Amino acids in the cultivation of mammalian cells

Salazar, Andrew; Keusgen, Michael; Hagen, Jörg von

doi:10.1007/s00726-016-2181-8

Cited by 95 publications

(83 citation statements)

References 86 publications

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“…In CHO cell culture, amino acids are supplied in the growth medium and/or produced via biosynthetic pathways. These are required to support cellular functions, such as cell growth, and utilized as building blocks for protein synthesis . Through the catabolism of amino acids, the cell can utilize the carbon backbones for the formation of TCA cycle intermediates, which are to be used in the central metabolic pathways (Figure ).…”

Section: An Overview Of Cho Metabolismmentioning

confidence: 99%

“…These are required to support cellular functions, such as cell growth, and utilized as building blocks for protein synthesis. [44,45] Through the catabolism of amino acids, the cell can utilize the carbon backbones for the formation of TCA cycle intermediates, which are to be used in the central metabolic pathways (Figure 1). When these are supplied in excess, a wasteful cellular metabolism will lead to the formation of by-products, in particular ammonium.…”

Section: Amino Acid Metabolism In Cho Cell Culturementioning

confidence: 99%

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Impact of CHO Metabolism on Cell Growth and Protein Production: An Overview of Toxic and Inhibiting Metabolites and Nutrients

Pereira

Kildegaard

Andersen

2018

Biotechnology Journal

152

146

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For over three decades, Chinese hamster ovary (CHO) cells have been the chosen expression platform for the production of therapeutic proteins with complex post-translational modifications. However, the metabolism of these cells is far from perfect and optimized, and requires substantial know how and process optimization and monitoring to perform efficiently. One of the main reasons for this is the production and accumulation of toxic and growth-inhibiting metabolites during culture. Lactate and ammonium are the most known, but many more have been identified. In this review, an overview of metabolites that deplete and accumulate throughout the course of cultivations with toxic and growth inhibitory effects to the cells is presented. Further, an overview of the CHO metabolism with emphasis to metabolic pathways of amino acids, glutathione (GSH), and related compounds which have growth-inhibiting and/or toxic effect on the cells is provided. Additionally, relevant publications which describe the applications of metabolomics as a powerful tool for revealing which reactions occur in the cell under certain conditions are surveyed and growth-inhibiting and toxic metabolites are identified. Also, a number of resources that describe the cellular mechanisms of CHO and are available on-line are presented. Finally, the application of this knowledge for bioprocess and medium development and cell line engineering is discussed.

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Section: An Overview Of Cho Metabolismmentioning

confidence: 99%

Section: Amino Acid Metabolism In Cho Cell Culturementioning

confidence: 99%

Impact of CHO Metabolism on Cell Growth and Protein Production: An Overview of Toxic and Inhibiting Metabolites and Nutrients

Pereira

Kildegaard

Andersen

2018

Biotechnology Journal

152

146

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“…However, understanding the metabolism of individual nutrients could provide a more objective approach to experimental design and process optimization. Although amino acids are important nutrients, essential for many biological functions of cells and complex organisms, there are relatively few publications on the effects of individual amino acid levels on the performance of CHO cell cultures . In some cases, it has been noted that replacement of depleted amino acids has not led to process improvements .…”

Section: Introductionmentioning

confidence: 99%

Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed‐batch cultures

Ghaffari

Jardon

Krahn

et al. 2019

Biotechnology Progress

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Amino acid availability is a key factor that can be controlled to optimize the productivity of fed‐batch cultures. To study amino acid limitation effects, a serum‐free chemically defined basal medium was formulated to exclude the amino acids that became depleted in batch culture. The effect of limiting glutamine, asparagine, and cysteine on the cell growth, metabolism, antibody productivity, and product glycosylation was investigated in three Chinese hamster ovary (CHO) cell lines (CHO‐DXB11, CHO‐K1SV, and CHO‐S). Cysteine limitation was detrimental to both cell proliferation and productivity for all three CHO cell lines. Glutamine limitation reduced growth but not cell specific productivity, whereas asparagine limitation had no significant effect on either growth or cell specific productivity. Neither glutamine nor asparagine limitation significantly affected antibody glycosylation. Replenishing the CHO‐DXB11 culture with cysteine after 1 day of cysteine limitation allowed the cells to partially recover their growth and productivity. This recovery was not observed after 2 days of cysteine limitation. Based on these findings, a fed‐batch protocol was developed using single or mixed amino acid supplementation. Although cell density and antibody concentration were lower compared to a commercial feed, the feeds based on cysteine supplementation yielded comparable cell specific productivity. Overall, this study showed that different amino acid limitations have varied effects on the performance of CHO cell cultures and that maintaining cysteine availability is a critical process parameter for the three cell lines investigated.

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“…Since arginine, aspartic acid, asparagine, glutamine are (Table 2). thermolabile they may be regarded as an indicator of thermal induced alteration of ingredients during the compaction process [7][8][9]14]. No significant difference within and between (p N 0.05) recovered amino acids or vitamins concentration of solutions of compacted CHO210 or Feed210 was observed when compared with the concentrations recovered from solution of their respective powdered form (Figs.…”

Section: Composition Of Compacted Vs Powdered Cho210 Media and Feed210mentioning

confidence: 73%

“…Dry powder cell culture media (DPM) used in large scale processes needs to be easy to handle, and should dissolve quickly. In these large scale setups, CCM solutions are often heated or subject to drastic pH changes to increase the powder dissolution rate which can detrimental to, heat labile and pH sensitive components such as riboflavin [7][8][9]. To maintain homogeneity of CCM and to maximize solvent accessible surface area during dissolution, CCM is milled to a small particle size range.…”

Section: Introductionmentioning

confidence: 99%

Compaction of chemically defined cell culture media increases its dissolution rate through an increase of solvent accessible surface area

et al. 2016

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Cultivation of mammalian cells for the generation of therapeutics requires the use of cell culture media which is a defined mixture of nutrients including carbohydrates, amino acids, lipids, vitamins, and inorganic salts. Its composition influences the cell density, yield, and quality of the molecule expressed. These chemically defined cell culture media are manufactured as powders since they offer advantages of storage, handling and stability. To prevent de-mixing of the powders and maintain homogeneity, these powders are milled down to small particle size ranges. However, this small particle size allows for inter-particle forces to impact powder flow properties, and decreases the surface area available to dissolution. Hence a method is required that improves these powder parameters, without altering their composition. A deeper understanding of the dissolution mechanism of multicomponent mixtures like cell culture media is also needed. Here, cell culture media granules, generated from homogeneous powders by roller compaction are evaluated. The granulation process did not alter the composition of the media and properties of solutions generated thereof. The analysis and comparison of the dissolution profile of powder and compacted media enabled the calculation of the actual surface area available to dissolution. The improvement in granule dissolution can be attributed to dispersion and disintegration of the granules on wetting increasing available surface area. Decrease in inter-particle forces in granules also improves flow-ability properties. Surface character and pores on the granules further augments the improvement in dissolution.

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Amino acids in the cultivation of mammalian cells

Cited by 95 publications

References 86 publications

Impact of CHO Metabolism on Cell Growth and Protein Production: An Overview of Toxic and Inhibiting Metabolites and Nutrients

Impact of CHO Metabolism on Cell Growth and Protein Production: An Overview of Toxic and Inhibiting Metabolites and Nutrients

Effects of cysteine, asparagine, or glutamine limitations in Chinese hamster ovary cell batch and fed‐batch cultures

Compaction of chemically defined cell culture media increases its dissolution rate through an increase of solvent accessible surface area

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