title: Recombinant protein production using the Semliki Forest Virus expression system.
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).
authors.affiliation: Glaxo Institute for Molecular Biology, 14 Chemin des Aulx, CH 1228, Plan les Ouates, Switzerland.
authors.forename: H D
authors.initials: HD
authors.lastname: Blasey
authors.forename: K
authors.initials: K
authors.lastname: Lundström
authors.forename: S
authors.initials: S
authors.lastname: Tate
authors.forename: A R
authors.initials: AR
authors.lastname: Bernard