Search results for: Thanunthon Bowornsakulwong

Authors: Thanunthon Bowornsakulwong, Charukorn Charukarn, Franck Courtes, Panit Kitsubun, Lalintip Horcharoen

Abstract:

Vero cell is a continuous cell line that is widely used for the production of viral vaccines. However, due to its adherent characteristic, scaling up strategy in large-scale production remains complicated and thus limited. Consequently, suspension-like Vero cell culture processes based on microcarriers have been introduced and employed while also providing increased surface area per volume unit. However, harvesting Vero cells from microcarriers is a huge challenge due to difficulties in cells detaching, lower recovery yield, time-consuming and dissociation agent carry-over. To overcome these problems, we developed a dissociation-association platform technology for detaching and re-attaching cells during subculturing from microcarriers to microcarriers, which will be conveniently applied to seed trains strategies in large scale bioreactors. Herein, Hillex-2 was used to culture Vero cells in serum-containing media using spinner flasks as a scale-down model. The overall confluency of cells on microcarriers was observed using inverted microscope, and the sample cells were daily detached in order to obtain the kinetics data. The metabolites consumption and by-products formation were determined by Nova Biomedical BioprofileFlex.

Keywords: dissociation-reassociation, microcarrier, scale up, Vero cell

Procedia PDF Downloads 104

Authors: Jantakan Jullawateelert, Korakod Haonoo, Sutipong Sananseang, Sarun Torpaiboon, Thanunthon Bowornsakulwong, Lalintip Hocharoen

Abstract:

Process scale-up is essential for the biological process to increase production capacity from bench-scale bioreactors to either pilot or commercial production. Scale-up based on constant volumetric oxygen mass transfer coefficient (KLa) is mostly used as a scale-up factor since oxygen supply is one of the key limiting factors for cell growth. However, to estimate KLa of culture vessels operated with different conditions are time-consuming since it is considerably influenced by a lot of factors. To overcome the issue, this study aimed to establish correlation equations of KLa and operating parameters in 0.5 L and 5 L bioreactor employed with pitched-blade impeller and gas sparger. Temperature, gas flow rate, agitation speed, and impeller position were selected as process parameters and equations were created using response surface methodology (RSM) based on central composite design (CCD). In addition, the effects of these parameters on KLa were also investigated. Based on RSM, second-order polynomial models for 0.5 L and 5 L bioreactor were obtained with an acceptable determination coefficient (R²) as 0.9736 and 0.9190, respectively. These models were validated, and experimental values showed differences less than 10% from the predicted values. Moreover, RSM revealed that gas flow rate is the most significant parameter while temperature and agitation speed were also found to greatly affect the KLa in both bioreactors. Nevertheless, impeller position was shown to influence KLa in only 5L system. To sum up, these modeled correlations can be used to accurately predict KLa within the specified range of process parameters of two different sizes of bioreactors for further scale-up application.

Keywords: response surface methodology, scale-up, stirred-tank bioreactor, volumetric oxygen mass transfer coefficient

Procedia PDF Downloads 165