Transient transfection of insect Sf-9 cells in TubeSpin®bioreactor 50 tubes

Xiao Shen,Florian M Wurm,Patrik O Michel,Qiuling Xie,David L Hacker

doi:10.1186/1753-6561-5-s8-p37

Xiao Shen, Florian M Wurm + Show 3 more

Open Access

https://doi.org/10.1186/1753-6561-5-s8-p37

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Abstract

BackgroundSf-9 cells, derived fromSpodoptera frugiperda,arewidely used for recombinant protein production usingthe baculovirus expression vector system (BEVS). How-ever, this results in a productive viral infection and celllysis. Therefore, a non-lytic, plasmid-based expressionsystem for suspension Sf-9 cells would be a valuablealternative to the BEVS for rapid, scalable, and high-yielding recombinant protein production and for thegeneration of stable Sf-9 cell lines [4]. In this work, wepresent a simple, efficient and cost-effective plasmid-based method for transient expression of recombinantproteins in Sf-9 cells cultivated in serum-free suspensionmode in a high-throughput culture system, TubeSpin

Highlights

Sf-9 cells, derived from Spodoptera frugiperda, are widely used for recombinant protein production using the baculovirus expression vector system (BEVS)
The human tumor necrosis factor receptor-Fc fusion protein (TNFR-Fc) gene was cloned into pIEx10 (Novagen, Merck, Darmstadt, Germany) to generate pIEx-TNFRFc
The day, the cells were transfected with 1.5 μg pTNFR-Fc and 2.25 μg linear 25 kDa polyethylenimine (PEI, Polysciences, Warrington, PA) per 106 cells

Summary

Background

Sf-9 cells, derived from Spodoptera frugiperda, are widely used for recombinant protein production using the baculovirus expression vector system (BEVS). This results in a productive viral infection and cell lysis. A non-lytic, plasmid-based expression system for suspension Sf-9 cells would be a valuable alternative to the BEVS for rapid, scalable, and highyielding recombinant protein production and for the generation of stable Sf-9 cell lines [4]. We present a simple, efficient and cost-effective plasmidbased method for transient expression of recombinant proteins in Sf-9 cells cultivated in serum-free suspension mode in a high-throughput culture system, TubeSpin® bioreactor 50 tubes (TubeSpins)

Materials and methods

Results

Conclusions