- Poster presentation
- Open Access
Benchmarking of commercially available CHO cell culture media for antibody production
- David Reinhart1Email author,
- Christian Kaisermayer2,
- Lukas Damjanovic1 and
- Renate Kunert1
https://doi.org/10.1186/1753-6561-7-S6-P13
© Reinhart et al.; licensee BioMed Central Ltd. 2013
- Published: 4 December 2013
Keywords
- Cell Concentration
- Antibody Production
- Chinese Hamster Ovary Cell
- Chinese Hamster Ovary
- Product Concentration
Introduction
Chinese hamster ovary (CHO) cells have become the preferred expression system for the production of complex recombinant proteins. Several suppliers offer CHO specific cell cultivation media and sometimes also media systems, which combine feeds and basal medium. We compared eight commercially available CHO cell culture media and feed supplements from three different vendors to evaluate their influence on cell growth and antibody production of a CHO cell line. In conclusion, ActiCHO™ Media System, with a matching base media and feeds, resulted in the highest cell growth and the highest productivity. Further nutrient additions did not have a profound effect on the process performance.
Materials and methods
Cultivation media:
ActiCHO P (GE Healthcare)
CD CHO (Life Technologies)
CD OptiCHO™ (Life Technologies)
CD FortiCHO™ (Life Technologies)
Ex-Cell™ CD CHO (Sigma Aldrich)
ProCHO 5 (Lonza)
BalanCD™ CHO Growth A (Irvine Scientific)
Cellvento™ CHO-100 (EMD Millipore)
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Anti-Clumping Agent (Life Technologies)
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CHO DG44 cells expressing an IgG antibody
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Cultivation conditions: 37°C, 7% CO2, 140 rpm
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Batch and fed-batch cultivations were run in Erlenmeyer shake flasks (Corning, NY). The cultures were grown in a CO2 incubator shaker (Kühner, Switzerland)
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Batch cultures were run as single experiments, the method variability was determined by a triplicate reference experiment in ActiCHO P.
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During fed-batch processes the cultures were fed with the corresponding feeds ActiCHO Feed A and Feed B (GE Healthcare), BalanCD™ CHO Feed 1 (Irvine Scientific) or EfficientFeed™ A and/or FunctionMAX™ (both Life Technologies) according to the manufacturers inctructions [1]. The respective feeding regimens are shown in Table 1.
Feeding regimens in fed-batch cultures.
Basal medium | ActiCHO Feed A | ActiCHO Feed B | EfficientFeed A | FunctionMAX | Feed 1 | Peak cell conc. [106 c/ml] | Harvest Titer [g/L] |
---|---|---|---|---|---|---|---|
ActiCHO P | daily; 3% | daily; 0.3% | - | - | 23.9 | 5.48 | |
ActiCHO P | daily; 3% | daily; 0.3% | - | 3, 5, 7; 3.3% | 21.3 | 5.82 | |
CD OptiCHO | - | - | 3, 5, 7, 9; 10% | - | 5.8 | 0.72 | |
CD OptiCHO | - | - | 3, 5, 7; 10% | - | 5.2 | 0.80 | |
CD OptiCHO | - | - | 3, 5, 7; 10% | 3, 5, 7; 3.3% | 6.3 | 1.74 | |
CD OptiCHO | daily; 3% | daily; 0.3% | - | - | 9.0 | 1.46 | |
BalanCD CHO | - | - | - | - | 1, 3, 5; 10% | 7.1 | 1.30 |
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Fed-batch cultures were run in triplicates. The residual glucose concentration was maintained above 3 g/L by addition of glucose concentrate
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Analytics: cell concentration, viability, selected metabolites, product concentration, amino acid concentrations
Results and discussion
Cell concentrations (upper panel) and product concentrations (lower panel) obtained in batch experiments with different commercially available CHO cell culture media. Titers in CD FortiCHO were not determined due to low cell concentrations. Error bars are one standard deviation.
Conclusions
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Batch cultivation in the different media resulted in peak cell concentrations from 2.5 × 106 to 9.0 × 106 cells/mL and a corresponding antibody titer from 220 to 860 mg/L. ActiCHO P and BalanCD CHO performed best in these cultures.
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Fed-batch cultivations substantially improved cell and product concentration. Feeding cultures in CD OptiCHO with EfficientFeed A and FunctionMAX or with Feed A and Feed B resulted in similar antibody concentrations and roughly doubled the antibody production compared to feeding with EfficientFeed A only.
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The highest titer was achieved in ActiCHO P in combination with Feed A and Feed B. In this medium a 6.3-fold improvement, compared with the previous batch cultivation, was observed. Further addition of FunctionMAX to these cultures did not significantly improve the antibody production.
Authors’ Affiliations
References
- Barrett S, Boniface R, Dhulipala P, Slade P, Tennico Y, Stramaglia M, Lio P, Gorfien S: Attaining Next-Level Titers in CHO Fed-Batch Cultures. BioProcess International. 2012, 10: 56-62.Google Scholar
Copyright
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.