Efficient large volume lentiviral vector production using flow electroporation

Scott R. Witting, Lin Hong Li, Aparna Jasti, Cornell Allen, Kenneth Cornetta, James Brady, Rama Shivakumar, Madhusudan V. Peshwa

Research output: Contribution to journalArticle

22 Scopus citations

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 1×108 infectious units/mL. Therefore, an excellent option for implementing large-scale, clinical lentiviral productions is flow EP of suspension cell lines.

Original languageEnglish (US)
Pages (from-to)243-249
Number of pages7
JournalHuman gene therapy
Volume23
Issue number2
DOIs
StatePublished - Feb 1 2012

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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    Witting, S. R., Li, L. H., Jasti, A., Allen, C., Cornetta, K., Brady, J., Shivakumar, R., & Peshwa, M. V. (2012). Efficient large volume lentiviral vector production using flow electroporation. Human gene therapy, 23(2), 243-249. https://doi.org/10.1089/hum.2011.088