A two-stage bioreactor system for the production of recombinant proteins using a genetically engineered baculovirus/insect cell system.

Abstract

A two-stage bioreactor scheme was developed for the large-scale production of recombinant proteins using a genetically engineered baculovirus/insect cell system. The first bioreactor was employed for cell growth and the second for cell infection. Silkworm Bm5 cells were infected with a recombinant baculovirus, BmNPV/P5.cat, containing a bacterial chloramphenicol acetyltransferase (CAT) gene under the control of the polyhedrin gene promoter of Bombyx mori nuclear polyhedrosis virus (BmNPV). This recombinant baculovirus has been used as an expression vector for the production of recombinant CAT enzyme. A specific productivity of 82 to 90 microg CAT/(10(6) cells) was obtained using the BmNPV/Bm5 expression system, a yield similar to that achieved using the AcNPV/Sf expression system. Repeated infection of high-density cell cultures did not reduce the specific productivity of the CAT enzyme. Most importantly, the problems associated with the infection of high-density cell cultures were resolved by means of controlled infection conditions and appropriate replenishment of spent culture medium following infection. The glucose uptake rate by the cells following infection was 50% higher than that by the cells before infection. Not only did the infection of high-density cell cultures result in consistent yields of 250 mg/L of CAT enzyme, but also the two-stage bioreactor system was proven to be reliable for a long-term operation beyond 600 h.

• Authors: J Zhang, N Kalogerakis, L A Behie, K Iatrou
• Journal: Biotechnology and bioengineering (Biotechnol Bioeng) Vol. 42 Issue 3 Pg. 357-66 (Jul 1993) ISSN: 0006-3592 [Print] United States
• PMID: 18613020 (Publication Type: Journal Article)
• Copyright: (c) 1993 John Wiley & Sons, Inc.