Investigation into the impact of tyrosine on the product formation and quality attributes of mAbs in rCHO cell cultures

Zhang, Weijian; Liu, Xuping; Tang, Hongping; Zhang, Xinran; Zhou, Yanan; Fan, Li; Hai-bin, Wang; Tan, Wen‐Song; Zhao, Liang

doi:10.1007/s00253-020-10744-3

Cited by 10 publications

(5 citation statements)

References 63 publications

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“…Apart from feed medium containing tyrosine, 88 mM tyrosine solution was added in the high concentration tyrosine group. High concentration tyrosine promoted the cell growth and reduced the osmolality of bioreactor culture environment and increased the production of mAbs (Zhang et al 2020a ).…”

Section: Amino Acidmentioning

confidence: 99%

Progress in fed-batch culture for recombinant protein production in CHO cells

Lin

et al. 2023

Appl Microbiol Biotechnol

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Nearly 80% of the approved human therapeutic antibodies are produced by Chinese Hamster Ovary (CHO) cells. To achieve better cell growth and high-yield recombinant protein, fed-batch culture is typically used for recombinant protein production in CHO cells. According to the demand of nutrients consumption, feed medium containing multiple components in cell culture can affect the characteristics of cell growth and improve the yield and quality of recombinant protein. Fed-batch optimization should have a connection with comprehensive factors such as culture environmental parameters, feed composition, and feeding strategy. At present, process intensification (PI) is explored to maintain production flexible and meet forthcoming demands of biotherapeutics process. Here, CHO cell culture, feed composition in fed-batch culture, fed-batch culture environmental parameters, feeding strategies, metabolic byproducts in fed-batch culture, chemostat cultivation, and the intensified fed-batch are reviewed. Key points • Fed-batch culture in CHO cells is reviewed. • Fed-batch has become a common technology for recombinant protein production. • Fed batch culture promotes recombinant protein production in CHO cells.

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Section: Amino Acidmentioning

confidence: 99%

Progress in fed-batch culture for recombinant protein production in CHO cells

Lin

et al. 2023

Appl Microbiol Biotechnol

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“…Furthermore, while fed-batch processes persist as the forefront approach in biopharmaceutical manufacturing, perfusion or continuous processes are being increasingly favored to elevate productivity (Yongky et al 2019 ; Xu et al 2020 ). These intensified biopharma processes require highly concentrated feeds or media for nutrient replenishment, serving to prevent any reduction in productivity due to dilution effects (Zhang et al 2020 ). However, formulating such concentrated feeds or media is challenging owing to the instability (e.g., glutamine) and limited solubility of specific amino acids (e.g., cystine and tyrosine) as well as other critical components (e.g., keto acids) (Christie and Butler 1994 ; Tabata and Hashimoto 2007 ; Kang et al 2012 ; Nishino et al 2022 ).…”

Section: Introductionmentioning

confidence: 99%

“…This aromatic ring in L-Tyr contains a large number of hydrophobic carbon atoms that offset the hydrogen bonding interactions formed by the hydroxyl group with water molecules, resulting in limited solubility in aqueous solutions. L-Tyr plays a crucial role as an indispensable amino acid that regulates cellular metabolism, protein synthesis, and the overall quality of products in CHO cell culture (Yu et al 2011 ; Feeney et al 2013 ; Zhang et al 2020 ). In traditional industrialized biomanufacturing, a separate L-Tyr stock solution, adjusted to an extreme alkaline pH to elevate it away from its isoelectric point (pI = 5.64), has been used in addition to the primary pH-neutral feed medium (Lee et al 2013 ; Zimmer et al 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Exploring metabolic effects of dipeptide feed media on CHO cell cultures by in silico model-guided flux analysis

