Continuous high-titer HIV-1 vector production

Yasuhiro Ikeda, Yasuhiro Takeuchi, Francisco Martin, Francois Loic Cosset, Kyriacos Mitrophanous, Mary Collins

Research output: Contribution to journalArticlepeer-review

131 Scopus citations

Abstract

Human immunodeficiency virus type 1 (HIV-1)-based vectors are currently made by transient transfection, or using packaging cell lines in which expression of HIV-1 Gag and Pol proteins is induced. Continuous vector production by cells in which HIV-1 Gag-Pol is stably expressed would allow rapid and reproducible generation of large vector batches. However, attempts to make stable HIV-1 packaging cells by transfection of plasmids encoding HIV-1 Gag-Pol have resulted in cells which secrete only low levels of p24 antigen (20-80 ng/ml), possibly because of the cytotoxicity of HIV-1 protease. Infection of cells with HIV-1 can result in stable virus production; cell clones that produce up to 1,000 ng/ml secreted p24 antigen have been described. Here we report that expression of HIV-1 Gag-Pol by a murine leukemia virus (MLV) vector allows constitutive, long-term, high-level (up to 850 ng/ml p24) expression of HIV-1 Gag. Stable packaging cells were constructed using codon-optimized HIV-1 Gag-Pol and envelope proteins of gammaretroviruses; these producer cells could make up to 10 293T infectious units (i.u.)/ml (20 293T i.u./cell/day) for at least three months in culture.

Original languageEnglish (US)
Pages (from-to)569-572
Number of pages4
JournalNature biotechnology
Volume21
Issue number5
DOIs
StatePublished - May 1 2003

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering

Fingerprint Dive into the research topics of 'Continuous high-titer HIV-1 vector production'. Together they form a unique fingerprint.

Cite this