The response of virally infected insect cells to dissolved oxygen concentration: Recombinant protein production and oxidative damage

• Published in: Biotechnology and bioengineering 2003-01, Vol.81 (1), p.106-114
• Main Authors: Saarinen, Mark A., Murhammer, David W.
• Format: Article
• Language: English
• Subjects: Animals; baculovirus; BTI-Tn-5B1-4; dissolved oxygen; insect cell; lipid oxidation; Oxidative Stress; Oxygen - metabolism; protein oxidation; recombinant protein; Recombinant Proteins - biosynthesis; Sf-9; Spodoptera - cytology; Spodoptera - metabolism; Spodoptera - virology; Thiobarbituric Acid Reactive Substances - metabolism
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.10460

The effects of dissolved oxygen (DO) concentration on virally infected insect cells were investigated in 3‐L bioreactor culture. Specifically, cultures of Spodoptera frugiperda Sf‐9 (Sf‐9) and Trichoplusia ni BTI‐Tn‐5B1‐4 (Tn‐5B1‐4) were infected with Autographa californica multiple nucleopolyhedrovirus expressing secreted alkaline phosphatase (SEAP). Following infection at a DO concentration of 50% air saturation, the DO concentration was adjusted to a final value of either 190%, 50%, or 10% air saturation. Recombinant SEAP production, cell viability, protein carbonyl content, and thiobarbituric acid reactive substances (TBARS) content were monitored. The increases in protein carbonyl and TBARS contents are taken to be indicators of protein oxidation and lipid oxidation, respectively. DO concentration was found to have no noticeable effect on SEAP production or cell viability decline in the Sf‐9 cell line. In the Tn‐5B1‐4 cell line, cells displayed an increased peak SEAP production rate for 190% air saturation and displayed an increased rate of viability decline at increased DO concentration. Protein carbonyl content showed no significant increase in the Sf‐9 cell line by 72 h postinfection (pi) at any DO concentration but showed a twofold increase at 10% and 50% DO concentration and a threefold increase at 190% DO concentration by 72 h pi in Tn‐5B1‐4 cells. TBARS content was found to increase by ≈ 50% in Sf‐9 cells and by ≈ twofold in Tn‐5B1‐4 cells by 72 h pi with no clear relationship to DO concentration. It is hypothesized that oxygen uptake changes due to the viral infection process may bear a relation to the observed increases in protein and lipid oxidation and that lipid oxidation may play an important role in the death of virally infected insect cells. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 106–114, 2003.