Synthechol™ Ynthetic Cholesterol for Cholesterol Dependent Cell Culture — Development of Non-Animal Derived Chemically Defined NS0 Medium

Talley, D.; Cutak, Benjamin J.; Rathbone, Elner B.; Kolla, T. Al; Allison, Daniel W.; Blasberg, Justin D.; Kao, Kung-Ying Tang; Caple, Matthew V.

doi:10.1007/1-4020-3103-3_114

Cited by 6 publications

(4 citation statements)

References 1 publication

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“…The replacements were either the recombinant/plant counterparts or other molecules with similar function. Specifically, bovine serum albumin could be replaced by recombinant human albumin, and also by an organic carrier for lipids, cyclodextrin (Jayme, 1999;Zhang and Robinson, 2005); bovine lipoprotein by synthetic cholesterol and plant-derived fatty acids (Gorfien et al, 2000;Talley et al, 2003), bovine insulin by recombinant human insulin or zinc ions (Wong et al, 2004;Zhang and Robinson, 2005); and bovine transferrin by small molecule tropolone (2-hydroxy-2,4, 6-cycloheptatrien-1) for iron delivery (Metcalfe et al, 1994) or other iron-chelating agents (Schenerman et al, 2003). Iron is essential for cell respiration and metabolism, and without iron cells stop growing and eventually die.…”

Section: Introductionmentioning

confidence: 99%

A novel function for selenium in biological system: Selenite as a highly effective iron carrier for Chinese hamster ovary cell growth and monoclonal antibody production

Zhang

Robinson

Salmon

2006

Biotechnol. Bioeng.

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As the market for biopharmaceuticals especially monoclonal antibodies (MAbs) rapidly grows, their manufacturing methods are coming under increasing regulatory scrutiny, particularly due to concerns about the potential introduction of adventitious agents from animal-sourced components in the media used for their production in mammalian cell culture. Chinese hamster ovary (CHO) cells are by far the most commonly used production vehicles for these recombinant glycoproteins. In developing animal-component free media for CHO and other mammalian cell lines, the iron-transporter function of serum or human/bovine transferrin is usually replaced by certain organic or inorganic chelators capable of delivering iron for cell respiration and metabolism, but few of them are sufficiently effective. Selenium is a well-known essential trace element (TE) for cell growth and development, and its positive role in biological system includes detoxification of free radicals by activating glutathione peroxidase. In cell culture, selenium in the form of selenite can help cells to detoxify the medium thus protect them from oxidative damage. In this presentation, we describe the discovery and application of a novel function of selenite, that is, as a highly effective carrier to deliver iron for cell growth and function. In our in-house-developed animal protein-free (APF) medium for CHO cells, using an iron-selenite compound to replace the well-established tropolone delivery system for iron led to comparable or better cell growth and antibody production. A high cell density of >10 x 10(6) viable cells/mL and excellent antibody titer of approximately 3 g/L were achieved in 14-day fed-batch cultures in shake flasks, followed by successful scale-up to stirred bioreactors. The preparation of the commercially unavailable iron-selenite compound from respective ions, and its effectiveness in cell-culture performance, were dependent on reaction time, substrates, and other conditions.

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Section: Introductionmentioning

confidence: 99%

A novel function for selenium in biological system: Selenite as a highly effective iron carrier for Chinese hamster ovary cell growth and monoclonal antibody production

Zhang

Robinson

Salmon

2006

Biotechnol. Bioeng.

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“…Subsequently, the crosslinked P84 membranes are used for adsorption of Lys in DMSO and cholesterol (Chol) in NMP to demonstrate the adsorption properties of the solvent resistant P84 membranes. Both Lys and Chol play important roles in the mammalian bodies and are widely used in biotechnology [24,[32][33][34]. In the aqueous systems, the adsorption is based on electrostatic interactions between the biomolecules and charged particles whereas in the organic solvents hydrophobic/hydrophilic interactions play a dominant role.…”

Section: Introductionmentioning

confidence: 99%

Solvent-resistant P84-based mixed matrix membrane adsorbers

Kopec

Dutczak

Bolhuis-Versteeg

et al. 2011

Separation and Purification Technology

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“…Although progress has been made to culture recombinant NS0 cell lines at a large scale in animal component free, chemically defined media, significant opportunities exist to optimize the base medium, medium supplements and culturing protocols to improve both cell growth and mAb productivity. 3, 4–9…”

Section: Introductionmentioning

confidence: 99%

Enhanced productivity of NS0 cells in fed‐batch culture with cholesterol nanoparticle supplementation

Wyslouzil

et al. 2011

Biotechnology Progress

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NS0 cells are an important industrial cell line for the production of therapeutic monoclonal antibodies. Culturing these cells is challenging because they are cholesterol auxotrophs, and providing cholesterol to the cells is hampered by the low solubility of lipids in aqueous medium. Limited loading capacity, precipitation, instability, and toxicity are associated with traditional delivery methods that involve solvents or carrier molecules. In this work, nanoparticle cholesterol mixtures (NCM) were produced by electrohydrodynamic spraying and added directly to a cholesterol auxotroph NS0 cell line. Compared to a cholesterol-cyclodextrin solution and a commercial proprietary cholesterol solution, SyntheChol NS0 supplement, NCM is significantly less cytotoxic. In the fed batch cell culture process, product titer was increased by 32% when the NCM supplement replaced SyntheChol NS0 supplement. An even greater product titer improvement, 64%, was achieved when both NCM and SyntheChol NS0 supplements were used in the fed-batch process.

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Synthechol™ Ynthetic Cholesterol for Cholesterol Dependent Cell Culture — Development of Non-Animal Derived Chemically Defined NS0 Medium

Cited by 6 publications

References 1 publication

A novel function for selenium in biological system: Selenite as a highly effective iron carrier for Chinese hamster ovary cell growth and monoclonal antibody production

A novel function for selenium in biological system: Selenite as a highly effective iron carrier for Chinese hamster ovary cell growth and monoclonal antibody production

Solvent-resistant P84-based mixed matrix membrane adsorbers

Enhanced productivity of NS0 cells in fed‐batch culture with cholesterol nanoparticle supplementation

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