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Efficient large volume lentiviral vector production using flow electroporation.

ABSTRACT: 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 1×10(8) infectious units/mL. Therefore, an excellent option for implementing large-scale, clinical lentiviral productions is flow EP of suspension cell lines.

SUBMITTER: Witting SR

PROVIDER: S-EPMC3311912 | biostudies-literature | 2012 Feb

REPOSITORIES: biostudies-literature

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Efficient large volume lentiviral vector production using flow electroporation.

Witting Scott R SR Li Lin-Hong LH Jasti Aparna A Allen Cornell C Cornetta Kenneth K Brady James J Shivakumar Rama R Peshwa Madhusudan V MV

Human gene therapy 20111024 2

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 g ...[more]

PMID: 21933028

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