Park,

Song,

Choi

et al. 2024

Appl Microbiol Biotechnol

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There is a growing interest in perfusion or continuous processes to achieve higher productivity of biopharmaceuticals in mammalian cell culture, specifically Chinese hamster ovary (CHO) cells, towards advanced biomanufacturing. These intensified bioprocesses highly require concentrated feed media in order to counteract their dilution effects. However, designing such condensed media formulation poses several challenges, particularly regarding the stability and solubility of specific amino acids. To address the difficulty and complexity in relevant media development, the biopharmaceutical industry has recently suggested forming dipeptides by combining one from problematic amino acids with selected pairs to compensate for limitations. In this study, we combined one of the lead amino acids, L-tyrosine, which is known for its poor solubility in water due to its aromatic ring and hydroxyl group, with glycine as the partner, thus forming glycyl-L-tyrosine (GY) dipeptide. Subsequently, we investigated the utilization of GY dipeptide during fed-batch cultures of IgG-producing CHO cells, by changing its concentrations (0.125 × , 0.25 × , 0.5 × , 1.0 × , and 2.0 ×). Multivariate statistical analysis of culture profiles was then conducted to identify and correlate the most significant nutrients with the production, followed by in silico model-guided analysis to systematically evaluate their effects on the culture performance, and elucidate metabolic states and cellular behaviors. As such, it allowed us to explain how the cells can more efficiently utilize GY dipeptide with respect to the balance of cofactor regeneration and energy distribution for the required biomass and protein synthesis. For example, our analysis results uncovered specific amino acids (Asn and Gln) and the 0.5 × GY dipeptide in the feed medium synergistically alleviated the metabolic bottleneck, resulting in enhanced IgG titer and productivity. In the validation experiments, we tested and observed that lower levels of Asn and Gln led to decreased secretion of toxic metabolites, enhanced longevity, and elevated specific cell growth and titer. Key points • Explored the optimal Tyr dipeptide for the enhanced CHO cell culture performance • Systematically analyzed effects of dipeptide media by model-guided approach • Uncovered synergistic metabolic utilization of amino acids with dipeptide Graphical abstract

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“…For instance, glutamine limitation reduced cell growth but did not affect specific productivity, while asparagine limitation did not significantly affect either of these parameters. Cysteine and tyrosine limitations have been reported to be detrimental to growth and productivity 12,13 …”

Section: Introductionmentioning

confidence: 99%

“…Cysteine and tyrosine limitations have been reported to be detrimental to growth and productivity. 12,13 Varying single amino acid concentrations may; however, not reflect the physiological effects observed in the context of the availability of other amino acids. For example, competition for uptake between cystine and glutamate is known to inhibit their uptake upon the addition of the other amino acid.…”

mentioning

confidence: 99%

Amino acid abundance and composition in cell culture medium affects trace metal tolerance and cholesterol synthesis

Rawat

Bhambri

Pandey

et al. 2022

Biotechnology Progress

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Amino acid compositions of cell culture media are empirically designed to enhance cell growth and productivity and vary both across media formulations and over the course of culture due to imbalance in supply and consumption. The interconnected nature of the amino acid transporters and metabolism suggests that changes in amino acid composition can affect cell physiology. In this study, we explore the effect of a step change in amino acid composition from a DMEM: F12-based medium to a formulation varying in relative abundances of all amino acids, evaluated at two amino acid concentrations (lean LAA vs. rich HAA). Cell growth was inhibited in LAA but not HAA. In addition to the expected effects on expression of the cell cycle, amino acid response and mTOR pathway genes in LAA, we observed an unanticipated effect on zinc uptake and efflux genes. This was accompanied by a lower tolerance to zinc supplementation in LAA but not in the other formulations. Histidine was sufficient but not necessary to prevent such zinc toxicity. Additionally, an unanticipated downregulation of genes in the cholesterol synthesis pathway was observed in HAA, accompanied by an increase in cellular cholesterol content, which may depend on the relative abundances of glutamine and other amino acids. This study shows that changes in the amino acid composition without any evident effect on growth may have profound effects on metabolism. Such analyses can help rationalize the designing of medium and feed formulations for bioprocess applications beyond replenishment of consumed components.

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Investigation into the impact of tyrosine on the product formation and quality attributes of mAbs in rCHO cell cultures

Cited by 10 publications

References 63 publications

Progress in fed-batch culture for recombinant protein production in CHO cells

Progress in fed-batch culture for recombinant protein production in CHO cells

Exploring metabolic effects of dipeptide feed media on CHO cell cultures by in silico model-guided flux analysis

Amino acid abundance and composition in cell culture medium affects trace metal tolerance and cholesterol synthesis

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