Use of a stationary bed reactor and serum-free medium for the production of recombinant proteins in insect cells.
Insect cells (Spodoptera frugiperda) have been cultured in a stationary bed reactor, packed with a fibrous polyester carrier. When the bioreactor was perfused with serum-supplemented medium, a cell density of 6 x 10(6) cells ml-1 packed carrier was reached. Scanning electron microscopy investigations have shown that the insect cells grew along the three-dimensionally oriented fibers of the Fibra-cel carrier. After infection of the logarithmically growing cells with a recombinant baculovirus (Autographa californica) containing the gene coding for beta-galactosidase, the medium in the bioreactor was changed to serum-free medium. At day 13 postinfection (p.i.), a beta-galactosidase level of 320 microgram ml-1 and, at day 17 p.i., a virus titer of 2.1 x 10(8) TCID50 units ml-1 (day 17 p.i.) were reached. In another bioreactor, operated in a similar way but with serum-containing medium, a beta-galactosidase concentration of 360 microgram ml-1 and a virus titer of 2.3 x 10(8) TCID50 units ml-1 were obtained. These results indicate the potential use of this production system for the production of recombinant protein and baculovirus in insect cells.
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
R
R
Kompier
N
N
Kislev
I
I
Segal
A
A
Kadouri
D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000191:economics / Q000295:instrumentation / Q000379:methods*; D002455:Cell Division; D002460:Cell Line; D003365:Costs and Cost Analysis; D016895:Culture Media, Serum-Free; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D009036:Moths; D011994:Recombinant Proteins / Q000096:biosynthesis*; D001616:beta-Galactosidase / Q000235:genetics
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D001616:beta-Galactosidase