Recent advances in CHO cell line development for recombinant protein production

Tihanyi, Borbála; Nyitray, László

doi:10.1016/j.ddtec.2021.02.003

Cited by 131 publications

(82 citation statements)

References 73 publications

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“…At present, antibody drug development has become one of the fastest growing, most profitable, and most feasible biopharmaceutical fields in the pharmaceutical industry; in particular, monoclonal antibodies have become one of the most important therapeutic recombinant antibodies in the global pharmaceutical market ( Masuda et al, 2021 ; Savizi et al, 2021 ). Up to now, more than 300 biological drugs have been approved by the U.S. Food and Drug Administration (FDA), among which monoclonal antibodies are developing rapidly ( Tihanyi and Nyitray, 2020 ). The production of recombinant antibody drugs is mostly achieved by constructing expression vectors in vitro through genetic engineering technology.…”

Section: Introductionmentioning

confidence: 99%

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Zhang

Shan

Liang

et al. 2022

Front. Bioeng. Biotechnol.

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Recombinant antibodies are rapidly developing therapeutic agents; approximately 40 novel antibody molecules enter clinical trials each year, most of which are produced from Chinese hamster ovary (CHO) cells. However, one of the major bottlenecks restricting the development of antibody drugs is how to perform high-level expression and production of recombinant antibodies. The high-efficiency expression and quality of recombinant antibodies in CHO cells is determined by multiple factors. This review provides a comprehensive overview of several state-of-the-art approaches, such as optimization of gene sequence of antibody, construction and optimization of high-efficiency expression vector, using antibody expression system, transformation of host cell lines, and glycosylation modification. Finally, the authors discuss the potential of large-scale production of recombinant antibodies and development of culture processes for biopharmaceutical manufacturing in the future.

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Section: Introductionmentioning

confidence: 99%

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Zhang

Shan

Liang

et al. 2022

Front. Bioeng. Biotechnol.

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“…Around 70% of biopharmaceuticals are produced in Chinese hamster ovary (CHO) cells, with high productivities ranging between 0.1 and 10 g/L ( Tihanyi and Nyitray, 2020 ). One of the bottlenecks of plant-based production is the low yield of the recombinant protein ( Schillberg and Finnern, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Process Engineering of Biopharmaceutical Production in Moss Bioreactors via Model-Based Description and Evaluation of Phytohormone Impact

Ruiz-Molina

Parsons

Schroeder

et al. 2022

Front. Bioeng. Biotechnol.

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The moss Physcomitrella is an interesting production host for recombinant biopharmaceuticals. Here we produced MFHR1, a synthetic complement regulator which has been proposed for the treatment of diseases associated to the complement system as part of human innate immunity. We studied the impact of different operation modes for the production process in 5 L stirred-tank photobioreactors. The total amount of recombinant protein was doubled by using fed-batch or batch compared to semi-continuous operation, although the maximum specific productivity (mg MFHR1/g FW) increased just by 35%. We proposed an unstructured kinetic model which fits accurately with the experimental data in batch and semi-continuous operation under autotrophic conditions with 2% CO2 enrichment. The model is able to predict recombinant protein production, nitrate uptake and biomass growth, which is useful for process control and optimization. We investigated strategies to further increase MFHR1 production. While mixotrophic and heterotrophic conditions decreased the MFHR1-specific productivity compared to autotrophic conditions, addition of the phytohormone auxin (NAA, 10 µM) to the medium enhanced it by 470% in shaken flasks and up to 230% and 260%, in batch and fed-batch bioreactors, respectively. Supporting this finding, the auxin-synthesis inhibitor L-kynurenine (100 µM) decreased MFHR1 production significantly by 110% and 580% at day 7 and 18, respectively. Expression analysis revealed that the MFHR1 transgene, driven by the Physcomitrella actin5 (PpAct5) promoter, was upregulated 16 h after NAA addition and remained enhanced over the whole process, whereas the auxin-responsive gene PpIAA1A was upregulated within the first 2 hours, indicating that the effect of auxin on PpAct5 promoter-driven expression is indirect. Furthermore, the day of NAA supplementation was crucial, leading to an up to 8-fold increase of MFHR1-specific productivity (0.82 mg MFHR1/g fresh weight, 150 mg accumulated over 7 days) compared to the productivity reported previously. Our findings are likely to be applicable to other plant-based expression systems to increase biopharmaceutical production and yields.

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“…However, over the last 20 years an increase in process yields from 0.5 to 9 g/L was achieved [ 2 ]. Moreover, innovations in the manufacturing processes also aim to achieve product stability and quality along with low manufacturing costs, in reduced times [ 3 ].…”

Section: Introductionmentioning

confidence: 99%

“…Indeed, about 70% of the marketed biopharmaceutical proteins, and almost all mAbs, were produced in CHO cells [ 30 , 31 ]. Their popularity lies in their high productivity, consistent growth phenotypes, suitability for large-scale industrial culturing, ease of adaptation to various chemically defined media, lower susceptibility to infection by human viruses, and capability to perform human compatible glycosylation [ 3 ]. The HEK-293 cell line has been extensively used for the production of viral vectors for gene and cell therapy, and has also been validated as an efficient platform for the large-scale production of recombinant proteins due to its high transfectivity, rapid growth rate, and ability to grow in a serum-free, suspension culture [ 32 , 33 ].…”

Section: Introductionmentioning

confidence: 99%

Generation and Characterization of Stable Redox-Reporter Mammalian Cell Lines of Biotechnological Relevance

Perelmuter

Tiscornia

Comini

et al. 2022

Sensors

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Cellular functions such as DNA replication and protein translation are influenced by changes in the intracellular redox milieu. Exogenous (i.e., nutrients, deterioration of media components, xenobiotics) and endogenous factors (i.e., metabolism, growth) may alter the redox homeostasis of cells. Thus, monitoring redox changes in real time and in situ is deemed essential for optimizing the production of recombinant proteins. Recently, different redox-sensitive variants of green fluorescent proteins (e.g., rxYFP, roGFP2, and rxmRuby2) have been engineered and proved suitable to detect, in a non-invasive manner, perturbations in the pool of reduced and oxidized glutathione, the major low molecular mass thiol in mammals. In this study, we validate the use of cytosolic rxYFP on two cell lines widely used in biomanufacturing processes, namely, CHO-K1 cells expressing the human granulocyte macrophage colony-stimulating factor (hGM-CSF) and HEK-293. Flow cytometry was selected as the read-out technique for rxYFP signal given its high-throughput and statistical robustness. Growth kinetics and cellular metabolism (glucose consumption, lactate and ammonia production) of the redox reporter cells were comparable to those of the parental cell lines. The hGM-CSF production was not affected by the expression of the biosensor. The redox reporter cell lines showed a sensitive and reversible response to different redox stimuli (reducing and oxidant reagents). Under batch culture conditions, a significant and progressive oxidation of the biosensor occurred when CHO-K1-hGM-CSF cells entered the late-log phase. Medium replenishment restored, albeit partially, the intracellular redox homeostasis. Our study highlights the utility of genetically encoded redox biosensors to guide metabolic engineering or intervention strategies aimed at optimizing cell viability, growth, and productivity.

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Recent advances in CHO cell line development for recombinant protein production

Cited by 131 publications

References 73 publications

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Process Engineering of Biopharmaceutical Production in Moss Bioreactors via Model-Based Description and Evaluation of Phytohormone Impact

Generation and Characterization of Stable Redox-Reporter Mammalian Cell Lines of Biotechnological Relevance

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