Insights into adenoviral vector production kinetics in acoustic filter-based perfusion cultures.

Biotechnol Bioeng

Ecole Polytechnique de Montréal, Montréal, Québec, Canada.

Published: June 2004

AI Article Synopsis

  • The production of adenoviral vectors faces challenges due to decreased productivity at higher cell densities, likely due to nutrient depletion or metabolite inhibition.
  • To address this, researchers explored perfusion as a method to enhance productivity by allowing for effective infection in HEK-293 cell cultures at higher densities, using an acoustic cell separator to minimize cellular damage.
  • The study found that perfusion maintained similar cell-specific productivity compared to lower-density batch cultures and achieved significantly higher virus concentrations, with real-time monitoring available to assess infection status and nutrient dynamics.

Article Abstract

One of the major limitations in the production of adenoviral vectors is the reduction in cell-specific productivity observed for increasing cell density at infection in batch cultures. This observation strongly suggests some nutrient depletion and/or metabolite inhibition in the media. These limitations have been partially overcome through other feeding strategies, such as fed-batch and sequential batch operations. To improve these results, we evaluated perfusion as a strategy to increase the volumetric productivity of HEK-293 cell cultures, by allowing productive infection at higher cell densities. An acoustic cell separator was employed in consideration of the increased shear sensitivity of the cells during the infection phase. The effects of perfusion rate and cell density at infection on the production of a recombinant adenovirus expressing the GFP were investigated. The perfusion mode allowed successful infection at cell densities in the range of 2.4-3 x 10(6) cell/mL, while maintaining a similar cell specific productivity (17,900 +/- 2400 VP/cell) to that of a batch infected at a low cell density (5 x 10(5) cell/mL). The highest virus concentrations (4.1 +/- 0.6 x 10(10) VP/mL) were attained for a feed rate of 2 vol/d and constituted a fivefold increase compared to a batch with medium replacement. Rapid assessment of the infection status was achieved through the use of on-line monitoring of respiration, fluorescence, and biovolume. Analysis of the kinetics of nutrient consumption and metabolite production revealed that a reduction in specific productivity is correlated with reduced metabolic activity.

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Source
http://dx.doi.org/10.1002/bit.20074DOI Listing

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