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Cytotechnology

, Volume 15, Issue 1–3, pp 145–155 | Cite as

Scale-up of the adenovirus expression system for the production of recombinant protein in human 293S cells

  • Alain Garnier
  • Johanne Côté
  • Isabelle Nadeau
  • Amine Kamen
  • Bernard Massie
Article

Abstract

Human 293S cells, a cell line adapted to suspension culture, were grown to 5×106 cells/mL in batch with calcium-free DMEM. These cells, infected with new constructions of adenovirus vectors, yielded as much as 10 to 20% recombinant protein with respect to the total cellular protein content. Until recently, high specific productivity of recombinant protein was limited to low cell density infected cultures of no more than 5×105 cells/mL. In this paper, we show with a model protein, Protein Tyrosine Phosphatase 1C how high product yield can be maintained at high cell densities of 2×106 cells/mL by a medium replacement strategy. This allows the production of as much as 90 mg/L of active recombinant protein per culture volume. Analysis of key limiting/inhibiting medium components showed that glucose addition along with pH control can yield the same productivity as a medium replacement strategy at high cell density in calcium-free DMEM. Finally, the above results were reproduced in 3L bioreactor suspension culture thereby establishing the scalability of this expression system. The process we developed is used routinely with the same success for the production of various recombinant proteins and viruses.

Key words

Adenovirus human 293S cells recombinant protein scale-up metabolism 

Abbreviations

CFDMEM

calcium-free DMEM

CS

bovine calf serum

hpi

hours post-infection

J+

enriched Joklik medium

MLP

major late promoter

MOI

multiplicity of infection (# of infectious viral particle/cell)

q

specific consumption rate (mole/cell.h)

pfu

plaque forming unit (# of infectious viral particle)

Y

yield (μg/E6 cells or mole/cell)

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Copyright information

© Kluwer Academic Publishers 1994

Authors and Affiliations

  • Alain Garnier
    • 1
  • Johanne Côté
    • 1
  • Isabelle Nadeau
    • 1
  • Amine Kamen
    • 1
  • Bernard Massie
    • 1
  1. 1.Institut de recherche en biotechnologieCNRCMontréalCanada

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