Effect of lipids on insect cell growth and expression of recombinant proteins in serum-free medium
โ Scribed by Rebecca S. Gilbert; Yuichi Nagano; Tadafumi Yokota; Sun Fon Hwan; Tom Fletcher; Ken Lydersen
- Publisher
- Springer
- Year
- 1996
- Tongue
- English
- Weight
- 342 KB
- Volume
- 22
- Category
- Article
- ISSN
- 0920-9069
No coin nor oath required. For personal study only.
โฆ Synopsis
The lipid emulsion components of a serum-free insect cell medium were varied and evaluated for effects on cell growth and recombinant protein expression. The growth of High-Five(TM) cells was significantly affected by polyol Pluronic F-68 and Tween-80, but not by lipids. Pluronic was essential for cell growth, while Tween-80 was required to achieve maximum cell densities. A dose response effect was observed for Tween-80 with optimal cell growth at a concentration of 25 mg/l. Cholesterol had a minor effect on cell growth, but was essential for the expression of recombinant proteins. The expression of ฮฒ-galactosidase (ฮฒ-gal) was directly affected by cholesterol with optimal expression at a concentration of 5.4 mg/l. Vitamin E, important as an antioxidant to stabilize lipids, did not directly affect recombinant protein expression. Although lipids were not required for cell growth, the presence of lipids were required during the cell growth phase in order to achieve efficient infection with baculovirus. These studies help to define the important components, and range of concentrations, for lipid emulsions which can effectively replace serum in insect cell culture.
๐ SIMILAR VOLUMES
The effect of temperature and Oz saturation on the production of recombinant proteins fl-galactosidase and human glucocerebrosidase by Spodoptera frugiperda cells (Sf9) infected with recombinant Autographa californica nuclear polyhedrosis virus was investi- gated. The rates of cell growth, glucose c
BiP, GRP94 and PDI, three endoplasmic reticulum (ER) based proteins are involved in the maturation of secretory proteins and might represent a bottleneck in the secretory pathway of monoclonal antibodies (MAB). With the three hybridoma cell lines tested, MAB production kinetics were significantly in
Medium that had supported the growth of insect cells was depleted in components of both the basal medium and foetal calf serum. In addition, a non-dialysable growth-inhibiting factor(s) appeared to exist. The pH had risen to a sub-optimal level and simply readjusting this to pH 6.6 partially restore