Enhanced growth of sf-9 cells to a maximum density of 5.2 × 107 cells per mL and production of β-galactosidase at high cell density by fed batch culture
✍ Scribed by Cynthia B. Elias; Arno Zeiser; Charles Bédard; Amine A. Kamen
- Publisher
- John Wiley and Sons
- Year
- 2000
- Tongue
- English
- Weight
- 207 KB
- Volume
- 68
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
Significant improvement in cell growth and protein production has been achieved in Sf-9 insect cell cultures using pulse additions of multicomponent nutrient feed concentrates (Be ´dard et al., 1994;Chan et al., 1998). The present work focuses on investigating an alternative feeding strategy wherein the nutrients are fed in a semi continuous manner. Fed batch culture experiments were carried out to compare the two different feeding strategies, pulse and semi continuous and a process developed to achieve a cell density of 5.2 × 10 7 cells/ mL of Sf-9 cells in a 3.5 L bioreactor. Production of recombinant protein -galactosidase was carried out by infecting the cells with baculovirus at a MOI of 10 at cell densities of 17 × 10 6 cells/mL. Specific productivity could be maintained at cell densities as high as 14 × 10 6 cells/ mL. The results presented indicate that the feeding method can provide significant improvements in the performance with a reduction in the amount of total nutrients added. On-line monitoring of the culture using the capacitance probe showed that the capacitance probe can be used successfully to monitor the biomass and infection process even at higher cell densities.