Serum-free production of recombinant proteins and adenoviral vectors by 293SF-3F6 cells

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TypeArticle
Journal titleBiotechnology and Bioengineering
Volume59
Issue5
Pages567575; # of pages: 9
Subjectadenovirus; adenoviral vectors; 293 cells; recombinant proteins; serum-free medium
AbstractThis article describes the step-wise approach undertaken to select a serum-free medium (SFM) for the efficient production of a recombinant adenoviral vectors expressing β-galactosidase (Ad5 CMV-LacZ), in the complementing human embryonic kidney 293S cells. In the first step, a 293S-derived transfectoma, secreting a soluble epidermal growth factor receptor sEGFr (D2-22), was used to estimate the potential of selected serum-free formulations to support the production of a recombinant protein as compared to serum-containing medium. Assays showed that only one among six commercial serum-free formulations could support both sEGFr production and cell growth in static or suspension culture. In commercially available calcium-containing serum-free formulations, the cell aggregates reached up to 3 mm in diameter. In the second step, 293S cells were gradually adapted to a low-calcium version of the selected medium (LC-SFM). Cells were cloned, then screened according to their ability to grow at a rate and an extent comparable to parental cells in serum-containing medium (standard) as single cells or small aggregates. The 293SF-3F6 clone, first adapted to and then cloned in the selected serum-free medium, was selected for further experiments. Bioreactor run performed with the 293SF-3F6 clone showed similar growth curve as in the shake-flask controls. In the final step, the recombinant viral vector productivity of the 293S cells and the 293SF-3F6 clone was tested. The 293SF-3F6 cells infected by Ad5 CMV-LacZ in 3 L-scale bioreactor maintained the specific productivities of both β-galactosidase and adenoviral vector equivalent to the shake-flask controls in suspension culture. Results from this study clearly demonstrate that the 293SF-3F6 cell line thus selected may be used either for establishing stable transfected cell line or for the production of adenoviral vectors required for gene therapy studies.
Publication date
Linkhttp://onlinelibrary.wiley.com/doi/10.1002/(SICI)1097-0290(19980905)59:5%3C567::AID-BIT6%3E3.0.CO;2-8/abstract
LanguageEnglish
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
NoteUI - 99201169
Peer reviewedNo
NRC number41493
NPARC number3539982
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Record identifierf7bcd54b-8018-4df1-85a7-6f7e85c516b5
Record created2009-03-01
Record modified2016-05-09
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