Fedbatch Culture and Dynamic Nutrient Feeding

Wlaschin, Katie F.; Hu, Wei Shou

doi:10.1007/10_015

Cited by 67 publications

(60 citation statements)

References 70 publications

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“…This network consists of glycolysis, tricarboxylic acid cycle (TCA), and amino acid metabolic pathways (Wlaschin and Hu 2006;Dean and Reddy 2013). The stoichiometry matrix was determined according to metabolic reactions listed in Additional file 1: Table S1.…”

Section: Methodology Of Mfamentioning

confidence: 99%

“…Glutamine and asparagine are the two most consumed amino acids in mammalian cell culture. To achieve high-specific growth rate, cell density, and productivity, the culture should be fed with high levels of glutamine or asparagine (Wlaschin and Hu 2006;Jain and Kumar 2008;Lu et al 2013;Sengupta et al 2011). Specifically, glutamine and asparagine are mutually related to each other not only in transportation but also in metabolism (Wipf et al 2002;Huang et al 2007;Kobayashi et al 2015;Xu et al 2014).…”

Section: Introductionmentioning

confidence: 99%

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Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Zhang

Wang

et al. 2016

Bioresour. Bioprocess.

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Background: Optimization of chemically defined medium has been a critical way to produce monoclonal antibody. Usually, amount of glutamine was added into the feed medium, but a half of asparagine was added. Our study found that asparagine was important in the antibody production phase. Increasing the ratio of asparagine to glutamine in feed medium for enhancement of antibody production in CHO-DHFR cell culture would be an efficient way. Results:We optimized the total amount and the ratio of the two vital amino acids in feed medium to increase antibody production. In this work, we have demonstrated that feeding medium of high ratio between asparagine and glutamine (FB-H) can enhance the cell density after reaching the stationary phase. Moreover, FB-H was shown to improve cell maintenance, and increased the antibody production. The metabolic flux analysis proved that ratio of asparagine to glutamine had little influence on glycolysis. Furthermore, the TCA cycle of FB-H was enhanced by 20 % compared to that of low ratio of asparagine to glutamine (FB-L). And the energy metabolism of FB-H was 22.6 % higher than that of FB-L. For the later, lactate can be less produced in FB-H. Conclusions:We should improve the ratio between asparagine and glutamine in feed medium properly under the premise of no influence on cell growth to achieve high mAb producing goal.

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Section: Methodology Of Mfamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Zhang

Wang

et al. 2016

Bioresour. Bioprocess.

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“…Screening of basal medium composition, nutrient additives in the feed medium for fed batch culture and physical parameters like pH and temperature, including assessment of temperature shift to mild hypothermia which is subsequent to initial growth phase, identifies the best set of design parameters for that clone to improve volumetric productivity. Such medium and process development efforts are important and have also been reviewed 74,75 . In the recent past, guidance from US-FDA has resulted in a push towards better characterization of processes to identify operating parameter space within which the performance of the process is robust in terms of productivity and product quality 76 .…”

Section: Upstream Process Developmentmentioning

confidence: 99%

Cell Culture Processes for Biopharmaceutical Manufacturing

Gadgil

2017

Current Science

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Recombinant proteins manufactured using animal cell culture processes comprise a significant fraction of biopharmaceuticals. With the expiry of patents on this class of therapeutics, there is also a significant interest in manufacture of biosimilar versions of such therapeutics. This article provides a birds-eye view of upstream process development for animal cell culture processes, with a focus on advances pertinent to the development of processes for biosimilars.

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“…The feed medium was prepared in a manner similar to that described by Wlaschin and Hu (2006). The feed rates of amino acids and phosphorus were determined by calculating the uptake rates of previous batch processes.…”

Section: Experimental Design and Statistical Analysismentioning

confidence: 99%

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

et al. 2012

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A serum-free medium (CHO-SFM) together with a fed-batch process was developed for the cultivation of a recombinant GS-CHO cell line producing TNFR-Fc. According to the metabolic characteristics of GS-CHO cell, a basal medium was prepared by supplementing DMEM:F12:RPMI1640 (2:1:1) with amino acids, insulin, transferrin, Pluronic F68 and some other ingredients. Statistical optimization approaches based on Plackett-Burman and central composite designs were then adopted to identify additional positive determinants and determine their optimal concentrations, which resulted in the final CHO-SFM medium formulations. The maximum antibody titer reached was 90.95 mg/l in the developed CHO-SFM, which was a 18 % and 10 fold higher than that observed in the commercial EX-CELL TM 302 medium (76.95 mg/l) and basal medium (8.28 mg/l), respectively. Subsequently, a reliable, reproducible and robust fed-batch strategy was designed according to the offline measurement of glucose, giving a final antibody yield of 378 mg/l, which was a threefold improvement over that in conventional batch culture (122 mg/l) using CHO-SFM. In conclusion, the use of design of experiment (DoE) method facilitated the development of CHO-SFM medium and fed-batch process for the production of recombinant antibody using GS-CHO cells.

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Fedbatch Culture and Dynamic Nutrient Feeding

Cited by 67 publications

References 70 publications

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Cell Culture Processes for Biopharmaceutical Manufacturing

Rational development of a serum-free medium and fed-batch process for a GS-CHO cell line expressing recombinant antibody

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