title: 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.
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 10l 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.
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands. Electronic address: bsander2@its.jnj.com.
authors.forename: Barbara P
authors.initials: BP
authors.lastname: Sanders
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands.
authors.forename: Isabel de los Rios
authors.initials: Ide L
authors.lastname: Oakes
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands.
authors.forename: Vladimir
authors.initials: V
authors.lastname: van Hoek
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands.
authors.forename: Wilfred
authors.initials: W
authors.lastname: Marissen
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Philip D
authors.initials: PD
authors.lastname: Minor
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Eckard
authors.initials: E
authors.lastname: Wimmer
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands.
authors.forename: Hanneke
authors.initials: H
authors.lastname: Schuitemaker
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands.
authors.forename: Jerome H H V
authors.initials: JH
authors.lastname: Custers
authors.affiliation: Division of Virology, National Institute for Biological Standards and Control, Potters Bar, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Macadam
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY, USA.
authors.forename: Jeronimo
authors.initials: J
authors.lastname: Cello
authors.affiliation: Janssen Infectious Diseases and Vaccines, Pharmaceutical Companies of Johnson and Johnson, Leiden, The Netherlands.
authors.forename: Diana
authors.initials: D
authors.lastname: Edo-Matas
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018929:Cell Culture Techniques; D002460:Cell Line; D002522:Chlorocebus aethiops; D016895:Culture Media, Serum-Free / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D011051:Poliomyelitis / Q000517:prevention & control; D017955:Poliovirus / Q000235:genetics / Q000254:growth & development*; D011054:Poliovirus Vaccine, Inactivated / Q000276:immunology; D011055:Poliovirus Vaccine, Oral; D011817:Rabbits; D014613:Vaccines, Attenuated; D014709:Vero Cells; D019562:Viral Load; D014776:Virus Cultivation / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D016895:Culture Media, Serum-Free; D011054:Poliovirus Vaccine, Inactivated; D011055:Poliovirus Vaccine, Oral; D014613:Vaccines, Attenuated
keywords: Next generation IPV; PER.C6(®) cells; Poliovirus; Rationally-attenuated strains; Sabin; Vaccines