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1 October 2001 DESIGN OF AN EFFICIENT MEDIUM FOR INSECT CELL GROWTH AND RECOMBINANT PROTEIN PRODUCTION
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Abstract

We report the development of a new serum-free medium based on the use of factorial experiments. At first, a variety of hydrolysates were screened using a fractional factorial approach with High-Five cells. From this experiment yeastolate ultrafiltrate was found to have, by far, the most important effect on cell growth. Furthermore, Primatone RL® was found to remarkably prolong the stationary phase of Sf-9 and High-Five cell cultures. The optimal concentrations for yeastolate and Primatone were determined to be 0.6 and 0.5%, respectively, on the basis of a complete factorial experiment. This new medium, called YPR, supported good growth of both Sf-9 and High-Five cells in batch cultures, with maximal densities of 5.4 and 6.1 × 106 cells/ml, respectively. In addition, both cell lines achieved good growth in bioreactor batch culture and had a prolonged stationary phase of 3–4 d in YPR medium compared to Insect-XPRESS medium. The ability of the new medium to support recombinant protein expression was also tested by infecting Sf-9 or High-Five cells at high density (2 × 106 cells/ml) with a baculovirus expressing secreted placental alkaline phosphatase (SEAP). The maximum total SEAP concentration after 7 d was about 43 IU/ml (58 mg/L) and 28 IU/ml (39 mg/L) for High-Five and Sf-9 cells, respectively.

L. IKONOMOU, G. BASTIN, Y.-J. SCHNEIDER, and S. N. AGATHOS "DESIGN OF AN EFFICIENT MEDIUM FOR INSECT CELL GROWTH AND RECOMBINANT PROTEIN PRODUCTION," In Vitro Cellular & Developmental Biology - Animal 37(9), 549-559, (1 October 2001). https://doi.org/10.1290/1071-2690(2001)037<0549:DOAEMF>2.0.CO;2
Received: 8 June 2001; Published: 1 October 2001
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