MRC National Institute for Medical Research, London
Science for Health
A better method for recombinant protein production
17 April 2013
NIMR scientists have developed a novel methodology to produce milligram quantities of highly pure, catalytically active recombinant protein complex. The research is published in Biochemical Journal.
Biological research often requires quantities of proteins to be produced. HEK-293 cells are commonly used for expression of recombinant proteins due to their efficient transient transfection and ability to grow in suspension cultures. However, yields achievable in these cells are far lower than possible with E. coli and baculovirus expression systems. A major problem with the expression of recombinant proteins in mammalian cells is DNA plasmid activation of the cellular virus defence protein PKR (protein kinase RNA-activated) that inhibits protein translation.
Steve Ley (pictured) and his lab, in NIMR’s Division of Immune Cell Biology, have shown that by using the Ebola virus protein VP35 to block PKR activation by plasmid transfection, they can increase protein expression of the protein they were working on, recombinant TPL-2/NF-κB1 p105/ABIN-2, more than ten-fold. This allows purification of sufficient TPL-2 protein complex for biophysical and structural studies. They show that VP35 similarly enhances the expression of co-transfected CD40, TLR4 and Bcl-2, suggesting that method can be used generally for expression of recombinant proteins in HEK-293 cells.
By blocking activation of PKR, we show that Ebola virus VP35 substantially increased the yield of co-expressed recombinant proteins. This simple approach has potentially broad implications for structural studies and drug screening, which require large amounts of purified recombinant protein.
Steve Ley
The article has been selected for F1000Prime. It was
recommended as being of special significance in its field by F1000Prime Faculty
Member Dario Alessi. 
Original article
Thorsten Gantke, Sabrina Boussouf , Julia Janzen, Nick Morrice, Steven Howell, Elke Mühlberger & Steven C. Ley (2013)
Ebola virus VP35 induces high-level production of recombinant TPL-2/ABIN-2/NF-κB1 p105 complex in co-transfected HEK-293 cells
Biochemical Journal. Epub ahead of print. Publisher abstract
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