Use of a stationary bed reactor and serum-free medium for the production of recombinant proteins in insect cells

• Published in: Enzyme and microbial technology Vol. 13; no. 10; pp. 822 - 827
• Main Authors: Kompier, R., Kislev, N., Segal, I., Kadouri, A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.10.1991; Elsevier Science
• Subjects: Animal cells; Animals; Baculoviridae - genetics; Baculoviridae - metabolism; baculovirus; beta -galactosidase; beta-Galactosidase - genetics; Biological and medical sciences; Bioreactor; bioreactors; Biotechnology; Biotechnology - economics; Biotechnology - instrumentation; Biotechnology - methods; Cell Division; Cell Line; cloning vectors; Costs and Cost Analysis; Culture Media, Serum-Free; Establishment of new cell lines, improvement of cultural methods, mass cultures; Eukaryotic cell cultures; Fundamental and applied biological sciences. Psychology; insect cells; Kinetics; Methods. Procedures. Technologies; Microscopy, Electron, Scanning; Moths; proteins; Recombinant Proteins - biosynthesis; recombinants; serum-free medium; Spodoptera frugiperda; Spodoptera frugiperda, Autographa californica; β-galactosidase; Spodoptera frugiperda, Autographa californica; β-galactosidase; Bioreactor; baculovirus; serum-free medium; insect cells; Growing phase; Cell culture; Scanning electron microscopy; Serum free medium; Packed bed; Enzyme; Insecta; Spodoptera frugiperda; β-D-Galactosidase; Baculovirus; Stationary system; Virus; Infection; Baculoviridae; Arthropoda; Animal; Lepidoptera; Production; Noctuidae; Recombinant protein; Invertebrata
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/0141-0229(91)90066-J

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 × 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 β-galactosidase, the medium in the bioreactor was changed to serum-free medium. At day 13 postinfection (p.i.), a β-glactosidase level of 320 μg ml −1 and, at day 17 p.i., a virus titer of 2.1 × 10 8 TCID 50 units ml −1 (day 17 p.i.) were reached. In another bioreactor, operated in a similar way but with serum-containing medium, a β-galactosidase concentration of 360 μg ml −1 and a virus titer of 2.3 × 10 8 TCID 50 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.