A reflection on the improvement of Chinese hamster ovary cell-based bioprocesses through advances in proteomic techniques

Nguyen, Melanie; Zimmer, Aline

doi:10.1016/j.biotechadv.2023.108141

Cited by 4 publications

(3 citation statements)

References 127 publications

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“…The quality of a therapeutic protein refers to its physical, chemical, and biological properties that enable them to function properly (Ha et al 2022 ). Quality properties such as glycosylation, aggregation, charge variant, and degradation determine the activity, efficiency, safety, pharmacokinetics, and pharmacodynamics of RTPs (Nguyen and Zimmer 2023 ; Torkashvand and Vaziri 2017 ). However, the biggest obstacles encountered when using mammalian cells such as CHO cell lines is the degradation of POIs by peptides expressed and secreted in the cell culture medium, which is also one of the important factors affecting the quality of antibodies (Wen et al 2018 ).…”

Section: Degradation Of Rtpsmentioning

confidence: 99%

“…Degradation, mainly includes fragmentation, hydrolysis, and cleavage, refers to the destruction of protein structure in the production of RTPs under the action of enzymes or non-enzymes, resulting in the heterogeneity of glycosylation, charge, etc., which are crucial to the stability, half-life, immunogenicity, and biological function of the drug (Nguyen and Zimmer 2023 ; Torkashvand and Vaziri 2017 ). The heterogeneity caused by degradation will also affect the yield of the obtained proteins, and also increase the burden on downstream purification processes (Bryan et al 2021 ; Budge et al 2021 ).…”

Section: Degradation Of Rtpsmentioning

confidence: 99%

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Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

Geng,

Zhao,

Zhang

et al. 2024

Appl Microbiol Biotechnol

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Mammalian cell lines are frequently used as the preferred host cells for producing recombinant therapeutic proteins (RTPs) having post-translational modified modification similar to those observed in proteins produced by human cells. Nowadays, most RTPs approved for marketing are produced in Chinese hamster ovary (CHO) cells. Recombinant therapeutic antibodies are among the most important and promising RTPs for biomedical applications. One of the issues that occurs during development of RTPs is their degradation, which caused by a variety of factors and reducing quality of RTPs. RTP degradation is especially concerning as they could result in reduced biological functions (antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity) and generate potentially immunogenic species. Therefore, the mechanisms underlying RTP degradation and strategies for avoiding degradation have regained an interest from academia and industry. In this review, we outline recent progress in this field, with a focus on factors that cause degradation during RTP production and the development of strategies for overcoming RTP degradation. Key points • The recombinant therapeutic protein degradation in CHO cell systems is reviewed. • Enzymatic factors and non-enzymatic methods influence recombinant therapeutic protein degradation. • Reducing the degradation can improve the quality of recombinant therapeutic proteins.

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Section: Degradation Of Rtpsmentioning

confidence: 99%

Section: Degradation Of Rtpsmentioning

confidence: 99%

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

Geng,

Zhao,

Zhang

et al. 2024

Appl Microbiol Biotechnol

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“…Chinese hamster ovary (CHO) cells have long been the cell line of choice for therapeutic recombinant protein expression, and approximately 70% of all globally approved therapeutic recombinant proteins have been produced in CHO cell lines ( Lalonde and Durocher, 2017 ; Dhara et al, 2018 ; Dahodwala and Lee, 2019 ). Over the past 2 decades, with the development of optimized media, process control, and cell engineering, mammalian cell production has made qualitative leaps, with CHO protein production exceeding 10 g/L by batch replenishment ( Kelley, 2009 ; Nguyen and Zimmer, 2023 ). However, longer cultivation times, lower product yield, and phenotypic heterogeneity of transgenic cell clones have been identified as limitations of mammalian cell lines compared with E. coli, yeast, and insect cell lines ( Dalton and Barton, 2014 ; Li et al, 2016 ; Wells and Robinson, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

Construction and application of a multifunctional CHO cell platform utilizing Cre/lox and Dre/rox site-specific recombination systems

Zhang,

Chang,

Miao

et al. 2023

Front. Bioeng. Biotechnol.

