Steric exclusion chromatography for purification of cell culture-derived influenza A virus using regenerated cellulose membranes and polyethylene glycol

Marichal-Gallardo, Pavel; Pieler, Michael; Wolff, Michael W.; Reichl, Udo

doi:10.1016/j.chroma.2016.12.076

Cited by 61 publications

(85 citation statements)

References 59 publications

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“…The infectious virus titer was quantified with a TCID 50 assay according to Genzel and Reichl (2007). Host cell dsDNA and total protein concentrations were measured with a Picogreen and a Bradford assay, respectively (Marichal-Gallardo et al 2017). The distribution of largesized virus and other aggregates was measured using a disc centrifuge (CPS DC24000 UHR disc centrifuge, CPS Instruments Inc., LA, USA) following a method described earlier ).…”

Section: Analyticsmentioning

confidence: 99%

Performance of an acoustic settler versus a hollow fiber–based ATF technology for influenza virus production in perfusion

Gränicher

Coronel

Trampler³

et al. 2020

Appl Microbiol Biotechnol

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Process intensification and integration is crucial regarding an ever increasing pressure on manufacturing costs and capacities in biologics manufacturing. For virus production in perfusion mode, membrane-based alternating tangential flow filtration (ATF) and acoustic settler are the commonly described cell retention technologies. While acoustic settlers allow for continuous influenza virus harvesting, the use of commercially available membranes for ATF systems typically results in the accumulation of virus particles in the bioreactor vessel. Accordingly, with one single harvest at the end of a cultivation, this increases the risk of lowering the product quality. To assess which cell retention device would be most suitable for influenza A virus production, we compared various key performance figures using AGE1.CR.pIX cells at concentrations between 25 and 50 × 10 6 cells/mL at similar infection conditions using either an ATF system or an acoustic settler. Production yields, process-related impurities, and aggregation of viruses and other large molecules were evaluated. Taking into account the total number of virions from both the bioreactor and the harvest vessel, a 1.5-3.0-fold higher volumetric virus yield was obtained for the acoustic settler. In addition, fewer large-sized aggregates (virus particles and other molecules) were observed in the harvest taken directly from the bioreactor. In contrast, similar levels of process-related impurities (host cell dsDNA, total protein) were obtained in the harvest for both retention systems. Overall, a clear advantage was observed for continuous virus harvesting after the acoustic settler operation mode was optimized. This development may also allow direct integration of subsequent downstream processing steps. Key points• High suspension cell density, immortalized avian cell line, influenza vaccine.

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Section: Analyticsmentioning

confidence: 99%

Performance of an acoustic settler versus a hollow fiber–based ATF technology for influenza virus production in perfusion

Gränicher

Coronel

Trampler³

et al. 2020

Appl Microbiol Biotechnol

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“…Besides continuous chromatography, steric exclusion chromatography (SXC) has also been proposed for purification of Influenza virus and Bacteriophages [141,142]. The principle of this technique is the capturing of the target particles based on the mutual steric exclusion of polyethylene glycol (PEG) between the product and a non-reactive hydrophilic surface.…”

Section: Innovations In Purification Technologiesmentioning

confidence: 99%

“…In SXC, there is the mixing of the feedstock material, previously clarified, with polyethylene glycol (PEG) at a defined concentration and molecular weight (MW), and the respective loading into the non-reactive hydrophilic matrix. Influenza viruses were purified with SXC with recoveries above 95% and depletion of host cell DNA and total protein above 99% and 92%, respectively [142].…”

Section: Innovations In Purification Technologiesmentioning

confidence: 99%

Current challenges in biotherapeutic particles manufacturing

Moleirinho

Silva

Alves

et al. 2019

Expert Opinion on Biological Therapy

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Introduction: The development of novel complex biotherapeutics led to new challenges in biopharmaceutical industry. The potential of these particles has been demonstrated by the approval of several products, in the different fields of gene therapy, oncolytic therapy, and tumor vaccines. However, their manufacturing still presents challenges related to the high dosages and purity required. Areas covered: The main challenges that biopharmaceutical industry faces today and the most recent developments in the manufacturing of different biotherapeutic particles are reported here. Several unit operations and downstream trains to purify virus, virus-like particles and extracellular vesicles are described. Innovations on the different purification steps are also highlighted with an eye on the implementation of continuous and integrated processes. Expert opinion: Manufacturing platforms that consist of a low number of unit operations, with higheryielding processes and reduced costs will be highly appreciated by the industry. The pipeline of complex therapeutic particles is expanding and there is a clear need for advanced tools and manufacturing capacity. The use of single-use technologies, as well as continuous integrated operations, are gaining ground in the biopharmaceutical industry and should be supported by more accurate and faster analytical methods.

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“…Lee et al [186] illustrated the separation efficiency of this technique by purifying immunoglobulin G (IgG) and bacteriophage M13K07, using hydroxyl-substituted CIM® polymethacrylate monoliths as a hydrophilic surface. SEX has been implemented recently in the purification of cell culture-derived IAV using simple regenerated CEL MA (1 µm pore size membrane filters) as an alternative to monoliths [173]. High virus recoveries (>95%) were reached.…”

Section: Cellulose Steric Exclusion Chromatography Mamentioning

confidence: 99%

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

Junter

Lebrun

2020

Journal of Pharmaceutical Analysis

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Steric exclusion chromatography for purification of cell culture-derived influenza A virus using regenerated cellulose membranes and polyethylene glycol

Cited by 61 publications

References 59 publications

Performance of an acoustic settler versus a hollow fiber–based ATF technology for influenza virus production in perfusion

Performance of an acoustic settler versus a hollow fiber–based ATF technology for influenza virus production in perfusion

Current challenges in biotherapeutic particles manufacturing

Polysaccharide-based chromatographic adsorbents for virus purification and viral clearance

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