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

19 Citations (Scopus)

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

Fingerprint

Electroporation
Transfection
Suspensions
Cell Line
Vesicular Stomatitis
Genetic Therapy
Glycoproteins
HIV
Serum

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Witting, S. R., Li, L. H., Jasti, A., Allen, C., Cornetta, K., Brady, J., ... 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

Efficient large volume lentiviral vector production using flow electroporation. / Witting, Scott R.; Li, Lin Hong; Jasti, Aparna; Allen, Cornell; Cornetta, Kenneth; Brady, James; Shivakumar, Rama; Peshwa, Madhusudan V.

In: Human gene therapy, Vol. 23, No. 2, 01.02.2012, p. 243-249.

Research output: Contribution to journalArticle

Witting, SR, Li, LH, Jasti, A, Allen, C, Cornetta, K, Brady, J, Shivakumar, R & Peshwa, MV 2012, 'Efficient large volume lentiviral vector production using flow electroporation', Human gene therapy, vol. 23, no. 2, pp. 243-249. https://doi.org/10.1089/hum.2011.088
Witting, Scott R. ; Li, Lin Hong ; Jasti, Aparna ; Allen, Cornell ; Cornetta, Kenneth ; Brady, James ; Shivakumar, Rama ; Peshwa, Madhusudan V. / Efficient large volume lentiviral vector production using flow electroporation. In: Human gene therapy. 2012 ; Vol. 23, No. 2. pp. 243-249.
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