Zinc supplementation increases protein titer of recombinant CHO cells

Roca, Berta Capella; Miguez, Antonio Alarcon; Keenan, Joanne; Suda, Srinivas; Barron, Niall; O'Gorman, Donal J.; Doolan, Padraig; Clynes, Martin

doi:10.1007/s10616-019-00334-1

Cited by 9 publications

(8 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although the addition of copper to FAC B iron source to match the same copper concentration as present within FAC A improved the cell performance significantly, there was still a difference observed in cell performance compared to FAC A that may be due to further impurities. Other trace elements that have been either reported in literature for their positive effect on cell growth, viability or titer were for instance zinc and vanadium, 16,17 even though an excess of vanadium was also reported to cause a cytotoxicity within CHO cells 18,19 . In order to investigate whether impurities present within FAC A might have led to a toxic effect and thus might have caused a faster decline in cell culture viability compared to FAC B supplemented with copper, an in‐house synthesized low impurity FAC iron source (FAC Synt ; impurity profile presented in Table 1) was used and compared to the previously used FAC B + Cu 2+ condition.…”

Section: Resultsmentioning

confidence: 99%

Copper impurity of iron raw material contributes to improved cell culture performance

Weiss

Caspari

Merkel

et al. 2022

Biotechnology Progress

View full text Add to dashboard Cite

Cell culture medium (CCM) formulations are chemically defined to reduce lot‐to‐lot variability and complexity of the medium while still providing all essential nutrients supporting cell growth and productivity of various cell lines. However, raw material impurities may still introduce variations and inconsistencies to final CCM formulations. In one of our previous studies (Weiss et al. Biotechnol Prog. 2021;37(4):e3148), we have demonstrated the impact of iron raw material impurity on Chinese hamster ovary (CHO) cell performance and critical quality attributes (CQAs) of recombinant proteins within the Cellvento® 4CHO CCM platform by identifying manganese impurity as the main root cause for improved cell performance and altered glycosylation profiles. This study sought to investigate the impact of iron raw material impurities within another medium platform, namely EX‐CELL® Advanced CHO Fed‐Batch‐Medium. As opposed to previously published results, in this platform, copper instead of manganese impurity present within the used ferric ammonium citrate (FAC) iron source was responsible for an improved cell performance of a CHOZN® cell line and a slight difference in CQAs of the produced recombinant protein. The use of tightly controlled raw material specifications or the use of low impurity iron sources is therefore crucial to minimize the impact of impurities on cell performance in any CCM platform and thereby guarantee consistent and reproducible cell culture processes.

show abstract

Section: Resultsmentioning

confidence: 99%

Copper impurity of iron raw material contributes to improved cell culture performance

Weiss

Caspari

Merkel

et al. 2022

Biotechnology Progress

View full text Add to dashboard Cite

show abstract

“…During the development of a serum-free medium for CHO cells (SFM-F12) (25), reduction of putrescine supplementation to less than 200 g/liter was observed to have a dramatic negative effect on cell growth, resulting in a consistent drop on viability to less than 55% at concentrations lower than 100 g/liter. In SFM-F12 with increased putrescine supplementation up to 200 g/liter, a substantial negative effect in viable cell density (VCD) and viabilities was still observed (Fig.…”

Section: Cho-k1 Cells Require Supplementation Of Putrescine or L-ornithine For Healthy Growthmentioning

confidence: 99%

“…The latter was further supplemented with HEPES, linoleic acid (L1376), and glucose to mimic DMEM-F12 formulation. Both basal media were further supplemented with sodium selenite (S5261), recombinant insulin (I9279), ethanolamine (E0135), ammonium iron (III) citrate (F5879), polyvinyl alcohol, L-glutamine (Gibco, 25030024), nonessential amino acids (Gibco, 11140035), and putrescine dihydrochloride (P7505) (25). When indicated, 100 M L-ornithine or 1 mg/liter putrescine was added to each medium.…”

Section: Media Developmentmentioning

confidence: 99%

An arginase-based system for selection of transfected CHO cells without the use of toxic chemicals

Roca

Lao

Barron

et al. 2019

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

“…Efforts to further enhance mAb productivity and elicit desirable product quality profiles involve supplying trace metals to culture media in excess beyond basal provisions. For example, enhanced zinc supplementation has shown to increase mAb productivity, 2,5 galactosylation, 3 and suppress apoptosis 6 and combat oxidative stress 7 to prolong CHO culture. Copper supplementation can enhance the mAb productivity and induce lactate consumption and minimize waste metabolites 4 .…”

Section: Introductionmentioning

confidence: 99%

Effect of iron addition on mAb productivity and oxidative stress in Chinese hamster ovary culture

Graham

Mohammad

Liang

et al. 2021

Biotechnology Progress

View full text Add to dashboard Cite

Trace metals play a critical role in the development of culture media used for the production of therapeutic proteins. Iron has been shown to enhance the productivity of monoclonal antibodies during Chinese hamster ovary (CHO) cell culture. However, the redox activity and pro‐oxidant behavior of iron may also contribute toward the production of reactive oxygen species (ROS). In this work, we aim to clarify the influence of trace iron by examining the relationship between iron supplementation to culture media, mAb productivity and glycosylation, and oxidative stress interplay within the cell. Specifically, we assessed the impacts of iron supplementation on (a) mAb production and glycosylation; (b) mitochondria‐generated free hydroxyl radicals (ROS); (c) the cells ability to store energy during oxidative phosphorylation; and (d) mitochondrial iron concentration. Upon the increase of iron at inoculation, CHO cells maintained a capacity to rebound from iron‐induced viability lapses during exponential growth phase and improved mAb productivity and increased mAb galactosylation. Fluorescent labeling of the mitochondrial hydroxyl radical showed enhanced environments of oxidative stress upon iron supplementation. Additional labeling of active mitochondria indicated that, despite the enhanced production of ROS in the mitochondria, mitochondrial membrane potential was minimally impacted. By replicating iron treatments during seed train passaging, the CHO cells were observed to adapt to the shock of iron supplementation prior to inoculation. Results from these experiments demonstrate that CHO cells have the capacity to adapt to enhanced environments of oxidative stress and improve mAb productivity and mAb galactosylation with minimal perturbations to cell culture.

show abstract

Zinc supplementation increases protein titer of recombinant CHO cells

Cited by 9 publications

References 28 publications

Copper impurity of iron raw material contributes to improved cell culture performance

Copper impurity of iron raw material contributes to improved cell culture performance

An arginase-based system for selection of transfected CHO cells without the use of toxic chemicals

Effect of iron addition on mAb productivity and oxidative stress in Chinese hamster ovary culture

Contact Info

Product

Resources

About