The role of protein hydrolysates in prolonging viability and enhancing antibody production of CHO cells

Obaidi, Ismael; Mota, Letícia Martins; Quigley, Andrew; Butler, Michael

doi:10.1007/s00253-021-11244-8

Cited by 12 publications

(8 citation statements)

References 66 publications

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“…This suggests that yeast hydrolysate would be a good alternative to animal-based hydrolysate if the quality of the protein produced is the most important attribute. In a more recent study, cottonseed-derived hydrolysate enhances the galactosylation of CHO-S-RTX and CHO-EG2 cells, which in turn improves the product quality (Obaidi et al 2021 ). Furthermore, the glycosylation profile of the protein produced by CHO cells is equivalent between yeast and animal-based hydrolysate (Spearman et al 2014 ).…”

Section: The Use Of Hydrolysate In Animal Cell Culturementioning

confidence: 99%

Applications and analysis of hydrolysates in animal cell culture

Lim

et al. 2021

Bioresour. Bioprocess.

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Animal cells are used in the manufacturing of complex biotherapeutic products since the 1980s. From its initial uses in biological research to its current importance in the biopharmaceutical industry, many types of culture media were developed: from serum-based media to serum-free to protein-free chemically defined media. The cultivation of animal cells economically has become the ultimate goal in the field of biomanufacturing. Serum serves as a source of amino acids, lipids, proteins and most importantly growth factors and hormones, which are essential for many cell types. However, the use of serum is unfavorable due to its high price tag, increased lot-to-lot variations and potential risk of microbial contamination. Efforts are progressively being made to replace serum with recombinant proteins such as growth factors, cytokines and hormones, as well as supplementation with lipids, vitamins, trace elements and hydrolysates. While hydrolysates are more complex, they provide a diverse source of nutrients to animal cells, with potential beneficial effects beyond the nutritional value. In this review, we discuss the use of hydrolysates in animal cell culture and briefly cover the composition of hydrolysates, mode of action and potential contaminants with some perspectives on its potential role in animal cell culture media formulations in the future.

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Section: The Use Of Hydrolysate In Animal Cell Culturementioning

confidence: 99%

Applications and analysis of hydrolysates in animal cell culture

Lim

et al. 2021

Bioresour. Bioprocess.

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“…Due to the increasing demand for tools to monitor metabolites, a range of commercial bioprocess analyzers has been developed in the last couple of decades. Some of these are based on enzymatic analysis—such as the Cedex Bio® Analyzer (Roche) and the BioProfile FLEX2 (Nova biomedical) (Morris et al, 2021; Obaidi et al, 2021)—while others use the so‐called “miniaturized” MS analyzers—such as the MiD (Microsaic) and the Rebel (908 devices) (Hamilton et al, 2014; Synoground et al, 2021), employing a low‐resolution quadrupole and ion trap mass analyzers, respectively (Blakeman & Miller, 2021; Hemida et al, 2021). However, these analyzers are still almost exclusively being used at‐line or off‐line—thus limiting their monitoring potential—and only targeting the predefined set of compounds dictated by the vendor reagents.…”

Section: Introductionmentioning

confidence: 99%

On‐line untargeted metabolomics monitoring of an Escherichia coli succinate fermentation process

Cortada-Garcia

Haggarty

Moses

et al. 2022

Biotech & Bioengineering

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The real-time monitoring of metabolites (RTMet) is instrumental for the industrial production of biobased fermentation products. This study shows the first application of untargeted on-line metabolomics for the monitoring of undiluted fermentation broth samples taken automatically from a 5 L bioreactor every 5 min via flow injection mass spectrometry. The travel time from the bioreactor to the mass spectrometer was 30 s. Using mass spectrometry allows, on the one hand, the direct monitoring of targeted key process compounds of interest and, on the other hand, provides information on hundreds of additional untargeted compounds without requiring previous calibration data. In this study, this technology was applied in an Escherichia coli succinate fermentation process and 886 different m/z signals were monitored, including key process compounds (glucose, succinate, and pyruvate), potential biomarkers of biomass formation such as (R)-2,3-dihydroxy-isovalerate and (R)-2,3-dihydroxy-3-methylpentanoate and compounds from the pentose phosphate pathway and nucleotide metabolism, among others. The main advantage of the RTMet technology is that it allows the monitoring of hundreds of signals without the requirement of developing partial least squares regression models, making it a perfect tool for bioprocess monitoring and for testing many different strains and process conditions for bioprocess development.

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“…[1] Improving the production of recombinant proteins, including monoclonal antibodies (mAb), is critical for the biopharmaceutical industry. [2,3] There have been multiple attempts to increase the productivity of CHO cells by vector modification and cell engineering. [4][5][6] Genetic engineering targeting the cell cycle and autophagy pathways to regulate cell death in CHO cell cultures has received attention for enhancing the production of therapeutic proteins.…”

Section: Introductionmentioning

confidence: 99%

Apilimod enhances specific productivity in recombinant CHO cells through cell cycle arrest and mediation of autophagy

Xiao

Feng

et al. 2022

Biotechnology Journal

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Chinese hamster ovary (CHO) cells are expected to acquire the ability to produce higher recombinant therapeutic protein levels using various strategies. Genetic engineering targeting the cell cycle and autophagy pathways in the regulation of cell death in CHO cell cultures has received attention for enhancing the production of therapeutic proteins. In this study, we examined the small-molecule compound apilimod, which was found to have a positive influence on recombinant protein expression in CHO cells. This was confirmed by selective blocking of the cell cycle at the G0/G1 phase. Apilimod treatment resulted in decreased expression of cyclin-dependent kinase 3 (CDK3) and Cyclin C and increased expression of cyclin-dependent kinase suppressor p27Kip1, which are critical regulators of G1 cell cycle progression and important targets controlling cell proliferation. Furthermore, total transcription factor EB (TFEB) was lower in apilimod-treated CHO cells than in control cells, resulting in decreased lysosome biogenesis and autophagy with apilimod treatment. These multiple effects demonstrate the potential of apilimod for development as a novel enhancer for the production of recombinant proteins in CHO cell engineering.

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The role of protein hydrolysates in prolonging viability and enhancing antibody production of CHO cells

Cited by 12 publications

References 66 publications

Applications and analysis of hydrolysates in animal cell culture

Applications and analysis of hydrolysates in animal cell culture

On‐line untargeted metabolomics monitoring of an Escherichia coli succinate fermentation process

Apilimod enhances specific productivity in recombinant CHO cells through cell cycle arrest and mediation of autophagy

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