Enhanced Production of Monomeric Interferon-β by CHO Cells through the Control of Culture Conditions

Rodrı́guez, José F.; Spearman, Maureen; Huzel, N.; Butler, Michael

doi:10.1021/bp049807b

Cited by 137 publications

(118 citation statements)

References 48 publications

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“…DMSO treatment can have a stimulatory effect on protein expression in animal cells (14,15) while sodium butyrate has been known to increase recombinant protein expression in animal cells (16,17). Although the mechanism of DMSO and sodium butyrate-mediated induction of protein expression is not clearly defined, DMSO is suggested to improve recombinant protein production by increasing the stability of the recombinant protein or by increasing transcription efficiency a The content of recombinant VP1 obtained from medium fraction of non-treated cultures was represented as 100.…”

Section: Resultsmentioning

confidence: 99%

Enhancement of Protein Productivity of Recombinant Hepatitis A Virus VP1 in Stably TransfectedDrosophila melanogasterS2 Cells

et al. 2012

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The effect of DMSO and sodium butyrate on the production of recombinant hepatitis A virus (HAV) capsid protein VP1 was evaluated and optimized in the culture of stably transfected Drosophila melanogaster S2 cells using culture plates and spinner flasks. The effect of DMSO and sodium butyrate was also evaluated to improve the recombinant VP1 production in stably transfected Drosophila S2 cells. A production level of 0.88 mg of recombinant VP1/liter was obtained in the culture-plate culture of stably transfected S2 cells at 6 days after induction with 0.5 mM CuSO 4 . The supplements of 2% DMSO and 10 mM sodium butyrate at 4 days post-inoculation increased recombinant VP1 accumulation by 141 and 104%, respectively, resulting in 2.17 and 1.7 mg/liter of recombinant VP1 production. In spinner flasks, recombinant VP1 production reached maximum level at 9 days after induction with 0.5 mM CuSO 4 , with approximately 4.96 mg/liter of recombinant VP1 production level. When 2% DMSO or 10 mM sodium butyrate was added at 5 days post-inoculation, the recombinant VP1 production was increased to 8.35 and 5.85 mg/liter, respectively. However, the synergistic effects of DMSO and sodium butyrate were not observed. These results indicate that DMSO and/or sodium butyrate can be successfully used to improve the recombinant HAV VP1 production in culture plates and spinner flasks.

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Section: Resultsmentioning

confidence: 99%

Enhancement of Protein Productivity of Recombinant Hepatitis A Virus VP1 in Stably TransfectedDrosophila melanogasterS2 Cells

et al. 2012

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“…Consequently, its adoption at large scale is questioned by the inefficient metabolic utilization, but due to the development of acetylated ManNAc analogs, which are metabolized up to 900-fold more efficiently than their natural counterparts 138 , this strategy remains interesting to modulate sialylation. At a concentration range of 1-10% (v/v), glycerol (1,2,3-propanetriol) displayed an enhancing effect on the level of sialylation in IFN-β batch cultures 139 . Fluorinated sialic acid analogs readily cross the cell membrane and do not negatively impact cell growth and viability.…”

Section: Sialylationmentioning

confidence: 99%

“…It was reported that by reducing glutamine concentrations to 0 mM in CHO-K1 batch and perfusion cultures, sialylation was inhibited, thus leading to more abundant neutral N-linked glycans 117 . Dimethyl sulfoxide (DMSO) from 1 to 8% (v/v) reduced sialylation but at the same time negatively impacted cell proliferation 139 . Nucleotide-sugar precursors modulate intracellular nucleotide-sugar pools and the resulting sialylation and antennarity levels.…”

Section: Sialylationmentioning

confidence: 99%

“…Other compounds, such as DMSO (1-8%, v/v) and glycerol (1-2%, v/v), exhibited protein stabilization capabilities 139,173 . In contrast, the addition of copper into the cell culture medium could slightly increase aggregation, whereas a supplementation of the culture medium with cysteine known for to its mild reducing characteristics on disulfide bond bridges, resulted in a substantial decrease in aggregate content and a corresponding increase in single chain species 54 .…”

Section: Aggregatesmentioning

confidence: 99%

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Tailoring recombinant protein quality by rational media design

Brühlmann

Jordan

Hemberger³

et al. 2015

Biotechnology Progress

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Clinical efficacy and safety of recombinant proteins are closely associated with their structural characteristics. The major quality attributes comprise glycosylation, charge variants (oxidation, deamidation, and C‐ & N‐terminal modifications), aggregates, low‐molecular‐weight species (LMW), and misincorporation of amino acids in the protein backbone. Cell culture media design has a great potential to modulate these quality attributes due to the vital role of medium in mammalian cell culture. The purpose of this review is to provide an overview of the way both classical cell culture medium components and novel supplements affect the quality attributes of recombinant therapeutic proteins expressed in mammalian hosts, allowing rational and high‐throughput optimization of mammalian cell culture media. A selection of specific and/or potent inhibitors and activators of oligosaccharide processing as well as components affecting multiple quality attributes are presented. Extensive research efforts in this field show the feasibility of quality engineering through media design, allowing to significantly modulate the protein function. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:615–629, 2015

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“…Limitations in the secretion of recombinant proteins can impact both protein quality and yield, which can have a negative impact on downstream processes. Published reports have focused on the characterization of such ‘difficult‐to‐express’ recombinant proteins and described the modification of culture conditions 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or the design of appropriate cell/protein engineering strategies to overcome restrictions in their production 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26. However, little is known regarding the mechanisms underpinning poor recombinant protein production, particularly between proteins of high sequence similarity.…”

mentioning

confidence: 99%

A protein chimera strategy supports production of a model “difficult‐to‐express” recombinant target

et al. 2018

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Due in part to the needs of the biopharmaceutical industry, there has been an increased drive to generate high quality recombinant proteins in large amounts. However, achieving high yields can be a challenge as the novelty and increased complexity of new targets often makes them ‘difficult‐to‐express’. This study aimed to define the molecular features that restrict the production of a model ‘difficult‐to‐express’ recombinant protein, Tissue Inhibitor Metalloproteinase‐3 (TIMP‐3). Building from experimental data, computational approaches were used to rationalize the redesign of this recombinant target to generate a chimera with enhanced secretion. The results highlight the importance of early identification of unfavourable sequence attributes, enabling the generation of engineered protein forms that bypass ‘secretory’ bottlenecks and result in efficient recombinant protein production.

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Enhanced Production of Monomeric Interferon-β by CHO Cells through the Control of Culture Conditions

Cited by 137 publications

References 48 publications

Enhancement of Protein Productivity of Recombinant Hepatitis A Virus VP1 in Stably TransfectedDrosophila melanogasterS2 Cells

Enhancement of Protein Productivity of Recombinant Hepatitis A Virus VP1 in Stably TransfectedDrosophila melanogasterS2 Cells

Tailoring recombinant protein quality by rational media design

A protein chimera strategy supports production of a model “difficult‐to‐express” recombinant target

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