Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 108, Issue 3, Pages 600-610Publisher
WILEY
DOI: 10.1002/bit.22968
Keywords
lentiviral vectors; CHO cells; recombinant protein production; stable expression; high producers
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Funding
- Ecole Polytechnique Federale de Lausanne
- Swiss Commission for Technology and Innovation (CTI) [10203.1PFLS-LS]
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Lentivirus-derived vectors (LVs) were studied for the generation of stable recombinant Chinese hamster ovary (CHO) cell lines. Stable pools and clones expressing the enhanced green fluorescent protein (eGFP) were selected via fluorescence-activated cell sorting (FACS). For comparison, cell pools and cell lines were also generated by transfection, using the LV transfer plasmid alone. The level and stability of eGFP expression was greater in LV-transduced cell lines and pools than in those established by transfection. CHO cells were also infected at two different multiplicities of infection with an LV co-expressing eGFP and a tumor necrosis factor receptor:Fc fusion protein (TNFR:Fc). At 2-day post-infection, clonal cell lines with high eGFP-specific fluorescence were recovered by FACS. These clones coexpressed TNFR: Fc with yields of 50-250 mg/L in 4-day cultures. The recovered cell lines maintained stable expression over 3 months in serum-free suspension culture without selection. In conclusion, LV-mediated gene transfer provided an efficient alternative to plasmid transfection for the generation of stable and high-producing recombinant cell lines. Biotechnol. Bioeng. 2011; 108: 600-610. (C) 2010 Wiley Periodicals, Inc.
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