Enhanced productivity of Protein C by recombinant human cells in automated fed-batch cultures

Kyung, Yun Seung; Hu, Wei Shou

doi:10.1007/bf00749398

Cited by 4 publications

(3 citation statements)

References 7 publications

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“…Enhancements in cell concentration and culture longevity have also been achieved through use of fed-batch mode of operation using strategies ranging from addition of medium concentrates (Bibila et al, 1994), through use of automated online nutrient feeding (Kyung and Hu, 1995;Zhou et al, 1995Zhou et al, , 1997, and through application of stoichiometric analysis of animal cell growth in medium/feeding design Wang, 1996, 1997).…”

Section: Introductionmentioning

confidence: 98%

Metabolism of PER.C6^TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

Maranga

Goochee

2006

Biotech & Bioengineering

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This is the first study to examine PER.C6 cell glucose/energy and glutamine metabolism with fed-batch cultures at controlled low glutamine, low glucose, and simultaneous low glucose and low glutamine levels. PER.C6(TM) cell metabolism was investigated in serum-free suspension bioreactors at two-liter scale. Control of glucose and/or glutamine concentrations had a significant effect on cellular metabolism leading to an increased efficiency of nutrient utilization, altered byproduct synthesis, while having no effect on cell growth rate. Cultivating cells at a controlled glutamine concentration of 0.25 mM reduced q(Gln) and q(NH(4)(+)) by approximately 30%, q(Ala) 85%, and q(NEAA) 50%. The fed-batch control of glutamine also reduced the overall accumulation of ammonium ion by approximately 50% by minimizing the spontaneous chemical degradation of glutamine. No major impact upon glucose/energy metabolism was observed. Cultivating cells at a glucose concentration of 0.5 mM reduced q(Glc) about 50% and eliminated lactate accumulation. Cells exhibited a fully oxidative metabolism with Y(O(2)/Glc) of approximately 6 mol/mol. However, despite no increase in q(Gln), an increased ammonium ion accumulation and Y(NH(4)(+)/Gln) were also observed. Effective control of lactate and ammonium ion accumulation by PER.C6 cells was achieved using fed-batch with simultaneously controlled glucose and glutamine. A fully oxidative glucose metabolism and a complete elimination of lactate production were obtained. The q(Gln) value was again reduced and, despite an increased q(NH(4)(+)) compared with batch culture, ammonium ion levels were typically lower than corresponding ones in batch cultures, and the accumulation of non-essential amino acids (NEAA) was reduced about 50%. In conclusion, this study shows that PER.C6 cell metabolism can be confined to a state with improved efficiencies of nutrient utilization by cultivating cells in fed-batch at millimolar controlled levels of glucose and glutamine. In addition, PER.C6 cells fall into a minority category of mammalian cell lines for which glutamine plays a minor role in energy metabolism.

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Section: Introductionmentioning

confidence: 98%

Metabolism of PER.C6^TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

Maranga

Goochee

2006

Biotech & Bioengineering

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“…This strategy has significantly increased cellular concentrations, the cell integral, and the recombinant protein production in CHO cells [65], hybridomas [66], myeloma [67], and 293 human kidney cells [68]. However, despite its effectiveness, the high cost, which may involve feeding all the nutrients of the medium and the high risk of increasing the osmolality of the medium, may be factors against the use of these feeds.…”

Section: Feeds and Hydrolysatesmentioning

confidence: 99%

5.3 Nutrient Media for Cell Culture Technology

Altamirano¹,

Berríos²,

Vergara³

2014

Animal Cell Biotechnology

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“…Higher productivities can be achieved by adjusting the culture conditions so that they can be maintained at the optimum conditions. Kyung and Hu (1995) reported enhanced productivity of protein C by recombinant human cells in automated fed-batch cultures. Productivities often depends on components of culture media including autocrine factors and some components which are derived from cells after their lyses.…”

Section: Introductionmentioning

confidence: 99%

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et al. 1997

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Animal cell perfusion high density culture is often adopted for the production of biologicals in industry. In high density culture sometimes the productivity of biologicals has been found to be enhanced. Especially in immobilized animal cell culture, significant increase in the productivity has been reported. We have found that the specific monoclonal antibody (MAb) productivity of an immobilized hybridoma cell is enhanced more than double. Several examples of enhancing productivities have been also shown by collagen immobilized cells. Immobilized cells involve some different points from non-immobilized cells in high density culture: In immobilized culture, some cells are contacted together, resulting in locally much higher cell concentration more than 10(8) cells/ml. Information originating from a cell can be easily transduced to the others in immobilized culture because the distance between cells is much nearer. Here we have performed collagen gel immobilized culture of recombinant BHK cells which produce a human IgG monoclonal antibody in a protein-free medium for more than three months. In this high density culture a stabilized monoclonal antibody production was found with around 8 times higher specific monoclonal antibody productivity compared with that in a batch serum containing culture. No higher MAb productivity was observed using a conditioned medium which was obtained from the high density culture, indicating that no components secreted from the immobilized cells work for enhancing monoclonal antibody production. The MAb productivity by the non-immobilized cells obtained by dissolving collagen using a collagenase gradually decreased and returned to the original level in the batch culture using a fresh medium. This suggests that the direct contact of the cells or a very close distance between the cells has something to do with the enhancement of the MAb productivity, and the higher productivity is kept for a while in each cell after they are drawn apart.

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Enhanced productivity of Protein C by recombinant human cells in automated fed-batch cultures

Cited by 4 publications

References 7 publications

Metabolism of PER.C6^TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

Metabolism of PER.C6^TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

5.3 Nutrient Media for Cell Culture Technology

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Enhanced productivity of Protein C by recombinant human cells in automated fed-batch cultures

Cited by 4 publications

References 7 publications

Metabolism of PER.C6TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

Metabolism of PER.C6TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

5.3 Nutrient Media for Cell Culture Technology

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Metabolism of PER.C6^TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels

Metabolism of PER.C6^TM cells cultivated under fed‐batch conditions at low glucose and glutamine levels