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Small-scale bioreac...
Small-scale bioreactor supports high density HEK293 cell perfusion culture for the production of recombinant Erythropoietin
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- Schwarz, Hubert (author)
- KTH,Industriell bioteknologi,Cell Technology Group (CETEG)
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- Zhang, Ye (author)
- KTH,Centrum för Bioprocessteknik, CBioPT
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- Zhan, Caijuan (author)
- KTH,Skolan för teknikvetenskap (SCI),Industriell bioteknologi,Cell Technology Group (CETEG)
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- Malm, Magdalena, 1983- (author)
- KTH,Proteinvetenskap
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- Field, Raymond (author)
- Biopharmaceutical Development, MedImmune, Cambridge, UK
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- Turner, Richard (author)
- Biopharmaceutical Development, MedImmune, Cambridge, UK
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- Sellick, Christopher (author)
- Biopharmaceutical Development, MedImmune, Cambridge, UK
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- Varley, Paul (author)
- Biopharmaceutical Development, MedImmune, Cambridge, UK
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- Rockberg, Johan (author)
- KTH,Proteinteknologi
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- Chotteau, Véronique, Docent, 1963- (author)
- KTH,Skolan för teknikvetenskap (SCI)
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(creator_code:org_t)
- English.
- Related links:
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https://urn.kb.se/re...
Abstract
Subject headings
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- Process intensification in mammalian cell culture-based recombinant protein production has been achieved by high cell density perfusion exceeding 108 cells/mL in the recent years. As the majority of therapeutic proteins are produced in Chinese Hamster Ovary (CHO) cells, intensified perfusion processes have been mainly developed for this type of host cell line. However, the use of CHO cells can result in non-human posttranslational modifications of the protein of interest, which may be disadvantageous compared with human cell lines.In this study, we developed a high cell density perfusion process of Human Embryonic Kidney (HEK293) cells producing recombinant human Erythropoietin (rhEPO). Firstly, a small-scale perfusion system from commercial bench-top screening bioreactors was developed for <250 mL working volume. Then, after the first trial runs with CHO cells, the system was modified for HEK293 cells (more sensitive than CHO cells) to achieve a higher oxygen transfer under mild aeration and agitation conditions. Steady states for medium (20 x 106 cells/mL) and high cell densities (80 x 106 cells/mL), normal process temperature (37 °C) and mild hypothermia (33 °C) as well as different cell specific perfusion rates (CSPR) from 10 to 60 pL/cell/day were applied to study the performance of the culture. The volumetric productivity was maximized for the high cell density steady state but decreased when an extremely low CSPR of 10 pL/cell/day was applied. The shift from high to low CSPR strongly reduced the nutrient uptake rates. The results from our study show that human cell lines, such as HEK293 can be used for intensified perfusion processes.
Subject headings
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
Keyword
- Chinese Hamster Ovary (CHO) cells
- Erythropoietin
- High cell density culture
- Human Embryonic Kidney 239 (HEK293) cells
- Perfusion process
- Small-scale bioreactor
Publication and Content Type
- vet (subject category)
- ovr (subject category)
To the university's database
- By the author/editor
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Schwarz, Hubert
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Zhang, Ye
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Zhan, Caijuan
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Malm, Magdalena, ...
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Field, Raymond
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Turner, Richard
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Sellick, Christo ...
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Varley, Paul
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Rockberg, Johan
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Chotteau, Véroni ...
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- About the subject
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- ENGINEERING AND TECHNOLOGY
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ENGINEERING AND ...
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and Industrial Biote ...
- By the university
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Royal Institute of Technology