Cultivation of CHO-K1 Cells for Human Enterovirus EV71 Virus-Like Particle (VLP) Production

Introduction:

Tide Motion™ CelCradle-500A (CC-500A) provides a powerful cell culture tool to achieve high cell density and high productivity of target bioproducts because of its unique feature of offering high oxygen transfer and extremely low shear stress culture environment. To evaluate this point, an interdisciplinary research collaboration was conducted in A*STAR’s Institute of Molecular and Cell Biology (IMCB) looking into the quick development of a Hand Foot and Mouth Disease (HFMD) vaccine. Through this collaboration, Esco Aster hopes to address the need for more affordable vaccines through improved biomanufacturing processes. The implementation of production scale bioreactor will deliver robust and optimized conditions for HFMD virus-like-particles (VLP) production, while providing highly consistent batch productions to IMCB.

In this study, stable CHO-K1 clones expressing Human Enterovirus (EV71) VLP was provided by Assoc. Prof. Justin Chu, IMCB, A*STAR. The application of CC-500A for the growth of CHO-K1 cells and production of Human Enterovirus (EV71) Virus-Like Particle (VLP) is elucidated. CHO-K1 cells (2×108) were seeded in CC-500A or seeded in 10 cm petri dishes (4×106). On Day 3, the cell number per CC-500A reached 1.05×109. Conditioning of Human Enterovirus (EV71) VLP with serum free media was performed on Day 3 for CC-500A or day 2 for 10 cm petri dishes. Samples were collected after three days of conditioning and Human Enterovirus (EV71) VLP amount was determined using Western Blot. CelCradle achieved a 44-fold Human Enterovirus (EV71) VLP production when compared to 2D conventional cultures with 10 cm petri dish.

This study shows CC-500A can be applied to CHO-K1 cells cultivation for Human Enterovirus (EV71) VLP production. However, the optimal condition for cultivation of CHO-K1 cells and Human Enterovirus (EV71) VLP production can be further optimized.

 

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