Metabolic Profiling of Chinese Hamster Ovary Cell Cultures at Different Working Volumes and Agitation Speeds Using Spin Tube Reactors
Dr Mark Elvin discusses his recent publication:
In this paper, just published in late 2020, myself and colleagues from The Manchester Institute of Biotechnology investigated the effect of varying the starting cell culture volume and agitation speed and the impact this had on cell growth, recombinant protein production and cellular metabolism of two, clonally-derived Chinese Hamster Ovary (CHO) cell lines. This study provides a better understanding of the consequences of the starting cell culture working volume and agitation speed on CHO cell culture performance and metabolism. The findings show that cell culture parameters such as culture volume and agitation speed have a significant impact on the metabolism and culture performance of recombinant CHO cell lines. Key findings from the study were:
- By altering the starting working culture volume, cells rebalanced cell growth and recombinant protein production. Low culture volumes increased recombinant protein production, but decreased cell growth, while high culture volumes had the opposite effect.
- By adjusting the agitation speeds of the cultures, cells either exacerbated or moderated the differences observed due to the culture volume changes. Low agitation speeds suppressed cell growth and recombinant protein production in high volume cultures, whereas high agitation speeds closed the differences in culture performance between low and high culture volumes.
- These differences were also reflected in their cellular metabolism as low culture volumes enhanced oxidative metabolism (linked to a productive phenotype) whilst high culture volumes generated a metabolic profile that was more predominately glycolytic (linked to a more proliferative phenotype).
These findings indicate that the culture volume influence on metabolism modulates the balance between cell growth and recombinant protein production, a key feature that maybe useful to adjust CHO cells, and potentially other cell types, toward a more productive phenotype.
As a CRO, Peak Proteins offers clients the choice of mammalian, insect or E.coli-based expression systems in order to produce their proteins – the choice of expression system is dependent on the type of protein to be expressed (e.g. secreted, intracellular, correct type of post-translational modifications etc.). It will be interesting to follow up on this study at Peak Proteins, using the understanding and knowledge gained and apply it to our own in-house expression systems, especially our HEK293 suspension system. This cell line is the most popular choice when it comes to transient mammalian expression due to increased transfection efficiency over other transient mammalian expression systems.
