Effect of Ca2+ and Mg2+ concentration in culture medium on the activation of recombinant factor IX produced in Chinese hamster ovary cells

Kim, Wonhee; Kim, Jung-Seop; Yoon, Yeup; Lee, Gyun Min

doi:10.1016/j.jbiotec.2009.06.001

Cited by 12 publications

(7 citation statements)

References 22 publications

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“…When EDTA was added on D0 (at the inoculation stage) the peak viable cell density (VCD) remained close to the seeding density, indicating a hampering of cell growth. This result is consistent with the well-known fact that EDTA is toxic to cells [35]. The decline in the viable cell densities due to EDTA addition during the growth phase can also be attributed to the removal of such trace metals as Ca ++ , Zn ++ etc.…”

Section: Discussionsupporting

confidence: 90%

Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

2017

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Abstract:In order to meet desired drug product quality targets, the glycosylation profile of biotherapeutics such as monoclonal antibodies (mAbs) must be maintained consistently during manufacturing. Achieving consistent glycan distribution profiles requires identifying factors that influence glycosylation, and manipulating them appropriately via well-designed control strategies. Now, the cell culture media supplement, MnCl 2 , is known to alter the glycosylation profile in mAbs generally, but its effect, particularly when introduced at different stages during cell growth, has yet to be investigated and quantified. In this study, we evaluate the effect of time-dependent addition of MnCl 2 on the glycan profile quantitatively, using factorial design experiments. Our results show that MnCl 2 addition during the lag and exponential phases affects the glycan profile significantly more than stationary phase supplementation does. Also, using a novel computational technique, we identify various combinations of glycan species that are affected by this dynamic media supplementation scheme, and quantify the effects mathematically. Our experiments demonstrate the importance of taking into consideration the time of addition of these trace supplements, not just their concentrations, and our computational analysis provides insight into what supplements to add, when, and how much, in order to induce desired changes.

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Section: Discussionsupporting

confidence: 90%

Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

2017

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“…49 Kim and colleagues studied the effect of Ca 2+ and Mg 2+ concentration in serum-free culture on the activation of recombinant factor IX (aFIX) produced in CHO cells. 53 Ca 2+ at 0.5 mM was found to inhibit the conversion of aFIX to the undesirable activated factor IX (FIXa) at the end stage of cell culture when compared to that of cell culture containing 1.0 mM Ca 2+ . The various effects of these media supplements in different cell culture denote the importance of using well-defined media for the production of recombinant therapeutics.…”

Section: Cho Process Engineering Strategiesmentioning

confidence: 99%

“…Adenosine causes growth arrest in CHO cell culture and leads to an increase in intracellular ATP in mammalian cells, thereby enhancing protein expression capacity . Kim and colleagues studied the effect of Ca 2+ and Mg 2+ concentration in serum-free culture on the activation of recombinant factor IX (aFIX) produced in CHO cells . Ca 2+ at 0.5 mM was found to inhibit the conversion of aFIX to the undesirable activated factor IX (FIXa) at the end stage of cell culture when compared to that of cell culture containing 1.0 mM Ca 2+ .…”

Section: Cho Engineering Strategiesmentioning

confidence: 99%

Application of Multi-Omics Techniques for Bioprocess Design and Optimization in Chinese Hamster Ovary Cells

et al. 2014

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Significant improvements in the productivity and quality of therapeutic proteins produced in Chinese hamster ovary (CHO) cells have been reported since their establishment as host cells for biopharmaceutical production. Initial advances in the field focused on engineering strategies to manipulate genes associated with proliferation, apoptosis, and various metabolic pathways. Process engineering efforts to optimize culture media, batch-feeding strategies and culture conditions, including temperature and osmolarity, were also reported. More recently, focus has shifted toward enhancing process consistency and product quality using systems biology quality by design-based approaches during process development. Integration of different data generated using omics technologies, such as genomics, transcriptomics, proteomics and metabolomics, has facilitated a greater understanding of CHO cell biology. These techniques have enabled the provision of global information on dynamic changes in cellular components associated with different phenotypes. Using systems biology to understand these important host cells at the cellular level will undoubtedly result in further progression in the development and expression of biopharmaceutical products in CHO cells.

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“…It may be increased by 30-50% upon addition of 1 nM of methyl testosterone to the culture medium [38] or doubled after the addition of phorbol 12-myristate, 13-acetate, and calcium ionophore [39]. The appearance of undesired activated FIX in the culture medium can be controlled by decreasing the Ca 2+ ions concentration to 0.5 mM from 1.12 mM [40]; in another study of the same group, it was found that an increase of Ca 2+ ions to 1.3 mM leads to a 30% increase in the production of FIX, without a significant rise in the FIXa level [41]. …”

Section: Improvements In Recombinant Fix Productionmentioning

confidence: 99%

Coagulation Factor IX for Hemophilia B Therapy

et al. 2012

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Factor IX is a zymogen enzyme of the blood coagulation cascade. Inherited absence or deficit of the IX functional factor causes bleeding disorder hemophilia B, which requires constant protein replacement therapy. Reviewed herein are the current state in the manufacturing of FIX, improved variants of the recombinant protein for therapy, transgenic organisms for obtaining FIX, and the advances in the gene therapy of hemophilia B.

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Effect of Ca2+ and Mg2+ concentration in culture medium on the activation of recombinant factor IX produced in Chinese hamster ovary cells

Cited by 12 publications

References 22 publications

Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

Controlling the Glycosylation Profile in mAbs Using Time-Dependent Media Supplementation

Application of Multi-Omics Techniques for Bioprocess Design and Optimization in Chinese Hamster Ovary Cells

Coagulation Factor IX for Hemophilia B Therapy

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