OPTIMIZATION OF RECOMBINANT DENGUE ENVELOPE PROTEIN PRODUCTION IN INSECT CELL CULTURE IN 2.5 L FERMENTER

摘要

The baculovirus expression vector system (BEVS) has been used to express many fuctionally authentic recombinant proteins in insect cells. Insect cells are used as hosts for recombinant baculovirus infection which results in recombinant protein production. In this study, recombinant dengue envelope protein production. In this study, recombinant dengue envelope protein was produced from insect cell (SF-9) infected with recombinant dengue baculovirus. Conditions involve the recombinant dengue envelope protein production i.e insect cell medium, insect cell growth phase, cell density, amout of recombinant baculovirus used to infect cells and optimal harvesting time were investigated. The results obtained when study in shake flask (250ml) showed that optimal medium for Sf-9 growth was the mixture of SF-900II and TC100 at ratio 1:1 supplemented with 10% FCS. The optimal growth phase and cell density were at early log phase and 1x10~6 cells/ml, respectively. Optimal multiplicity of infection was 5 (MOI =5)and harvest time was 3 days post infection. The information obtained in this study was then used for large scale production in a 2.5 L stirred tank reactor performed in 1 L working volume. It was found that the optimal agitation rate and dissolved oxygen for insect cell growth was at 90 rpm and 60-80% air saturation, respectively. Maximum recombinant dengue envelope protein production was 13 mg/L which was higher than the production from shake flask.