Abstract
Insect cells have been widely used for the production of recombinant proteins using recombinant baculovirus for gene delivery [1]. To simplify protein production in insect cells, we have previously described a method, based on transient gene expression (TGE) with cultures of suspension-adapted Sf9 cells using polyethylenimine (PEI) for DNA delivery [2]. Expression of GFP has been realized at high efficiency and a tumor necrosis factor receptor-Fc fusion protein (TNFR-Fc) was produced at a level of 40 mg/L. However, the efficiency of the insect cells TGE system has not been studied and further opti- mization may improve protein titers. Here, we studied the efficiency of PEI for plasmid delivery in Sf9 cells.
Highlights
Insect cells have been widely used for the production of recombinant proteins using recombinant baculovirus for gene delivery [1]
Plasmid delivery efficiency in Sf9 cells To measure the time course of plasmid DNA delivery, cells were transfected with a GFP expression vector
To measure the amount of DNA uptake, Sf9 cells were transfected in two different ways with a tumor necrosis factor receptor-Fc fusion protein (TNFR-Fc) expression vector and the amount of intracellular plasmid was measured by quantitative PCR
Summary
Insect cells have been widely used for the production of recombinant proteins using recombinant baculovirus for gene delivery [1]. To simplify protein production in insect cells, we have previously described a method, based on transient gene expression (TGE) with cultures of suspension-adapted Sf9 cells using polyethylenimine (PEI) for DNA delivery [2]. Expression of GFP has been realized at high efficiency and a tumor necrosis factor receptor-Fc fusion protein (TNFR-Fc) was produced at a level of 40 mg/L. The efficiency of the insect cells TGE system has not been studied and further optimization may improve protein titers. We studied the efficiency of PEI for plasmid delivery in Sf9 cells
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