One of the challenges in the vaccine industry is to produce large quantities of vaccines in a rapid and cost-effective way. Major changes to current bioprocesses are difficult to implement due to costs associated with the regulatory approval process. Therefore, we propose to use insulin, a growth factor currently used and approved in several processes and production platforms, as an additive to boost cell growth, productivity and viral yield.
A HEK293 clone, HEK 293SF-3F6 has been adapted to growth in suspension in serum-free conditions. In a commercially available media, the cell density was increased by 4-fold upon addition of 10mg/L insulin. In a media formulated in-house, the growth was accelerated with insulin and we reached maximal cell density within 6 days instead of 10 days.
HEK293SF-3F6 cells constitute a permissive host for influenza viruses and several different strains have been produced. Adenoviruses (AdV) have also successfully been produced in large-scale bioreactors using HEK293SF-3F6 cells. In order to improve production of viruses and viral vectors, the addition of increasing concentrations of insulin at different time points during the infection process has been examined. Traditional quantification methods used to titer influenza are variable and low-throughput.
In this webinar, we will also describe an approach that allows rapid and robust quantification of multiple strains of influenza using universal antibodies in conjunction with a dot blot technique. When 25 to 100µg/ml insulin was added to the culture at the time of infection, the productivity of an H1N1 influenza strain was increased by nearly two-fold. This increase in production was accompanied by an activation of signaling pathways associated with cell survival. Results with an H3N2 strain will also be presented. Finally, the effect of insulin on AdV production will be discussed and an example of improved production in a 3L-bioreactor with insulin will be shown.
Aziza Manceur,
Research Officer
Aziza is a research officer at the National Research Council Canada, a Research and Technology organization funded by the government of Canada. She is part of the Cell Culture Scale Up team, a team that specializes in the large production of viral vectors and proteins using mammalian cells. She earned her PhD in biomedical engineering from the University of Toronto and went to the University of Pennsylvania for a post-doctorate training.
- Learn about the effects of insulin on cell growth and virus production
- Description of a robust method for influenza quantification with universal antibodies, and adenovirus using HPLC
- Gain increased understanding of large-scale production of viral vectors using HEK293SF-3F6 cells in bioreactors
- Research & Development
- Process Development
- Researchers
- Scientists
- Principal Investigator
- Lab Managers
