OP7, a novel influenza A virus defective interfering particle: production, purification, and animal experiments demonstr
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BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING
OP7, a novel influenza A virus defective interfering particle: production, purification, and animal experiments demonstrating antiviral potential Marc D. Hein 1 & Heike Kollmus 2 & Pavel Marichal-Gallardo 3 & Sebastian Püttker 1 & Dirk Benndorf 1,3 & Yvonne Genzel 3 & Klaus Schughart 2,4,5 & Sascha Y. Kupke 3 & Udo Reichl 1,3 Received: 5 September 2020 / Revised: 14 November 2020 / Accepted: 22 November 2020 # The Author(s) 2020
Abstract The novel influenza A virus (IAV) defective interfering particle “OP7” inhibits IAV replication in a co-infection and was previously suggested as a promising antiviral agent. Here, we report a batch-mode cell culture-based production process for OP7. In the present study, a seed virus containing standard virus (STV) and OP7 was used. The yield of OP7 strongly depended on the production multiplicity of infection. To inactivate infectious STV in the OP7 material, which may cause harm in a potential application, UV irradiation was used. The efficacy of OP7 in this material was preserved, as shown by an in vitro interference assay. Next, steric exclusion chromatography was used to purify and to concentrate (~ 13-fold) the UV-treated material. Finally, administration of produced OP7 material in mice did not show any toxic effects. Furthermore, all mice infected with a lethal dose of IAV survived the infection upon OP7 co-treatment. Thus, the feasibility of a production workflow for OP7 and its potential for antiviral treatment was demonstrated. Key points • OP7 efficacy strongly depended on the multiplicity of infection used for production • Purification by steric exclusion chromatography increased OP7 efficacy • OP7-treated mice were protected against a lethal infection with IAV
Keywords Influenza A virus . Antiviral . Defective interfering particles . Cell culture-based production . Steric exclusion chromatography . Animal experiments . OP7
Introduction * Sascha Y. Kupke [email protected] 1
Bioprocess Engineering, Otto von Guericke University Magdeburg, Magdeburg, Germany
2
Department of Infection Genetics, Helmholtz Centre for Infection Research, Braunschweig, Germany
3
Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Germany
4
University of Veterinary Medicine Hannover, Hannover, Germany
5
Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center, Memphis, Tennessee, USA
With annual epidemics and occasionally severe pandemics, influenza A virus (IAV, list of abbreviations, Table 1) is a major human pathogen. Every year, about 300,000–650,000 deaths are reported worldwide (Iuliano et al. 2018). The current countermeasures include the use of vaccines and antivirals like oseltamivir and zanamivir (Colman 2009; Oxford 2007; Smith et al. 2006). However, the development and production of vaccines is a time-consuming process. Moreover, resistances arising against current antivirals have been reported and are a general issue
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