Adenovirus Vector Production in 293 Fed-Batch Cultyres

Wong, Kainam Thomas; Guardia, María Jesús; Lee, Stanford; Hu, Wei‐Shou

doi:10.1007/0-306-46875-1_11

Cited by 2 publications

(2 citation statements)

References 3 publications

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“…The control of glucose and glutamine at low levels has allowed for an increase in the density of infection and has led to higher virus titers (Wong et al, 1999). However, a reduction in specific production was noted, hypothesized to result from a decrease in infection efficiency due to cell aggregate formation.…”

Section: Introductionmentioning

confidence: 98%

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

Henry

Dormond

Perrier

et al. 2004

Biotech & Bioengineering

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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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Section: Introductionmentioning

confidence: 98%

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

Henry

Dormond

Perrier

et al. 2004

Biotech & Bioengineering

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“…The development of cell culture processes to increase adenovirus production has been often approached by designing feeding strategies that improve nutrient supply and reduce the accumulation of inhibitory metabolite(s) [7,24]. Since most commercial culture media are based on proprietary formulations, very few data have been published on nutritional requirements during cell growth as well as virus production.…”

Section: Discussionmentioning

confidence: 99%

Optimization of Culture Media and Feeding Strategy for High Titer Production of an Adenoviral Vector in HEK 293 Fed-Batch Culture

Shen,

Rodenbrock,

Lanthier

et al. 2024

Vaccines

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Adenoviruses are efficient and safe vectors for delivering target antigens and adenovirus-based vaccines have been used against a wide variety of pathogens, including tuberculosis and COVID-19. Cost-effective and scalable biomanufacturing processes are critical for the commercialization of adenovirus-vectored vaccines. Adenoviral vectors are commonly produced through the infection of batch cultures at low cell density cultures, mostly because infections at high cell densities result in reduced cell-specific virus productivity and does not improve volumetric productivity. In this study, we have investigated the feasibility of improving the volumetric productivity by infecting fed-batch cultures at high cell densities. Four commercial and one in-house developed serum-free media were first tested for supporting growth of HEK 293 cells and production of adenovirus type 5 (Ad5) in batch culture. Two best media were then selected for development of fed-batch culture to improve cell growth and virus productivity. A maximum viable cell density up to 16 × 106 cells/mL was achieved in shake flask fed-batch cultures using the selected media and commercial or in-house developed feeds. The volumetric virus productivity was improved by up to six folds, reaching 3.0 × 1010 total viral particles/mL in the fed-batch culture cultivated with the media and feeds developed in house and infected at a cell density of 5 × 106 cells/mL. Additional rounds of optimization of media and feed were required to maintain the improved titer when the fed-batch culture was scaled up in a bench scale (3 L) bioreactor. Overall, the results suggested that fed-batch culture is a simple and feasible process to significantly improve the volumetric productivity of Ad5 through optimization and balance of nutrients in culture media and feeds.

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Adenovirus Vector Production in 293 Fed-Batch Cultyres

Cited by 2 publications

References 3 publications

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

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

Optimization of Culture Media and Feeding Strategy for High Titer Production of an Adenoviral Vector in HEK 293 Fed-Batch Culture

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