期刊
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
卷 10, 期 -, 页码 -出版社
FRONTIERS MEDIA SA
DOI: 10.3389/fbioe.2022.871933
关键词
insect cells; recombinant proteins; S2 cells; glycoproteins; secreted proteins; structural biology
Recombinant protein expression in eukaryotic insect cells, particularly using Drosophila melanogaster S2 cells, has great potential for challenging targets. The development of FAS2FURIOUS provides a simple and rapid platform for cloning and testing multiple constructs in a short period, making it an attractive orthogonal approach for protein expression in eukaryotic cells.
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 (TM). 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.
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