Automation of large scale transient protein expression in mammalian cells

Automation of large scale transient protein expression in mammalian cells

Traditional mammalian expression systems rely on the time-consuming generation of stable cell lines; this is difficult to accommodate within a modern structural biology pipeline. Transient transfections are a fast, cost-effective solution, but require skilled cell culture scientists, making man-power a limiting factor in a setting where numerous samples are processed in parallel. Here we report...

Expression of Recombinant Factor IX Using the Transient Gene Expression Technique

Background: Pilot and large-scale production of recombinant proteins requires the presence of stable clones capable of producing large quantities of recombinant proteins. Not only the process of selecting stable clones is time consuming, but also the continuous culturing of clones in large-scale production may cause loss of incoming plasmid and recombinant genes. Thus, considering the advanceme...

PepGMV Rep-Protein Expression in Mammalian Cells

The Geminiviruses genome is a small, single strand DNA that replicates in the plant cell nucleus. Analogous to animal DNA viruses, Geminiviruses depend on the host replication machinery to amplify their genomes and only supply the factors required to initiate their replication. Consequently, Geminiviruses remove the cell-cycle arrest and induce the host replication machinery using an endocycle ...

Expression of Biologically Active Recombinant B-Domain-Deleted Human Factor VIII in Mammalian Cells

Serum-free large-scale transient transfection of CHO cells.

To date, methods for large-scale transient gene expression (TGE) in cultivated mammalian cells have focused on two transfection vehicles: polyethylenimine (PEI) and calcium phosphate (CaPi). Both have been shown to result in high transfection efficiencies at scales beyond 10 L. Unfortunately, both approaches yield higher levels of recombinant protein (r-protein) in the presence of serum than in...