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

  1. Get@NRC: Serum-free production of recombinant proteins and adenoviral vectors by 293SF-3F6 cells (Opens in a new window)
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Journal titleBiotechnology and Bioengineering
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
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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