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.
Kompier
R
R
Department of Biophysics, Weizmann Institute of Science, Rehovot, Israel.
Kislev
N
N
Segal
I
I
Kadouri
A
A
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