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Fed-batch productio...
Fed-batch production assessment of a tetravalent bispecific antibody : A case study on piggyBac stably transfected HEK293 cells
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- Napoleone, Antonino (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Science for Life Laboratory, SciLifeLab
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- Lauren, Ida (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Science for Life Laboratory, SciLifeLab
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- Linkgreim, Triinu (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Science for Life Laboratory, SciLifeLab
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- Dahllund, Leif (författare)
- KTH,Läkemedelsutveckling,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Sci Life Lab, Drug Discovery & Dev, Stockholm, Sweden.;KTH Royal Inst Technol, Sch Engn Sci Chem Biotechnol & Hlth, Stockholm, Sweden.
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- Persson, Helena (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Proteinteknologi,KTH Royal Inst Technol, Sci Life Lab, Drug Discovery & Dev, Stockholm, Sweden.;KTH Royal Inst Technol, Sch Engn Sci Chem Biotechnol & Hlth, Stockholm, Sweden.
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- Andersson, Oskar (författare)
- KTH,Läkemedelsutveckling,Science for Life Laboratory, SciLifeLab,KTH Royal Inst Technol, Sci Life Lab, Drug Discovery & Dev, Stockholm, Sweden.;KTH Royal Inst Technol, Sch Engn Sci Chem Biotechnol & Hlth, Stockholm, Sweden.
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- Olsson, Anders (författare)
- KTH,Science for Life Laboratory, SciLifeLab,Läkemedelsutveckling,KTH Royal Inst Technol, Sci Life Lab, Drug Discovery & Dev, Stockholm, Sweden.;KTH Royal Inst Technol, Sch Engn Sci Chem Biotechnol & Hlth, Stockholm, Sweden.
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- Hultqvist, Greta, 1980- (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Science for Life Laboratory, SciLifeLab
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- Frank, Peter (författare)
- Wicket AB, Aderkvarnsgatan 30, S-75329 Uppsala, Sweden.
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- Hall, Martin (författare)
- Cytiva AB, Bjorkgatan 30, S-75184 Uppsala, Sweden.
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- Morrison, Annika (författare)
- Cytiva Testa Ctr AB, Bjorkgatan 30, S-75184 Uppsala, Sweden.
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- Andersson, Andreas (författare)
- Cytiva Testa Ctr AB, Bjorkgatan 30, S-75184 Uppsala, Sweden.
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- Lord, Martin (författare)
- Uppsala universitet,Institutionen för farmaceutisk biovetenskap,Science for Life Laboratory, SciLifeLab
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- Mangsbo, Sara, 1981- (författare)
- Uppsala universitet,Science for Life Laboratory, SciLifeLab,Institutionen för farmaceutisk biovetenskap
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(creator_code:org_t)
- Elsevier BV, 2021
- 2021
- Engelska.
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Ingår i: New Biotechnology. - : Elsevier BV. - 1871-6784 .- 1876-4347. ; 65, s. 9-19
- Relaterad länk:
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https://doi.org/10.1...
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https://doi.org/10.1...
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https://uu.diva-port... (primary) (Raw object)
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- The transition from preclinical biological drug development into clinical trials requires an efficient upscaling process. In this context, bispecific antibody drugs are particularly challenging due to their propensity to form aggregates and generally produce low titers. Here, the upscaling process for a tetravalent bispecific antibody expressed by a piggyBac transposon-mediated stable HEK293 cell pool has been evaluated. The project was performed as a case study at Testa Center, a non-GMP facility for scale-up testing of biologics in Sweden, and encompassed media adaptation strategies, fed-batch optimization and a novel antibody purification technology. The cell pool was adapted to different culture media for evaluation in terms of cell viability and titers compared to its original Expi293 Expression Medium. These parameters were assessed in both sequential stepwise adaption and direct media exchanges. By this, a more affordable medium was identified that did not require stepwise adaptation and with similar titers and viability as in the Expi293 Expression Medium. Fed-batch optimizations resulted in culture densities reaching up to 20 x 106 viable cells/mL with over 90 % viability 12 days postinoculum, and antibody titers three times higher than corresponding batch cultures. By implementing a novel high-speed protein A fiber technology (Fibro PrismA) with a capture residence time of only 7.5 s, 8 L of supernatant could be purified in 4.5 h without compromising the purity, structural integrity and function of the bispecific antibody. Results from this study related to medium adaptation and design of fed-batch protocols will be highly beneficial during the forthcoming scale-up of this therapeutic antibody.
Ämnesord
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
Nyckelord
- PiggyBac
- HEK293 cell pool
- Bispecific antibody
- Medium adaptation
- Fed-batch
- Fibro PrismA affinity purification
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Napoleone, Anton ...
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Lauren, Ida
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Linkgreim, Triin ...
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Dahllund, Leif
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Persson, Helena
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Andersson, Oskar
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Olsson, Anders
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Hultqvist, Greta ...
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Frank, Peter
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Hall, Martin
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Morrison, Annika
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Andersson, Andre ...
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Lord, Martin
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Mangsbo, Sara, 1 ...
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