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Cytotechnology

, Volume 49, Issue 2–3, pp 161–171 | Cite as

Development of a Serum-free Suspension Process for the Production of a Conditionally Replicating Adenovirus using A549 Cells

  • Robert LongleyJr.
  • Laurie Radzniak
  • Marc Santoro
  • Yung-Shyeng Tsao
  • Russell G. G. Condon
  • Peggy Lio
  • Marcio Voloch
  • Zhong LiuEmail author
Research article

Abstract

Conditionally replicating adenoviruses (CRAVs) are a group of recombinant human adenoviruses genetically engineered to replicate in selected tissues, such as tumors. These viruses could potentially offer significant medicinal benefits, since the restrictive replication of these viral vectors leads to the lysis of target cells without harm to the surrounding tissues. The in vitro propagation and amplification of the CRAV vectors often requires special host cells with deregulated growth control pathways. In order to develop an efficient cell culture process for the scaleable production of a CRAV vector, A549 cells, a human lung carcinoma cell line normally cultured in adherent culture, were adapted to suspension culture. CRAV production was demonstrated with the suspension-adapted A549 cells and a baseline production process was developed in shake flasks. The ability to scale-up virus production was confirmed in stirred tank bioreactors. Molecular characterization of the suspension-adapted A549 cells indicates no significant changes in cellular mechanisms related to adenovirus infection.

Keywords

Adenovirus Recombinant adenovirus Gene therapy A549 cells Serum-free Suspension Conditionally replicating adenoviruses (CRAVs) 

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Copyright information

© Springer 2006

Authors and Affiliations

  • Robert LongleyJr.
    • 1
  • Laurie Radzniak
    • 1
  • Marc Santoro
    • 1
  • Yung-Shyeng Tsao
    • 1
  • Russell G. G. Condon
    • 1
  • Peggy Lio
    • 1
  • Marcio Voloch
    • 1
  • Zhong Liu
    • 1
    Email author
  1. 1.Biotechnology DevelopmentSchering-Plough Research InstituteUSA

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