FAS2FURIOUS: Moderate-Throughput Secreted Expression of Difficult Recombinant Proteins in Drosophila S2 Cells

Frontiers in Bioengineering and Biotechnology

Michael Fairhead¹  Nicola A. Burgess-Brown¹  Wyatt W. Yue¹  Vittorio L. Katis¹  Opher Gileadi¹  Willem A. de Jongh²  Stine B. Clemmensen²  Bent U. Frandsen²  Anja Schwenzer³  Brian D. Marsden³  Jesse A. Coker³  Kim S. Midwood³ 

[1] Centre for Medicines Discovery, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom;ExpreS2ion Biotechnologies, SCION-DTU Science Park, Hørsholm, Denmark;Kennedy Institute of Rheumatology, University of Oxford, Oxford, United Kingdom;

关键词: insect cells;    recombinant proteins;    S2 cells;    glycoproteins;    secreted proteins;    structural biology;   

DOI  :  10.3389/fbioe.2022.871933

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown