Optimization of chemically defined feed media for monoclonal antibody production in Chinese hamster ovary cells

Kishishita, Shohei; Katayama, Satoshi; Kodaira, Kunihiko; Takagi, Yoshinori; Matsuda, Hiroki; Okamoto, Hiroshi; Takuma, Shinya; Hirashima, Chikashi; Aoyagi, Haruhiko

doi:10.1016/j.jbiosc.2014.11.022

Cited by 44 publications

(31 citation statements)

References 58 publications

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“…SLC7A11, responsible for Cys import, is of specific interest as only inhibition of SLC7A11 by SAS induced a rapid loss of cell viability in all cell populations, consistent with findings that SLC7A11 activity is strongly associated with GSH synthesis and SAS‐mediated inhibition of cystine uptake via SLC7A11 induces apoptotic cell death (Chung et al, ; Gout et al, ; Shih et al, ; Timmerman et al, ). Cystine transport might therefore be a competitive bottleneck to production of cell and MAb product biomass that may itself induce stress responses (Lee et al, ) which may be amenable to either genetic cell engineering or improved media design such as supplementing cultures with cysteine containing dipeptides or tripeptides that have a higher solubility as reported by others (Kang, Mullen, Miranda, & Deshpande, ; Kishishita et al, ). However, evidence would suggest that Cys levels in mammalian cells are tightly regulated as excess Cys is cytotoxic (Stipanuk, Dominy, Lee, & Coloso, ).…”

Section: Resultsmentioning

confidence: 99%

Control of amino acid transport into Chinese hamster ovary cells

Geoghegan

Arnall

Hatton³

et al. 2018

Biotech & Bioengineering

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Amino acid transporters (AATs) represent a key interface between the cell and its environment, critical for all cellular processes: Energy generation, redox control, and synthesis of cell and product biomass. However, very little is known about the activity of different functional classes of AATs in Chinese hamster ovary (CHO) cells, how they support cell growth and productivity, and the potential for engineering their activity and/or the composition of amino acids in growth media to improve CHO cell performance in vitro. In this study, we have comparatively characterized AAT expression in untransfected and monoclonal antibody (MAb)‐producing CHO cells using transcriptome analysis by RNA‐seq, and mechanistically dissected AAT function using a variety of transporter‐specific chemical inhibitors, comparing their effect on cell proliferation, recombinant protein production, and amino acid transport. Of a possible 56 mammalian plasma membrane AATs, 16 AAT messenger RNAs (mRNAs) were relatively abundant across all CHO cell populations. Of these, a subset of nine AAT mRNAs were more abundant in CHO cells engineered to produce a recombinant MAb. Together, upregulated AATs provide additional supply of specific amino acids overrepresented in MAb biomass compared to CHO host cell biomass, enable transport of synthetic substrates for glutathione synthesis, facilitate transport of essential amino acids to maintain active protein synthesis, and provide amino acid substrates for coordinated antiport systems to maintain supplies of proteinogenic and essential amino acids.

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Section: Resultsmentioning

confidence: 99%

Control of amino acid transport into Chinese hamster ovary cells

Geoghegan

Arnall

Hatton³

et al. 2018

Biotech & Bioengineering

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“…To solve this the amino acids could also be added as di- or tripeptides. It was shown that amino acids supplemented in the form of a dipeptide or tripeptide improved their solubility, yet resulted in similar culture effects (Kishishita et al 2015). …”

Section: Discussionmentioning

confidence: 99%

Selection of chemically defined media for CHO cell fed-batch culture processes

et al. 2016

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Two CHO cell clones derived from the same parental CHOBC® cell line and producing the same monoclonal antibody (BC-G, a low producing clone; BC-P, a high producing clone) were tested in four basal media in all possible combinations with three feeds (=12 conditions) in fed-batch cultures. Higher amino acid feeding did not always lead to higher mAb production. The two clones showed differences in cell physiology, metabolism and optimal medium-feed combinations. During the phase transitions of all cultures, cell metabolism showed a shift represented by lower specific consumption and production rates, except for the specific glucose consumption rate in cultures fed by Actifeed A/B. The BC-P clone fed by Actifeed A/B showed a threefold cell volume increase and an increase of the specific consumption rate of glucose in the stationary phase. Since feeding was based on glucose this resulted in accumulation of amino acids for this feed, while this did not occur for the poorer feed (EFA/B). The same feed also led to an increase of cell size for the BC-G clone, but to a lesser extent.Electronic supplementary materialThe online version of this article (doi:10.1007/s10616-016-0036-5) contains supplementary material, which is available to authorized users.

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“…The extent of electronic and hydrophobic interaction could vary depending on the feed stream matrix properties, antibody properties, and depth filter characteristics. Further, the recent trend of increasing antibody titers using enriched growth media and high density culture (Huang et al, 2010;Kishishita et al, 2015;Templeton, Dean, Reddy, & Young, 2013;Xie & Wang, 1996) results in higher level of product-related species in the bioreactor, and thus causes additional challenges on subsequent purification steps following clarification. Among others, impurities, such as HMW and LMW are very critical for the safety and efficacy of the protein in clinical use (Schenauer, Flynn, & Goetze, 2013).…”

mentioning

confidence: 99%

Control of antibody high and low molecular weight species by depth filtration‐based cell culture harvesting

Mayani

Song

et al. 2019

Biotech & Bioengineering

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Depth filtration-based harvesting is widely used in mAb manufacturing to remove cell and process-related impurities. However, it has not been studied on control of product-related impurities, which are very critical for product quality. In this article, we studied the interactions of depth filter with high and low molecular weight species (HMWs and LMWs) for their direct removal from cell culture. The process parameters (filter, loading, temperature, and flux) were evaluated for adsorption of HMWs and LMWs by depth filters. The adsorption is significantly dependent on filter media and loading capacity and is mainly on the basis of hydrophobic interaction during harvesting. The HMW and LMW species were characterized as HMW1, HMW2, LMW1, and LMW2. The increasing binding from LMW2 to LMW1, HMW1, and HMW2 is correlated with their increasing hydrophobicity score. Adsorption using enriched HMW sample demonstrated similar total protein binding capacity (36-40 g/m 2 ) between depth filters D0HC and X0HC. However, X0HC has stronger HMW binding than D0HC (71% vs 43% of bound protein), indicating more hydrophobic interaction in X0HC. HMW2 DBC on X0HC reached 12 g/m 2 , similar to protein binding on hydrophobic interaction membrane adsorbers. Further study showed LMW can induce HMW formation. This study provides a critical understanding of HMW and LMW interaction with depth filters. The strategy of HMW and LMW control by depth filtration-based harvesting was implemented successfully in mAb manufacturing. K E Y W O R D Scell culture harvesting, depth filtration, high molecular weight species, low molecular weight species, monoclonal antibody, protein adsorption

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Optimization of chemically defined feed media for monoclonal antibody production in Chinese hamster ovary cells

Cited by 44 publications

References 58 publications

Control of amino acid transport into Chinese hamster ovary cells

Control of amino acid transport into Chinese hamster ovary cells

Selection of chemically defined media for CHO cell fed-batch culture processes

Control of antibody high and low molecular weight species by depth filtration‐based cell culture harvesting

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