Production of Recombinant Protein Using the HeLa S3-Vaccinia Virus Expression System: Bioreactor Perfusion and Effects of Post-Infection Temperature

DOI Web Site Web Site PubMed 31 References
  • BLECKWENN Nicole A.
    Biotechnology Unit, NIDDK, National Institutes of Health Department of Chemical Engineering, University of Maryland Center for Biosystems Research, University of Maryland Biotechnology Institute
  • BENTLEY William E.
    Department of Chemical Engineering, University of Maryland Center for Biosystems Research, University of Maryland Biotechnology Institute
  • SHILOACH Joseph
    Biotechnology Unit, NIDDK, National Institutes of Health

Search this article

Abstract

Adaptation of the vaccinia virus expression system to HeLa S3 suspension bioreactor culture for the production of recombinant protein was conducted. Evaluation of hollow fiber perfusion of suspension culture demonstrated its potential for increased cell density prior to infection. The hollow fiber was also used for medium manipulations prior to infection. Two process parameters, multiplicity of infection (MOI) and temperature during the protein production phase, were evaluated to determine their effect on expression of the reporter protein, enhanced green fluorescent protein (EGFP). An MOI of 1.0 was sufficient for infection and led to the highest level of intracellular EGFP expression. Reducing the temperature to 34 °C during the protein production phase increased production of the protein two-fold compared to 37 °C in spinner flask culture. Scaling up the process to a 1.5-liter bioreactor with hollow fiber perfusion led to an overall production level of 9.9 μg EGFP/106 infected cells, or 27 mg EGFP per liter.

Journal

References(31)*help

See more

Keywords

Details 詳細情報について

Report a problem

Back to top