egg‐based vaccine production would be difficult to schedule and shortages of other egg‐based vaccines with high demands also have to be anticipated. Cell culture‐based production systems enable the manufacturing of large amounts of vaccines within a short time frame and expand significantly our options to respond to pandemics and emerging viral diseases. In this study, we present an integrated process for the production of inactivated influenza A virus vaccines based on a Madin–Darby Canine Kidney (MDCK) suspension cell line cultivated in a chemically defined medium. Very high titers of 3.6 log10(HAU/100 µl) were achieved using fast‐growing MDCK cells at concentrations up to 9.5 × 106 cells/ml infected with influenza A/PR/8/34 H1N1 virus in 1 L stirred tank bioreactors. A combination of membrane‐based steric‐exclusion chromatography followed by pseudo‐affinity chromatography with a sulfated cellulose membrane adsorber enabled full recovery for the virus capture step and up to 80% recovery for the virus polishing step. Purified virus particles showed a homogenous size distribution with a mean diameter of 80 nm. Based on a monovalent dose of 15 µg hemagglutinin (single‐radial immunodiffusion assay), the level of total protein and host cell DNA was 58 µg and 10 ng, respectively. Furthermore, all process steps can be fully scaled up to industrial quantities for commercial manufacturing of either seasonal or pandemic influenza virus vaccines. Fast production of up to 300 vaccine doses per liter within 4–5 days makes this process competitive not only to other cell‐based processes but to egg‐based processes as well.

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Use of sulfated cellulose membrane adsorbers for chromatographic purification of cell cultured-derived influenza A and B viruses

Cited by 22 publications

References 36 publications

Development of a downstream process for the production of an inactivated whole hepatitis C virus vaccine

Development of a downstream process for the production of an inactivated whole hepatitis C virus vaccine

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Towards integrated production of an influenza A vaccine candidate with MDCK suspension cells

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