Applications of small molecules in modulating productivity and product quality of recombinant proteins produced using cell cultures

Xu, Ping; Xu, Sen; He, Chunyan; Khetan, Anurag

doi:10.1016/j.biotechadv.2020.107577

Cited by 9 publications

(2 citation statements)

References 106 publications

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“…Over the last few decades, a series of process and cell engineering approaches have increased CHO cell-specific productivity (extensively reviewed by Fischer et al, 2015;O'Flaherty et al 2020;Xu et al, 2020). At the process level, strategies based on cell proliferation control, such as low temperature or sodium butyrate (NaBu) treatment, have been successful in enhancing cell-specific productivities (q P ) in CHO cell cultures.…”

Section: Introductionmentioning

confidence: 99%

Combined gene and environmental engineering offers a synergetic strategy to enhance r‐protein production in Chinese hamster ovary cells

Torres

Dickson

2021

Biotech & Bioengineering

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Environmental growth-inhibition conditions (GICs) have been used extensively for increasing cell-specific productivity (q P ) of Chinese hamster ovary (CHO) cells, with the most common being temperature downshift and sodium butyrate (NaBu) treatment. B lymphocyte-induced maturation protein-1 (BLIMP1) overexpression in CHO cells can also inhibit cell growth and increase product titers and q P . Given the similar responses, this study evaluated the individual and combined effects of BLIMP1 expression, low temperature, and NaBu treatment on culture performance, cell metabolism, and recombinant protein production of CHO cells. As expected, all three interventions decreased cell growth, arrested cells in G1/G0 cell cycle phase, and increased q P . However, CHO cells presented different responses when considering cell viability, recombinant gene expression, and cell metabolism that indicated differences in the molecular loci by which BLIMP1 and GICs generated higher productivities. Combinations of BLIMP1 expression and GICs acted synergistically to inhibit cell growth and maximize r-protein production, with the BLIMP1/NaBu condition leading to the most significant improvements in product titers and q P . This latter condition also proved to substantially increase product yields (up to 9.8 g immunoglobulin G1 [IgG1]/L and 2.2 g erythropoietin-Fc [EPO-Fc]/L) and q P (up to 179 pg/cell/day [pcd] for IgG1 and 30 pcd for EPO-Fc) in highdensity perfusion cultures. These findings offered mechanistic insights about the productivity-enhancing effects of BLIMP1 and GICs, as well as their complementarity for generating highly productive processes.

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Section: Introductionmentioning

confidence: 99%

Combined gene and environmental engineering offers a synergetic strategy to enhance r‐protein production in Chinese hamster ovary cells

Torres

Dickson

2021

Biotech & Bioengineering

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“…Mammalian cells have been used for decades to produce a variety of recombinant proteins (Xu, Xu et al 2020), but the low yield of transient gene expression in mammalian cells has limited their use. Although much work has been published to characterize promoters, enhancers, and PTREs for this purpose, gene expression involves the integration of numerous different signals affecting different regulatory elements, and there are many possible functional interactions between regulatory elements (Bergman, Jones et al 2022, Martinez-Ara, Comoglio et al 2022).…”

Section: Discussionmentioning

confidence: 99%

Comparison of the effects of different combinations of expression regulatory elements on gene expression in mammalian cells

Zhou

Zhang

et al. 2022

Preprint

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Background At present, the function of numerous individual regulatory elements has been characterized extensively, but the combined effect of various cis-regulatory elements on gene expression in mammalian cells has been less discussed. Results In the present study, we examined the function of the combined effect of various cis-regulatory elements on gene expression in mammalian cells. To detect the function of different combinations of cis-acting elements on gene expression, we compared the effects of four promoters (PGK, Polr2a and EF-1α core promoter), two enhancers (CMV and SV40 enhancer), two introns (EF-1α intron A and hybrid intron), two terminators (CYC1 and TEF terminator) and their different combinations on gene expression. We constructed expression vectors with different combinations of these regulatory elements and detect the expression of eGFP by fluorescence intensity contrast and western blotting. Here we report that different combinations of cis-acting elements have a significant effect on protein expression. The vector with CMV enhancer, EF-1α core promoter and TEF terminator can express about three times higher eGFP than the unmodified vector in different animal cells. Conclusions We demonstrate that a direct combination of multiple regulatory elements capable of regulating gene expression did not exhibit synergistic effects. Our results show that regulation of protein expression can be achieved by optimizing the combination of cis-acting elements, and these findings can be used in biological applications that require regulation of gene expression.

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Effects and mechanisms of animal-free hydrolysates on recombination protein yields in CHO cells

Zhang

Wang

et al. 2022

Appl Microbiol Biotechnol

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Applications of small molecules in modulating productivity and product quality of recombinant proteins produced using cell cultures

Cited by 9 publications

References 106 publications

Combined gene and environmental engineering offers a synergetic strategy to enhance r‐protein production in Chinese hamster ovary cells

Combined gene and environmental engineering offers a synergetic strategy to enhance r‐protein production in Chinese hamster ovary cells

Comparison of the effects of different combinations of expression regulatory elements on gene expression in mammalian cells

Effects and mechanisms of animal-free hydrolysates on recombination protein yields in CHO cells

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