PEI-mediated transient transfection of high five cells at bioreactor scale for HIV-1 VLP production

Eduard Puente-Massaguer, Florian Strobl, Reingard Grabherr, Gerald Striedner, Martí Lecina, Francesc Gòdia

Research output: Indexed journal article Articlepeer-review

13 Citations (Scopus)


High Five cells are an excellent host for the production of virus-like particles (VLPs) with the baculovirus expression vector system (BEVS). However, the concurrent production of high titers of baculovirus hinder the purification of these nanoparticles due to similarities in their physicochemical properties. In this study, first a transient gene expression (TGE) method based on the transfection reagent polyethylenimine (PEI) is optimized for the production of HIV-1 VLPs at shake flask level. Furthermore, VLP production by TGE in High Five cells is successfully demonstrated at bioreactor scale, resulting in a higher maximum viable cell concentration (5.1 × 106 cell/mL), the same transfection efficiency and a 1.8-fold increase in Gag-eGFP VLP production compared to shake flasks. Metabolism analysis of High Five cells indicates a reduction in the consumption of the main metabolites with respect to non-transfected cell cultures, and an increase in the uptake rate of several amino acids when asparagine is depleted. Quality assessment by nanoparticle tracking analysis and flow virometry of the VLPs produced shows an average size of 100–200 nm, in agreement with immature HIV-1 viruses reported in the literature. Overall, this work demonstrates that the High Five/TGE system is a suitable approach for the production of VLP-based vaccine candidates and other recombinant proteins.

Original languageEnglish
Article number1580
Pages (from-to)1-16
Number of pages16
Issue number8
Publication statusPublished - Aug 2020


  • Bioreactor
  • High Five cells
  • Polyethylenimine
  • Transient gene expression
  • Virus-like particle


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