1. FAS2FURIOUS: Moderate-Throughput Secreted Expression of Difficult Recombinant Proteins in Drosophila S2 Cells
- Author
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Jesse A. Coker, Vittorio L. Katis, Michael Fairhead, Anja Schwenzer, Stine B. Clemmensen, Bent U. Frandsen, Willem A. de Jongh, Opher Gileadi, Nicola A. Burgess-Brown, Brian D. Marsden, Kim S. Midwood, and Wyatt W. Yue
- Subjects
Histology ,Biomedical Engineering ,Bioengineering ,Biotechnology - Abstract
Recombinant protein expression in eukaryotic insect cells is a powerful approach for producing challenging targets. However, due to incompatibility with standard baculoviral platforms and existing low-throughput methodology, the use of the Drosophila melanogaster “S2” cell line lags behind more common insect cell lines such as Sf9 or High-Five™. Due to the advantages of S2 cells, particularly for secreted and secretable proteins, the lack of a simple and parallelizable S2-based platform represents a bottleneck, particularly for biochemical and biophysical laboratories. Therefore, we developed FAS2FURIOUS, a simple and rapid S2 expression pipeline built upon an existing low-throughput commercial platform. FAS2FURIOUS is comparable in effort to simple E. coli systems and allows users to clone and test up to 46 constructs in just 2 weeks. Given the ability of S2 cells to express challenging targets, including receptor ectodomains, secreted glycoproteins, and viral antigens, FAS2FURIOUS represents an attractive orthogonal approach for protein expression in eukaryotic cells.
- Published
- 2022
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