Dissolved carbon dioxide accumulation in a large scale and high density production of TGFβ receptor with baculovirus infected Sf-9 cells

From National Research Council Canada

DOIResolve DOI: https://doi.org/10.1007/BF00353924
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Name affiliation
  1. National Research Council of Canada. NRC Biotechnology Research Institute
FormatText, Article
Journal titleCytotechnology
ISSN0920-9069
1573-0778
Volume22
Issue1-3
Pages53–63
Subjectbaculovirus; bioreactor retrofitting; carbon dioxide; high density; insect cell; large scale
Abstract
Production of a TGFβ receptor with high density baculovirus infected Sf-9 cells (7×106cells ml-1) served as a test run for a retrofitted 150 L microbial fermentor. The entire 110 L batch run was performed in serum free medium, with an addition of a concentrated amino acid and yeastolate mixture at the time of infection. This addition strategy has been proven effective at a small scale by enabling cultures to maintain maximum product yield. In the bioreactor however, while cellular growth was comparable to that of the smaller scale control, TGFβ receptor production was three fold below the control. To minimize the mechanical stress, low flow rate of pure oxygen was used to control the dissolved oxygen at 40%. As a consequence, it seems that this aeration strategy involved an accumulation of dissolved carbon dioxide that in turn inhibited the protein production. A model has been developed that estimated the CO2 partial pressure in the culture to be in the vicinity of 0.15 atm. The effect of dissolved CO2 at this concentration has been assessed at smaller scale for TGFβ receptor and β-gal expression, in controlled atmosphere incubators.
Publication date
PublisherSpringer
LanguageEnglish
Peer reviewedYes
NPARC number23002044
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