Efficient Large Volume Lentiviral Vector Production Using Flow Electroporation

Lentiviral vectors are beginning to emerge as a viable choice for human gene therapy. Here, we describe a method that combines the convenience of a suspension cell line with a scalable, nonchemically based, and GMP-compliant transfection technique known as flow electroporation (EP). Flow EP parameters for serum-free adapted HEK293FT cells were optimized to limit toxicity and maximize titers. Using a third generation, HIV-based, lentiviral vector system pseudotyped with the vesicular stomatitis glycoprotein envelope, both small- and large-volume transfections produced titers over 1×108 infectious units/mL. Therefore, an excellent option for implementing large-scale, clinical lentiviral productions is flow EP of suspension cell lines. Witting and colleagues describe a flow electroporation (flow EP) transfection method for efficient, scalable, nonchemically based and GMP-compliant production of lentiviral vectors. Both small- and large-scale flow EP transfections of serum-free adapted HEK293FT cells using a third-generation HIV-based lentiviral vector system produce titers over 1 × 108 infectious units/ml.