Production of high titer attenuated poliovirus strains on the serum-free PER.C6® cell culture platform for the generation of safe and affordable next generation IPV. Issue 48 (27th November 2015)

• Record Type: Journal Article
• Authors: Sanders, Barbara P.; Oakes, Isabel de los Rios; van Hoek, Vladimir; Liu, Ying; Marissen, Wilfred; Minor, Philip D.; Wimmer, Eckard; Schuitemaker, Hanneke; Custers, Jerome H.H.V.; Macadam, Andrew; Cello, Jeronimo; Edo-Matas, Diana
• Is Part Of: Vaccine. Volume 33:Issue 48(2015)
• Page Start: 6611
• Page End: 6616
• Publication Date: 2015-11-27
• Subjects: Poliovirus -- PER.C6® cells -- Next generation IPV -- Sabin -- Rationally-attenuated strains -- Vaccines; cIPV conventional inactivated poliovirus vaccine -- DSP down stream process -- IRES internal ribosome entry site -- GPEI global poliomyelitis eradication initiative -- MOI multiplicity of infection -- OD optical density -- OPV oral poliovirus vaccine -- sIPV Sabin-based inactivated poliovirus vaccine, TCID50=tissue culture infectious dose 50% -- VAPP vaccine associated paralytic poliomyelitis; Vaccines -- Periodicals; 615.372
• DOI: 10.1016/j.vaccine.2015.10.091
• Languages: English
• ISSNs: 0264-410X

Abstract

Highlights: PER.C6® cells are susceptible to infection with attenuated poliovirus strains. Sabin strains in PER.C6® cells show increased productivity as compared to Vero cells. Sabin strain productivity in PER.C6® cells was maintained during infection at 10 l scale. Rationally-attenuated strains demonstrated high yields on the PER.C6® platform. Abstract: Background: As poliovirus eradication draws closer, alternative Inactivated Poliovirus Vaccines (IPV) are needed to overcome the risks associated with continued use of the Oral Poliovirus Vaccine and of neurovirulent strains used during manufacture of conventional (c) IPV. We have previously demonstrated the susceptibility of the PER.C6 ® cell line to cIPV strains; here we investigated the suspension cell culture platform for growth of attenuated poliovirus strains. Methods: We examined attenuated Sabin strain productivity on the PER.C6 ® cell platform compared to the conventional Vero cell platform. The suitability of the suspension cell platform for propagation of rationally-attenuated poliovirus strains (stabilized Sabin type 3 S19 derivatives and genetically attenuated and stabilized MonoCre(X) strains), was also assessed. Yields were quantified by infectious titer determination and D-antigen ELISA using either serotype-specific polyclonal rabbit sera for Sabin strains or monoclonal cIPV-strain-specific antibodies for cIPV, S19 and MonoCre(X) strains. Results: PER.C6 ® cells supported the replication of Sabin strains to yields of infectious titers that were in the range of cIPV strains at 32.5 °C. Sabin strains achieved 30-fold higher yields ( p < 0.0001) on the PER.C6 ® cell platform as compared to the Vero cell platform in infectious titer and D-antigen content. Furthermore, Sabin strain productivity on the PER.C6 ® cell platform was maintained at 10 l scale. Yields of infectious titers of S19 and MonoCre(X) strains were 0.5–1 log10 lower than seen for cIPV strains, whereas D-antigen yield and productivities in doses/ml using rationally-attenuated strains were in line with yields reported for cIPV strains. Conclusions: Sabin and rationally-attenuated polioviruses can be grown to high infectious titers and D-antigen yields. Sabin strain infection shows increased productivity on the PER.C6 ® cell platform as compared to the conventional Vero cell platform. Novel cell platforms with the potential for higher yields could contribute to increased affordability of a next generation of IPV vaccines needed for achieving and maintaining poliovirus eradication.