Optimization of Growth Methods and Recombinant Protein Production in BTI-Tn-5B1-4 Insect Cells Using the Baculovirus Expression System

摘要: A novel insect cell line from Trichoplusia ni, BTI-Tn 5B1-4 (Tn 5), was compared to Spodoptera frugiperda, Sf 9, cells for production of two recombinant secreted proteins: truncated Epstein-Barr viral attachment protein (EBV gp105) and truncated, soluble tissue factor (sTF). Under optimum conditions both lines, Tn 5 produced 28-fold more sTF than 9 cells, respectively, on a per basis. The total gp105 similar the lines. However, Tn5 much efficiently, resulting in 5-fold higher levels extracellular medium. Despite these increases, are attachment-dependent, is sensitive density (cells/cm2), unlike Sf9 which can be easily grown scaled up suspension cultures without significantly affecting its production. Thus, above 0.1L scales optimized with respect using standard techniques growth attachment-dependent cells. Roller bottles precoated DEAE-based microcarriers employing collagen-coated were found effective ways culturing Predetermined optimal densities used produce EBV microcarrier-coated roller or at concentrations close those observed culture flasks.