Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Ritacco, Frank V.; Wu, Yongqi; Khetan, Anurag

doi:10.1002/btpr.2706

Cited by 183 publications

(164 citation statements)

References 212 publications

(429 reference statements)

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“…Notably, tremendous research efforts are required for media composition optimization ( Hunter et al, 2019 ). The conventional approach is focused on the development of an optimized cultivation media via component titration using the “one factor at a time” (OFAT) method ( Ritacco et al, 2018 ). Recently, by understanding how certain intracellular anabolic pathways contribute to protein expression, there is considerable interest in improving protein production efficiency by tuning host cell metabolism via incorporation of expression pathway inducers into the cultivation media ( Huang et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Chlorella vulgaris Extract as a Serum Replacement That Enhances Mammalian Cell Growth and Protein Expression

Chua

Zhang³

et al. 2020

Front. Bioeng. Biotechnol.

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The global cell culture market is experiencing significant growth due to the rapid advancement in antibody-based and cell-based therapies. Both rely on the capacity of different living factories, namely prokaryotic and eukaryotic cells, plants or animals for reliable and mass production. The ability to improve production yield is of important concern. Among many strategies pursued, optimizing the complex nutritional requirements for cell growth and protein production has been frequently performed via culture media component titration and serum replacement. The addition of specific ingredients into culture media to modulate host cells' metabolism has also recently been explored. In this study, we examined the use of extracted bioactive components of the microalgae Chlorella vulgaris, termed chlorella growth factor (CGF), as a cell culture additive for serum replacement and protein expression induction. We first established a chemical fingerprint of CGF using ultraviolet-visible spectroscopy and liquid chromatography-mass spectrometry and evaluated its ability to enhance cell proliferation in mammalian host cells. CGF successfully promoted the growth of Chinese hamster ovary (CHO) and mesenchymal stem cells (MSC), in both 2D and 3D cell cultures under reduced serum conditions for up to 21 days. In addition, CGF preserved cell functions as evident by an increase in protein expression in CHO cells and the maintenance of stem cell phenotype in MSC. Taken together, our results suggest that CGF is a viable culture media additive and growth matrix component, with wide ranging applications in biotechnology and tissue engineering.

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Section: Introductionmentioning

confidence: 99%

Chlorella vulgaris Extract as a Serum Replacement That Enhances Mammalian Cell Growth and Protein Expression

Chua

Zhang³

et al. 2020

Front. Bioeng. Biotechnol.

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“…Reproducible and scalable media preparation is a challenging task . Media preparation processes can vary in their specific preparation parameters dependent from scale and type of preparation vessel.…”

Section: Resultsmentioning

confidence: 99%

“…Besides analytical and biological variability, a major source for variable process performances is often reported to be linked to the chemical media composition . Cell culture media often consist of more than 50 components, including substances with known stability issues such as cysteine, insulin, and riboflavin . Quality control for cell culture media may include raw material screening with spectroscopic methods as near‐infrared and mid‐infrared (MIR) spectroscopy .…”

Section: Introductionmentioning

confidence: 99%

Towards robust cell culture processes — Unraveling the impact of media preparation by spectroscopic online monitoring

Brunner

Brosig

Losing

et al. 2019

Engineering in Life Sciences

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Biopharmaceutical manufacturing processes can be affected by variability in cell culture media, e.g. caused by raw material impurities. Although efforts have been made in industry and academia to characterize cell culture media and raw materials with advanced analytics, the process of industrial cell culture media preparation itself has not been reported so far. Within this publication, we first compare mid‐infrared and two‐dimensional fluorescence spectroscopy with respect to their suitability as online monitoring tools during cell culture media preparation, followed by a thorough assessment of the impact of preparation parameters on media quality. Through the application of spectroscopic methods, we can show that media variability and its corresponding root cause can be detected online during the preparation process. This methodology is a powerful tool to avoid batch failure and is a valuable technology for media troubleshooting activities. Moreover, in a design of experiments approach, including additional liquid chromatography–mass spectrometry analytics, it is shown that variable preparation parameters such as temperature, power input and preparation time can have a strong impact on the physico‐chemical composition of the media. The effect on cell culture process performance and product quality in subsequent fed‐batch processes was also investigated. The presented results reveal the need for online spectroscopic methods during the preparation process and show that media variability can already be introduced by variation in media preparation parameters, with a potential impact on scale‐up to a commercial manufacturing process.

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“…Conventional and emerging technologies for the expression of recombinant multi-protein complexes in mammalian expression systems were summarized in a review (Baser and van den Heuvel, 2016). The evolution of culture media, nutrient composition and formulation needs, optimization strategies, consistency and scalability of powder and liquid media preparation for industrial applications, and key recent advances driving progress in CHO cell culture medium design and development have been described (Ritacco et al, 2018). The major technological advancements along with the areas of application of CHO cell line development and engineering were discussed by Hong et al (2018a).…”

Section: Mammalian Cellsmentioning

confidence: 99%

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

Tripathi

Shrivastava

2019

Front. Bioeng. Biotechnol.

369

243

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Infectious diseases, along with cancers, are among the main causes of death among humans worldwide. The production of therapeutic proteins for treating diseases at large scale for millions of individuals is one of the essential needs of mankind. Recent progress in the area of recombinant DNA technologies has paved the way to producing recombinant proteins that can be used as therapeutics, vaccines, and diagnostic reagents. Recombinant proteins for these applications are mainly produced using prokaryotic and eukaryotic expression host systems such as mammalian cells, bacteria, yeast, insect cells, and transgenic plants at laboratory scale as well as in large-scale settings. The development of efficient bioprocessing strategies is crucial for industrial production of recombinant proteins of therapeutic and prophylactic importance. Recently, advances have been made in the various areas of bioprocessing and are being utilized to develop effective processes for producing recombinant proteins. These include the use of high-throughput devices for effective bioprocess optimization and of disposable systems, continuous upstream processing, continuous chromatography, integrated continuous bioprocessing, Quality by Design, and process analytical technologies to achieve quality product with higher yield. This review summarizes recent developments in the bioprocessing of recombinant proteins, including in various expression systems, bioprocess development, and the upstream and downstream processing of recombinant proteins.

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Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Cited by 183 publications

References 212 publications

Chlorella vulgaris Extract as a Serum Replacement That Enhances Mammalian Cell Growth and Protein Expression

Chlorella vulgaris Extract as a Serum Replacement That Enhances Mammalian Cell Growth and Protein Expression

Towards robust cell culture processes — Unraveling the impact of media preparation by spectroscopic online monitoring

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

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