CULTURE IN THE ROTATING-WALL VESSEL AFFECTS RECOMBINANT PROTEIN PRODUCTION CAPABILITY OF TWO INSECT CELL LINES IN DIFFERENT MANNERS

• Published in: In vitro cellular & developmental biology. Animal 2000-06, Vol.36 (6), p.362-366
• Main Authors: SAARINEN, MARK A, MURHAMMER, DAVID W
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - biosynthesis; Animals; baculovirus expression vector system; Bioreactors; BIOTECHNOLOGY; Cell aggregates; Cell Aggregation; Cell culture techniques; Cell Culture Techniques - instrumentation; Cell Culture Techniques - methods; Cell Line; Cell lines; Cell Survival; Cultured cells; Flasks; Glucose - metabolism; high aspect rotating-wall vessel; Insect proteins; Insect vectors; Lactates; Lactates - metabolism; Moths; Nucleopolyhedrovirus - physiology; Recombinant proteins; Recombinant Proteins - biosynthesis; Rotation; secreted alkaline phosphatase; Sf-9; Space life sciences; Spodoptera; Spodoptera frugiperda; Stress, Mechanical; Trichoplusia ni; Trichoplusia ni BTI-Tn-5B1-4; Viability
• ISSN: 1071-2690; 1543-706X; 1543-706X
• DOI: 10.1290/1071-2690(2000)036<0362:CITRWV>2.0.CO;2

The production of recombinant secreted alkaline phosphatase protein in virally infected insect cells was studied in shaker flask and high aspect rotating-wall vessel (HARV) culture. Two commonly used cell lines, Spodoptera frugiperda Sf-9 (Sf-9) and a nonaggregating isolate of the Trichoplusia ni BTI-Tn-5B1-4 (Tn-5B1-4) cell line, Trichoplusia ni Tn-5B1-4-NA (Tn-5B1-4-NA), were used and monitored for 120-h postinfection. Different responses to culture in the HARV were seen in the two cell lines. While the Sf-9 cell line was able to produce slightly greater amounts of recombinant protein in the HARV than in shaker flask controls, the Tn-5B1-4-NA cell line produced significantly lesser amounts in the HARV than in the shaker flasks. Both cell lines exhibited longer life spans and longer periods of protein production in HARV culture than in shaker flask culture, presumably due to lower levels of shear encountered in the HARV. The important difference was in the protein production rate responses of the two cell lines. While the protein production rates of Sf-9 cells were comparable in both HARV and shaker flask cultures, the protein production rates of Tn-5B1-4-NA cells were much lower in HARV culture than in shaker flask cultures. The conclusion is drawn that cell line–specific adaptation to the HARV strongly influences recombinant protein production.