Quantitative proteomic analysis of lentiviral vectors using 2-DE

Jérôme Denard, Stéphanie Rundwasser, Nicolas Laroudie, Florence Gonnet, Luigi Naldini, Marina Radrizzani, Anne Galy, Otto Wilhelm Merten, Olivier Danos, Fedor Svinartchouk

Research output: Contribution to journalArticle

Abstract

Among the integrative gene therapy vectors developed to date, human immunodeficiency virus type 1 (HIV-1)-derived lentiviral vectors (LV) are distinguished by their capacity to infect both dividing and non-dividing cells. Recombinant LV particles contain viral proteins necessary for their packaging, infectious and integrating functions. Like the parental HIV-1 virus they are able to acquire various cellular proteins, but the number and localisation of these proteins are poorly characterised. In the present study we used 2-DE followed by MALDI-TOF to quantify the protein content of several types of vesicular stomatitis virus G-pseudotyped LV including those that were extensively purified in the perspective of clinical gene therapy studies. A proteinase K treatment was used to distinguish between cellular proteins incorporated into virions (I-proteins) and those co-purified with vectors (C-proteins). We found 10 C-proteins and 18 I-proteins associated with LV. Copy numbers for these core I-proteins varied from 5 (AIP-1/ALIX) to 280 (Cyclophilin A) per vector particle. Three novel I-proteins, guanine nucleotide-binding protein 2, L-lactate dehydrogenase B chain and hnRNP core protein A1, were found. This study defines for the first time, the protein stoichiometry of infectious HIV-1-derived LV particles.

Original languageEnglish
Pages (from-to)3666-3676
Number of pages11
JournalProteomics
Volume9
Issue number14
DOIs
Publication statusPublished - Jul 2009

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Keywords

  • 2-DE
  • Cellular proteins
  • Gene therapy
  • Lentiviral vector

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

Cite this

Denard, J., Rundwasser, S., Laroudie, N., Gonnet, F., Naldini, L., Radrizzani, M., Galy, A., Merten, O. W., Danos, O., & Svinartchouk, F. (2009). Quantitative proteomic analysis of lentiviral vectors using 2-DE. Proteomics, 9(14), 3666-3676. https://doi.org/10.1002/pmic.200800747