Optimization of protein-production by the baculovirus expression vector system in shake flasks.
Shake flasks were successfully employed for the cultivation of Spodoptera frugiperda (Sf-9) insect cells and for the production of beta-galactosidase, a recombinant model protein, utilizing the baculovirus expression vector system. The culture doubling time and maximal cell density were 20 h and 5 x 10(6) cells/ml respectively. The optimal liquid volumes for flasks rotating at 100 rpm were 25-40% of the flask total volume. Enzyme production (about 600 mg/l) was best at a multiplicity of infection of between 1 and 20 and at a cell density at time of infection of 0.7 x 10(6) cells/ml. At a rotation speed of 100 rpm, Pluronic F-68 had no effect on growth and enzyme production.
Department of Food Engineering and Biotechnology, The Technion, Technion City, Haifa, Israel.
R
R
Neutra
B Z
BZ
Levi
Y
Y
Shoham
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002478:Cells, Cultured; D005822:Genetic Vectors; D007313:Insecta / Q000166:cytology; D011994:Recombinant Proteins / Q000096:biosynthesis; D001616:beta-Galactosidase / Q000096:biosynthesis*
D011994:Recombinant Proteins; D001616:beta-Galactosidase