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

23 Citations (Scopus)

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

Fingerprint

Proteomics
Viruses
Proteins
HIV-1
Gene therapy
Protein C
Genetic Therapy
Cyclophilin A
Endopeptidase K
Heterogeneous-Nuclear Ribonucleoproteins
Vesicular Stomatitis
Guanine Nucleotides
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Viral Proteins
Product Packaging
L-Lactate Dehydrogenase
Virion
Stoichiometry
Carrier Proteins
Packaging

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., ... Svinartchouk, F. (2009). Quantitative proteomic analysis of lentiviral vectors using 2-DE. Proteomics, 9(14), 3666-3676. https://doi.org/10.1002/pmic.200800747

Quantitative proteomic analysis of lentiviral vectors using 2-DE. / Denard, Jérôme; Rundwasser, Stéphanie; Laroudie, Nicolas; Gonnet, Florence; Naldini, Luigi; Radrizzani, Marina; Galy, Anne; Merten, Otto Wilhelm; Danos, Olivier; Svinartchouk, Fedor.

In: Proteomics, Vol. 9, No. 14, 07.2009, p. 3666-3676.

Research output: Contribution to journalArticle

Denard, J, Rundwasser, S, Laroudie, N, Gonnet, F, Naldini, L, Radrizzani, M, Galy, A, Merten, OW, Danos, O & Svinartchouk, F 2009, 'Quantitative proteomic analysis of lentiviral vectors using 2-DE', Proteomics, vol. 9, no. 14, pp. 3666-3676. https://doi.org/10.1002/pmic.200800747
Denard J, Rundwasser S, Laroudie N, Gonnet F, Naldini L, Radrizzani M et al. Quantitative proteomic analysis of lentiviral vectors using 2-DE. Proteomics. 2009 Jul;9(14):3666-3676. https://doi.org/10.1002/pmic.200800747
Denard, Jérôme ; Rundwasser, Stéphanie ; Laroudie, Nicolas ; Gonnet, Florence ; Naldini, Luigi ; Radrizzani, Marina ; Galy, Anne ; Merten, Otto Wilhelm ; Danos, Olivier ; Svinartchouk, Fedor. / Quantitative proteomic analysis of lentiviral vectors using 2-DE. In: Proteomics. 2009 ; Vol. 9, No. 14. pp. 3666-3676.
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