Recombinant protein production using the Semliki Forest Virus expression system.

Cytotechnology
H D BlaseyA R Bernard

Abstract

We report here the successful scale up of transient recombinant protein expression to litre scale using Semliki Forest Virus System. The expression of bacterial β-galactosidase was initially compared in BHK and CHO cells and the conditions for optimal infection of BHK cells were identified. 10% FCS in a medium at pH 6.9 and infection in small volumes were found to be optimal. A high MOI results in an increased recombinant protein yield. Stirring does not affect the infection process. Finally we applied these optimal conditions to the production of a microsomal enzyme, human cyclooxygenase-2 in suspension spinners. Five independant productions at the 1 litre scale yielded reproducible substantial amounts of recombinant protein (16 mg microsomal protein 10(9) cells(-1)) with an average specific activity of 3942 ± 765 pg PGE(2) μg(-1) microsomal protein 5 min(-1).

Citations

Nov 23, 2006·Journal of Structural and Functional Genomics·Kenneth LundstromFranc Pattus
Apr 25, 2003·Journal of Virological Methods·Seema S GangolliJacek Kowalski
Nov 14, 1997·Current Opinion in Biotechnology·K Lundstrom
Apr 21, 1999·Current Opinion in Biotechnology·F Wurm, A Bernard
Apr 12, 2013·Future Medicinal Chemistry·Kenneth Lundstrom
Jul 6, 2016·Molecular Therapy : the Journal of the American Society of Gene Therapy·Juliane W SchottAxel Schambach
Jan 20, 2007·Explore : the Journal of Science and Healing·Heinz Birnesser, Pelle Stolt
Nov 6, 2001·Journal of Receptor and Signal Transduction Research·K LundstromF Jenck
Dec 31, 2002·Journal of Receptor and Signal Transduction Research·Kenneth Lundstrom
Mar 11, 1999·Journal of Receptor and Signal Transduction Research·K Lundstrom

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