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During the development of traditional Chinese hamster ovary (CHO) cell lines, target genes randomly integrate into the genome upon entering the nucleus, resulting in unpredictable productivity of cell clones. The characterization and screening of high-yielding cell lines is a time-consuming and expensive process. Site-specific integration is recognized as an effective approach for overcoming random integration and improving production stability. We have designed a multifunctional expression cassette, called CDbox, which can be manipulated by the site-specific recombination systems Cre/lox and Dre/rox. The CDbox expression cassette was inserted at the Hipp11(H11) locus hotspot in the CHO-K1 genome using CRISPR/Cas9 technology, and a compliant CHO-CDbox cell platform was screened and obtained. The CHO-CDbox cell platform was transformed into a pool of EGFP-expressing cells using Cre/lox recombinase-mediated cassette exchange (RMCE) in only 2 weeks, and this expression remained stable for at least 75 generations without the need for drug stress. Subsequently, we used the Dre/rox system to directly eliminate the EGFP gene. In addition, two practical applications of the CHO-CDbox cell platform were presented. The first was the quick construction of the Pembrolizumab antibody stable expression strain, while the second was a protocol for the integration of surface-displayed and secreted antibodies on CHO cells. The previous research on site-specific integration of CHO cells has always focused on the single functionality of insertion of target genes. This newly developed CHO cell platform is expected to offer expanded applicability for protein production and gene function studies.

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Enhancing the productivity and proliferation of CHO-K1 cells by oncoprotein YAP (Yes-associated protein)

Roshanmehr,

Abdoli,

Bazi

et al. 2024

Appl Microbiol Biotechnol

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CHO cells are extensively employed in biological drug industry to manufacture therapeutic proteins. Nevertheless, production of biopharmaceuticals faces obstacles such as limited growth and inadequate productivity. Employing host cell engineering techniques for CHO cells serves as a valuable approach to address the constraints encountered in biologics manufacturing. Despite advancements, most techniques focus on specific genes to address individual cellular challenges. The significance of YAP, transcriptional co-activator, cannot be overstated due to its involvement in regulating organ size and tumor formation. YAP’s influence extends to various cellular processes and is regulated by kinase cascade in the Hippo pathway, which phosphorylates serine residues in specific LATS recognition motifs. Activation of YAP has been observed to impact both the size and quantity of cells. This research investigates the effects of YAP5SA on proliferation, apoptosis, and productivity in CHO-K1 cells. YAP5SA, with mutations in all five LATS-target sites, is selected for its heightened activity and resistance to repression through the Hippo-LATS1/2 kinase signaling pathway. Plasmid harboring YAP5SA was transfected into EPO-CHO and the influence of YAP5SA overexpression was investigated. According to our findings, transfection of EPO-CHO cells with YAP5SA exhibited a substantial enhancement in CHO cell productivity, resulting in a 3-fold increase in total protein and EPO, as well as a 1.5-fold increase in specific productivity. Additionally, it significantly contributes in augmenting viability, size, and proliferation. Overall, the findings of this study exemplify the potential of utilizing YAP5SA to impact particular cellular mechanisms, thereby presenting an avenue for customizing cells to fulfill production demands. Key points • YAP5SA in CHO cells boosts growth, reduces apoptosis, and significantly improves productivity. • YAP5SA regulates genes involved in proliferation, survival, and mTOR activation. • YAP5SA increases productivity by improving cell cycle, c-MYC expression, and mTOR pathway.

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A reflection on the improvement of Chinese hamster ovary cell-based bioprocesses through advances in proteomic techniques

Cited by 4 publications

References 127 publications

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

Recombinant therapeutic proteins degradation and overcoming strategies in CHO cells

Construction and application of a multifunctional CHO cell platform utilizing Cre/lox and Dre/rox site-specific recombination systems

Enhancing the productivity and proliferation of CHO-K1 cells by oncoprotein YAP (Yes-associated protein)

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