Understanding the enhanced effect of dimethyl sulfoxide on hepatitis B surface antigen expression in the culture of Chinese hamster ovary cells on the basis of proteome analysis

Li, Junhui; Huang, Zhenyu; Sun, Xiangming; Yang, Pengyuan; Zhang, Yuanxing

doi:10.1016/j.enzmictec.2005.05.016

Cited by 21 publications

(12 citation statements)

References 28 publications

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“…For example, DMSO downregulates HSP70 and ERP57, which play an important role in stabilizing the folding and assembly intermediates of newly synthesized polypeptides. 20 The same authors also found that protein disulfide-isomerase was downregulated by DMSO, which regulated protein post-translation modifications and secretion pathway. 20 If a newly synthesized protein does not obtain the appropriate modifications, the protein may be degraded in Golgi body and not secreted outside of cells.…”

Section: Purification and Bioactivity Assay Of The Fusion Proteinmentioning

confidence: 91%

Production and characterization of active recombinant interleukin‐12/eGFP fusion protein in stably‐transfected DF1 chicken cells

Chen

Shen

et al. 2015

Biotechnology Progress

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The adjuvant activity of chicken interleukin-12 (chIL-12) protein has been described as similar to that of mammalian IL-12. Recombinant chIL-12 can be produced using several methods, but chIL-12 production in eukaryotic cells is lower than that in prokaryotic cells. Stimulating compounds, such as dimethyl sulfoxide (DMSO), can be added to animal cell cultures to overcome this drawback. In this study, we constructed a cell line, DF1/chIL-12 which stably expressed a fusion protein, chIL-12 and enhanced green fluorescent protein (eGFP) connected by a (G4 S)3 linker sequence. Fusion protein production was increased when cells were cultured in the presence of DMSO. When 1 × 10(6) DF1/chIL-12 cells were inoculated in a T-175 flask containing 30 mL of media, incubated for 15 h, and further cultivated in the presence of 4% DMSO for 48 h, the production of total fusion protein was mostly enhanced compared with the production of total fusion protein by using cell lysates induced with DMSO at other concentrations. The concentrations of the unpurified and purified total fusion proteins in cell lysates were 2,781 ± 2.72 ng mL(-1) and 2,207 ± 3.28 ng mL(-1) , respectively. The recovery rate was 79%. The fusion protein stimulated chicken splenocytes to produce IFN-γ, which was measured using an enzyme-linked immunosorbent assay, in the culture supernatant, indicating that treating DF1/chIL-12 cells with DMSO or producing chIL-12 in a fusion protein form does not have adverse effects on the bioactivity of chIL-12.

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Section: Purification and Bioactivity Assay Of The Fusion Proteinmentioning

confidence: 91%

Production and characterization of active recombinant interleukin‐12/eGFP fusion protein in stably‐transfected DF1 chicken cells

Chen

Shen

et al. 2015

Biotechnology Progress

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“…S protein of HBsAg contains 16 Cys residues, which participate in an extensive disulfide cross-linking within the assembled particle. Li et al (2006b) also found that protein disulfide-isomerase was downregulated by DMSO, which plays a very important role in protein posttranslational modifications and secretion pathway. If nascent protein does not get the appropriate modifications such as glycosylation and formation of disulfide bonds, the protein would be degraded in Golgi body and not secreted outside of cells.…”

Section: Discussionmentioning

confidence: 92%

“…This assembly process is the ratelimiting step in HBsAg particle secretion. Li et al (2006b) showed in proteome analysis of DMSO effect that the chaperone proteins such as HSP70 and ERP57 were downregulated by DMSO, which play an important role in stabilizing the folding and assembly intermediates of newly synthesized polypeptides and in glycoprotein folding in combination with calnexin and calreticulin. S protein of HBsAg contains 16 Cys residues, which participate in an extensive disulfide cross-linking within the assembled particle.…”

Section: Discussionmentioning

confidence: 98%

Gene transcription acceleration: main cause of hepatitis B surface antigen production improvement by dimethyl sulfoxide in the culture of Chinese hamster ovary cells

Wang

Zhang

2006

Biotech & Bioengineering

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The production and specific productivity of hepatitis B surface antigen (HBsAg) in recombinant Chinese hamster ovary (CHO) cells were increased by 81% and threefold, respectively, when supplemented with 1.5% dimethyl sulfoxide (DMSO) in the culture medium. To investigate the mechanism of DMSO effect on HBsAg production improvement, HBsAg mRNA level was measured by real-time PCR. HBsAg mRNA was increased by about 1.5-fold at 1.5% DMSO. The increase could derive from the increase of HBsAg gene copy number, the improvement of HBsAg mRNA stability, or the acceleration of HBsAg gene transcription. It was found that HBsAg gene copy number was not significantly changed in the cells stimulated with DMSO. HBsAg mRNA stability of cells with DMSO treatment was also not obviously different from control, and the mRNA half-life of 5.58 h in the cells at 1.5% DMSO was comparable to that of 5.36 h in the control culture. DMSO resulted in 80% increase in HBsAg gene transcription activity assessed using a nuclear run-on transcription assay. It could be deduced that the acceleration of HBsAg gene transcription is the main cause of HBsAg production improvement.

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“…Li et al (2006) investigated the effect of DMSO on increased productivity of hepatitis B surface antigen expression in recombinant CHO cells using differential protein profiling using a 2D-PAGE and MS-based approach. They identified seven proteins whose expression was significantly altered following exposure to DMSO and an associated increase in productivity.…”

Section: Media Supplementationmentioning

confidence: 99%

Proteomic profiling of recombinant cells from large-scale mammalian cell culture processes

Meleady

2007

Cytotechnology

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Global expression profiling of mammalian cells used for the production of biopharmaceuticals will allow greater insights into the molecular mechanisms that result in a high producing cellular phenotype. These studies may give insights for genetic intervention to possibly create better host cell lines or even to provide clues to more rational strategies for cell line and process development. In this review I will focus on the contribution of proteomic technologies to a greater understanding of the biology of Chinese hamster ovary cells and other producing cell lines such as NS0 mouse cells.

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Understanding the enhanced effect of dimethyl sulfoxide on hepatitis B surface antigen expression in the culture of Chinese hamster ovary cells on the basis of proteome analysis

Cited by 21 publications

References 28 publications

Production and characterization of active recombinant interleukin‐12/eGFP fusion protein in stably‐transfected DF1 chicken cells

Production and characterization of active recombinant interleukin‐12/eGFP fusion protein in stably‐transfected DF1 chicken cells

Gene transcription acceleration: main cause of hepatitis B surface antigen production improvement by dimethyl sulfoxide in the culture of Chinese hamster ovary cells

Proteomic profiling of recombinant cells from large-scale mammalian cell culture processes

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