Advantages of AlaGln as an additive to cell culture medium: use with anti-CD20 chimeric antibody-producing POTELLIGENT™ CHO cell lines

Imamoto, Y.; Tanaka, Hiromu; Takahashi, Ken; Konno, Yoshinobu; Suzawa, Toshiyuki

doi:10.1007/s10616-012-9468-8

Cited by 21 publications

(12 citation statements)

References 21 publications

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“…The analysis of cell-specific mAb production reveals that only AQ outperformed, with a maximum cell-specific mAb production rate 34 % higher than the control (revealing a borderline statistical relevance). Nevertheless, the finding matches the results obtained by Imamoto et al ( 2013 ) (Fig. 4 a).…”

Section: Resultssupporting

confidence: 93%

“…Due to their diversity and their different boosting properties, dipeptides emerge as feasible defined medium components able to improve cell productivities. Dipeptides such as l -alanyl- l -glutamine (AQ) supplanted the use of heat-sensitive glutamine (Minamoto et al 1991 ; Atanassov et al 1998 ; Imamoto et al 2010 ; Kim et al 2012a , b ; Imamoto et al 2013 ). Glycyl- l -glutamine (GQ) additions were studied, revealing beneficial effects in murine hybridoma cell cultures (Christie and Butler 1994 ).…”

Section: Introductionmentioning

confidence: 99%

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Tracking dipeptides at work-uptake and intracellular fate in CHO culture

et al. 2016

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Market demands for monoclonal antibodies (mAbs) are steadily increasing worldwide. As a result, production processes using Chinese hamster ovary cells (CHO) are in the focus of ongoing intensification studies for maximizing cell-specific and volumetric productivities. This includes the optimization of animal-derived component free (ADCF) cultivation media as part of good cell culture practice. Dipeptides are known to improve CHO culture performance. However, little or even conflicting assumptions exist about their putative import and functionality inside the cells. A set of well-known performance boosters and new dipeptide prospects was evaluated. The present study revealed that dipeptides are indeed imported in the cells, where they are decomposed to the amino acids building blocks. Subsequently, they are metabolized or, unexpectedly, secreted to the medium. Monoclonal antibody production boosting additives like l-alanine-l-glutamine (AQ) or glycyl-l-glutamine (GQ) can be assigned to fast or slow dipeptide uptake, respectively, thus pinpointing to the need to study dipeptide kinetics and to adjust their feeding individually for optimizing mAb production.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-016-0221-0) contains supplementary material, which is available to authorized users.

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Section: Resultssupporting

confidence: 93%

Section: Introductionmentioning

confidence: 99%

Tracking dipeptides at work-uptake and intracellular fate in CHO culture

et al. 2016

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“…Prevention of cysteine degradation may assist modulation of the cell culture redox environment, thus resulting in higher q p . Application of tyrosine‐, histidine‐, or glutamine‐containing dipeptides, substitution of glutamine with alpha‐ketoglutarate (alpha‐KG), or altering the glutamine levels in media have been reported to lower lactate and ammonia production, thus providing better pH maintenance, improved q p , and overall process yield . Yeast extracts produced a beneficial effect on q p by maintaining the mammalian target of rapamycin pathway activity and enhancing protein translation .…”

Section: Introductionmentioning

confidence: 99%

Increased MSX level improves biological productivity and production stability in multiple recombinant GS CHO cell lines

Tian

Oliveira

et al. 2020

Engineering in Life Sciences

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Increasing cell culture productivity of recombinant proteins via process improvements is the primary focus for research groups within biologics manufacturing. Any recommendations to improve a manufacturing process obviously must be effective, but also be robust, scalable, and with product quality comparable to the original process. In this study, we report that three different GS−/− CHO cell lines developed in media containing a standard concentration of the selection agent methionine sulfoximine (MSX), but then exposed to increased MSX concentrations during seed train expansion, achieved titer increases of 10–19%. This result was observed in processes already considerably optimized. Expanding the cells with a higher MSX concentration improved cell line production stability with increased culture age. Production cultures in 500‐L and 1000‐L bioreactors replicated laboratory results using 5‐L bioreactors, demonstrating process robustness and scalability. Furthermore, product quality attributes of the final drug substance using the higher MSX process were comparable with those from cells expanded in media with the standard selection MSX concentration. Subsequent mechanistic investigations confirmed that the cells were not altered at the genetic level in terms of integration profiles or gene copy number, nor transcriptional levels of glutamine synthetase, heavy chain, or light chain genes. This study provides an effective and applicable strategy to improve the productivity of therapeutic proteins for biologics manufacturing.

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“…Stressful conditions like hyperosmotic stress, nutrient limitation and ammonia concentration have been correlated to increased levels of apoptotic cells [35–37]. However, neither cell line shows increased levels of apoptosis (Figure 4A,B).…”

Section: Resultsmentioning

confidence: 99%

Investigation of cell line specific responses to pH inhomogeneity and consequences for process design

Paul

Hartmann

Posch³

et al. 2020

Engineering in Life Sciences

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With increasing bioreactor volumes, the mixing time of the reactor increases as well, which creates an inhomogeneous environment for the cells. This can result in impaired process performance in large‐scale production reactors. Particularly the addition of base through the reactor headspace can be problematic, since it creates an area, where cells are repeatedly exposed to an increased pH. The aim of this study is to simulate this large‐scale phenomenon at lab‐scale and investigate its impact. Two different cell lines were exposed to pH amplitudes of a maximal magnitude of 0.05 units (pH of 6.95). Both cell lines showed similar responses, like decreased viable cell counts, but unaffected lactate levels. However, cell line B showed an initially increased specific productivity in response to the introduced amplitudes, whereas cell line A showed a consistently lower specific productivity. Furthermore, the time point at which base addition is started influences the impact, which pH amplitudes have on process performance. When pH control was started earlier in the process, maximal viable cell counts decreased and the lactate metabolic shift was less pronounced. These results show that the potential negative impact of pH amplitudes can be minimized by strategic process design.

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Advantages of AlaGln as an additive to cell culture medium: use with anti-CD20 chimeric antibody-producing POTELLIGENT™ CHO cell lines

Cited by 21 publications

References 21 publications

Tracking dipeptides at work-uptake and intracellular fate in CHO culture

Tracking dipeptides at work-uptake and intracellular fate in CHO culture

Increased MSX level improves biological productivity and production stability in multiple recombinant GS CHO cell lines

Investigation of cell line specific responses to pH inhomogeneity and consequences for process design

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