transient gene expression in hek293 and vero cells immobilised on microcarriers：在hek293细胞和vero细胞微载体固定化对瞬时基因表达.pdfVIP

Journal of Biotechnology 153 (2011) 15–21 Contents lists available at ScienceDirect Journal of Biotechnology j o u rn al h omepage: w w w. elsev /locate/jbiotec Transient gene expression in HEK293 and vero cells immobilised on microcarriers Lukas Fliedl a , Christian Kaisermayer b,∗ a Institute of Applied Microbiology, Muthgasse 18, 1190 Vienna, Austria b GE Healthcare, Institute of Applied Microbiology, Muthgasse 18, 1190 Vienna, Austria a r t i c l e i n f o a b s t r a c t Article history: The upscale of transient gene expression (TGE) gained popularity over the last decade as it drastically Received 21 September 2010 shortens timelines for the production of recombinant proteins. Bottlenecks of the method turned out Received in revised form 3 February 2011 to be media composition and media exchange, which is usually required as conditioned medium dras- Accepted 21 February 2011 tically reduces the transfection efficiency. Media exchanges are typically done by centrifugation, which Available online 26 February 2011 limits upscale, is prone to contamination or is a high cost factor when continuous centrifuges are used. In this work HEK/EBNA cells were grown and transfected on microcarriers. Cell immobilisation allows easy Keywords: media exchange after sedimentation. The transfection method was optimised regarding polyethylen- Transient gene expression HEK293