Enhancement of transient gene expression by fed-batch culture of HEK 293 EBNA1 cells in suspension |
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Authors: | Xiangming Sun Peen Ern Goh Kathy T K Wong Tetsushi Mori Miranda G S Yap |
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Institution: | (1) Bioprocessing Technology Institute, Centros, 138668 Singapore, Singapore |
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Abstract: | Enhanced green fluorescence protein (GFP) and erythropoietin (EPO) were used as reporters to assess and improve transient
gene expression in HEK 293 EBNA1 cells. The production of EPO only lasted 3 days and reached 18.1 mg/l in suspension cultures
in 1 l batch bioreactors. However, GFP expression examined in well-plate experiments persisted for 12 days in transfected
cells but decreased rapidly within the next 15 days. These results suggest that the retaining of a plasmid in cells may not
be a limiting factor for protein expression in large-scale transient transfection. To improve cell maintenance and protein
expression, a fed-batch culture was performed using an enriched medium, a mixture of equal volumes of 293 SFM II medium and
a 5 × amino acid solution prepared based on DMEM/F12 medium formula. EPO reached 33.6 mg/l, representing 86% increase over
that of the batch culture. Moreover, the total amount of EPO produced was increased by 165% in view of the volume increase
in the fed-batch culture. The serum-free medium used in this work enables cells growing well and transfection without medium
change. Thus, the process reported here is simple and easy to scale up. |
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Keywords: | HEK 293 EBNA1 cells Polyethylenimine Transient gene expression |
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