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Phenotyping of mammalian CHO cell line in fed-batch bioprocess by intensified and classical procedure : master's thesis
Matic Košir, 2023, master's thesis

Abstract: Nowadays, pharmaceutical industry is striving to find more biologically relevant solutions and to move from synthetic to biological products. Biopharmaceuticals represent an emerging sector in the pharmaceutical industry and are useful for a wide range of indications, including oncology and rheumatology. The development and optimisation of bioprocesses at laboratory scale is crucial for process understanding and translation into production. Bioprocess development often begins with comparison of several potential clones to be characterized, tested in low-volume cultures with different media or other variables, and then evaluated for growth, productivity, and consistency. The purpose of this master's thesis is to review changes in growth curves, substrate consumption, yield, and product quality of a mammalian cell line when substituting glucose as the main carbon source with alternative energy sources. This master thesis presents an attempt to adapt a mammalian CHO cell line to alternative energy sources. A comparison of the cultivation with maintenance of the CHO mammalian cell line by an intensified and a classical process is also shown. Substrate requirements at different time points of the process were monitored using phenotypic microarrays. Based on the obtained and analyzed results of the phenotypic microarrays, we tried to optimize the cell culture supplementation strategy. We have tried to use small molecules to facilitate adaptation to alternative energy sources. Our results showed that growing CHO cell culture on an alternative energy source-maltose and cell reprogramming with small molecules affects the quality of the product. Trends in cell growth, substrate consumption, by-product excretion and final product concentration are similar between the reference cell cultures and those pre-treated with small molecules. We found that adapting a mammalian CHO cell line to an alternative energy source is a challenging process that would need to be carried out over a long period of time.
Keywords: CHO cells, bioprocess, phenotyping, alternative energy sources, glycan mapping
Published in DKUM: 05.10.2023; Views: 182; Downloads: 73
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Determination of the process parameters relative influence on k[sub]La value using Taguchi design methodology
Marko Tramšek, Andreja Goršek, 2007, original scientific article

Abstract: This article describes experimental determination of the relative impact of significant process parameters that influence volumetric oxygen mass transfer coefficient (kLa) using Taguchi design methodology. For this purpose an automated RC1 reaction calorimeter (Mettler-Toledo), which was originally developed for chemical processes, was modified for the bioprocesses. Simple fermentation using Baker's yeast was studied to illustrate the design procedure. Orthogonal array L25 was selected for the proposed design and ANOVA method was used for recognizing the relative influence of the process parameters. Within the observed range of temperature (?), fermentation media volume (VFM), and yeast mass concentration (?Y), these process parameters were found to be unimportant compared to the volumetric air flow rate (qV,a) and rotational frequency of the stirrer (fm). The qV,a had a substantial effect on the kLa value (89.2 %) and the fm had just a small one (3.6 %), meanwhile the remain fraction to 100 % represents error. The results refer strictly to the selected case study. Anyhow, the proposed procedure shows that application of the Taguchi approach for analyzing the oxygen mass transfer based on the experimental data obtained from a highly-automated laboratory reactor appears to have potential usage in general biopharmaceutical process design.
Keywords: bioprocess parameters, mass transfer, volumetric oxygen mass transfer coefficient, process parameters, Taguchi method, analysis of variance
Published in DKUM: 31.05.2012; Views: 2378; Downloads: 51
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