Quantitation of baculovirus particles by flow cytometry

NRC publication: Ye

Analysis of baculovirus aggregates using flow cytometry

NRC publication: Ye

High yield purification of functional baculovirus vectors by size exclusion chromatography

NRC publication: Ye

Development of a simple and high-yielding fed-batch process for the production of influenza vaccines

A robust and reliable GMP-compatible fed-batch process was successfully developed for the production of recombinant hemagglutinin (rHA) proteins by expresSF\u2122 cells. The feeding solution, feeding strategy as well as the cell density at infection were optimized to maximize the final rHA production yields without affecting the existing rHA recovery protocol and downstream process. A simple and stable feeding solution was formulated and a rational feeding regimen designed to yield, depending on the rHA baculovirus used, between 2- and 3-fold enhancements in volumetric rHA production with increased specific productivity compared to the batch culture. Recombinant HA from fed-batch cultures could be simply recovered following cell lysis and purified through chromatographic steps. Overall, the increased rHA yield was maintained throughout the whole process. The performance, reproducibility and scalability of the fed-batch process was successfully demonstrated in 12 bioreactor runs of 2- and 10-L working volume using five different rHA encoding baculoviruses.Nous avons mis au point un proc\ue9d\ue9 de culture en alimentation programm\ue9e, robuste, fiable et conforme aux bonnes pratiques de fabrication (BPF) pour la production de prot\ue9ines recombinantes d\u2019h\ue9magglutinine (rHA) dans des cellules expresSF\u2122. Afin de maximiser le rendement final de production de rHA sans modifier le protocole de r\ue9colte et de traitement en aval de la prot\ue9ine, nous avons optimis\ue9 la solution et la strat\ue9gie d\u2019alimentation, ainsi que la densit\ue9 cellulaire au moment de l'infection. Nous avons \ue9labor\ue9 une solution d\u2019alimentation simple et stable ainsi qu\u2019un r\ue9gime d\u2019alimentation rigoureux permettant de doubler ou de tripler, selon le baculovirus utilis\ue9, la production volum\ue9trique tout en augmentant la productivit\ue9 sp\ue9cifique par rapport \ue0 un mode de culture discontinue. La prot\ue9ine recombinante a pu \ueatre r\ue9colt\ue9e selon un proc\ue9d\ue9 fort simple apr\ue8s la lyse des cellules cultiv\ue9es en mode semi-continu, puis purifi\ue9e par une s\ue9rie d\u2019\ue9tapes de chromatographie. Dans l\u2019ensemble, nous avons pu observer le maintien de l\u2019augmentation du rendement en rHA tout au long du proc\ue9d\ue9. Ainsi, en utilisant cinq baculovirus diff\ue9rents codant la rHA et en effectuant douze cycles de production dans des bior\ue9acteurs de 2 L et 10 L, nous avons d\ue9montr\ue9 l\u2019efficacit\ue9 du proc\ue9d\ue9 de culture semi-continue sur le plan du rendement, de la reproductibilit\ue9 et de la production \ue0 grande \ue9chelle.Peer reviewed: YesNRC publication: Ye

Monitoring motility, spreading and mortality of adherent insect cells using an impedance sensor

NRC publication: Ye

Production of Choristoneura fumiferana nucleopolyhedrovirus in C. fumiferana (CF-2C1 cells) in a 3 litre bioreactor using serum-free medium

NRC publication: Ye

Production of Recombinant Adeno-Associated Viral Vectors Using a Baculovirus/Insect Cell Suspension Cultyre System: From Shake Flasks to a 20-L Bioreactor

NRC publication: Ye