Erythropoietin production from CHO cells grown by continuous culture in a fluidized‐bed bioreactor

Wang, M-D.; Yang, Ming; Huzel, N.; Butler, Michael

doi:10.1002/bit.10144

Cited by 73 publications

(53 citation statements)

References 46 publications

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“…During the development of a perfusion process producing EPO, it was also observed that a constant perfusion rate allowed the system to approach steady state, in which the concentration of the main components were maintained at a constant level (Wang et al 2002).…”

Section: Discussionmentioning

confidence: 99%

“…In the course of reducing the perfusion rate, several studies were conducted where the concentration of glucose (Dowd et al 2001;Wang et al 2002) is used as an indicator of other nutrients level in the feed medium in order to operate the bioreactor at a low perfusion rate without accumulating in the culture high levels of toxic byproducts such as lactate and ammonia Sugiura and Kakuzaki 1998). Modification of culture parameters such as pH or temperature (Chuppa et al 1997) is also a common strategy to optimize culture conditions and reduce medium perfusion needs.…”

Section: Introductionmentioning

confidence: 99%

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Optimization of the medium perfusion rate in a packed-bed bioreactor charged with CHO cells

2004

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In the present study, the optimal medium perfusion rate to be used for the continuous culture of a recombinant CHO cell line in a packed-bed bioreactor made of Fibra-Cel Ò disk carriers was determined. A first-generation process had originally been designed with a high perfusion rate, in order to rapidly produce material for pre-clinical and early clinical trials. It was originally operated with a perfusion of 2.6 vvd during production phase in order to supply the high cell density ($2.5 · 10 7 cell ml À1 of packed-bed) with sufficient fresh medium. In order to improve the economics of this process, a reduction of the medium perfusion rate by À25% and À50% was investigated at small-scale. The best option was then implemented at pilot scale in order to further produce material for clinical trials with an improved second-generation process. With a À25% reduction of the perfusion rate, the volumetric productivity was maintained compared to the first-generation process, but a À30% loss of productivity was obtained when the medium perfusion rate was further reduced to À50% of its original level. The protein quality under reduced perfusion rate conditions was analyzed for purity, N-glycan sialylation level, abundance of dimers or aggregates, and showed that the quality of the final drug substance was comparable to that obtained in reference conditions. Finally, a reduction of À25% medium perfusion was implemented at pilot scale in the second-generation process, which enabled to maintain the same productivity and the same quality of the molecule, while reducing costs of media, material and manpower of the production process. For industrial applications, it is recommended to test whether and how far the perfusion rate can be decreased during the production phase -provided that the product is not sensitive to residence time -with the benefits of reduced cost of goods and to simplify manufacturing operations.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimization of the medium perfusion rate in a packed-bed bioreactor charged with CHO cells

2004

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“…Butyrate treatment in CHO cells affects cellular processes such as cytoskeleton reorganization, protein transport, nucleosome assembly, nucleotide metabolism, glycosylation, protein folding, oxidative stress responses, lipid metabolism, cholesterol biosynthesis, glycolysis, cell cycle progression, and apoptosis (Yee et al 2008;De Leon et al 2007). Butyrate has been used in cell engineering strategies, and butyrate treatment in CHO cells often increases recombinant protein expression (Chun et al 2003;De Leon et al 2007;Hendrick et al 2001;Kim and Lee 2000;Chotigeat et al 1994;Mimura et al 2001;Wang et al 2002;Jiang and Sharfstein 2008). The current work explores the effect of butyrate on glycosaminoglycan (GAG) profiles in recombinant CHO cells in our efforts to produce a bioengineered heparin (HP).…”

Section: Butyrate Work As a Histone Deacetylase Inhibitor Andmentioning

confidence: 99%

“…Sodium butyrate (CH 3 CH 2 CH 2 CO 2 -) is a histone deacetylase inhibitor that modulates gene expression of cellular processes, particularly transgene expression in Chinese hamster ovary (CHO) cells (Chotigeat et al 1994;Chun et al 2003;De Leon et al 2007;Hendrick et al 2001;Jiang and Sharfstein 2008;Kim and Lee 2000;Mimura et al 2001;Wang et al 2002).…”

Section: Introductionmentioning

confidence: 99%

Modulation of heparan sulfate biosynthesis by sodium butyrate in recombinant CHO cells

et al. 2014

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Sodium butyrate, a histone deacetylase inhibitor, has been used to improve transgene expression in Chinese hamster ovary (CHO) cells. The current study explores the impact of butyrate treatment on heparan sulfate (HS) biosynthesis and structural composition in a recombinant CHO-S cell line expressing enzymes in the heparin (HP)/(HS) biosynthetic pathway (Dual-10 stably expressing NDST2 and HS3st1). Flow cytometric analysis showed that antithrombin binding was increased in Dual-10 cells and basic fibroblast growth factor binding was decreased in response to sodium butyrate treatment. The results were in agreement with the AMAC-LCMS (2-aminoacridine-tagged HS/HP analysis by liquid chromatography mass spectrometry) data that showed that there was an increase in heparan sulfate tri-sulfated disaccharides and a decrease in N-sulfated disaccharides in the butyrate-treated cells. However, we could not detect any changes in the chondroitin sulfate pathway in Dual-10 cells treated with butyrate. The current study is the first to report the effect of butyrate on glycosaminoglycan profiles.

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“…Sodium butyrate는 가장 널리 사용되는 SME 중 하나 로, 동물세포배양을 이용하여 tPA [9], α 1 -antitrypsin [10], hEPO [11], scu-PA [12], immunoglobulin [13] Fig. 3(a) .…”

Section: 서론unclassified

Enhanced Production of hCTLA4Ig by Adding Sodium Butyrate and Sodium Pyruvate

유미희¹,

김수진²,

권준영³

et al. 2011

KSBB Journal

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1) Human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig), an immunosuppressive agent, was expressed in transgenic rice cells using RAmy3D promoter and RAmy1A signal peptide for the inducible production and secretion into culture media by sugar depletion. In this study, sodium butyrate was used as a small molecular enhancer (SME) to enhance the production of hCTLA4Ig in transgenic rice cell suspension cultures. When 1 mM sodium butyrate was added in sugar-free media, relative viability was not reduced, while the productivity was improved 1.3-fold. In addition, by supplementing 87 mM sodium pyruvate as an alternative energy source during the production phase, death rate of the cells was decreased. When sodium pyruvate was not added, most cells became dead at day 6. However, by adding sodium pyruvate, 18% of viability can be maintained until day 10 and the production of hCTLA4Ig was enhanced 1.4-fold. When the combination of sodium pyruvate and sodium butyrate at optimum concentrations was added, the highest viability and hCTLA4Ig production could be obtained.

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Erythropoietin production from CHO cells grown by continuous culture in a fluidized‐bed bioreactor

Cited by 73 publications

References 46 publications

Optimization of the medium perfusion rate in a packed-bed bioreactor charged with CHO cells

Optimization of the medium perfusion rate in a packed-bed bioreactor charged with CHO cells

Modulation of heparan sulfate biosynthesis by sodium butyrate in recombinant CHO cells

Enhanced Production of hCTLA4Ig by Adding Sodium Butyrate and Sodium Pyruvate

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