Next Article in Journal
Resin-Based Sealant with Bioactive Glass and Zwitterionic Material for Remineralisation and Multi-Species Biofilm Inhibition
Next Article in Special Issue
Sono-Assembly of the [Arg-Phe]4 Octapeptide into Biofunctional Nanoparticles
Previous Article in Journal
Design and Tuning of Nanofluids Applied to Chemical Enhanced Oil Recovery Based on the Surfactant–Nanoparticle–Brine Interaction: From Laboratory Experiments to Oil Field Application
Previous Article in Special Issue
ZnO Nano-Particles Production Intensification by Means of a Spinning Disk Reactor
Article

PEI-Mediated Transient Transfection of High Five Cells at Bioreactor Scale for HIV-1 VLP Production

1
Departament d’Enginyeria Química, Biològica i Ambiental, Universitat Autònoma de Barcelona, 08193 Barcelona, Spain
2
Austrian Centre of Industrial Biotechnology (acib GmbH), 1010 Vienna, Austria
3
Department of Biotechnology, University of Natural Resources and Life Sciences, 1190 Vienna, Austria
4
IQS School of Engineering, Universitat Ramón Llull, 08017 Barcelona, Spain
*
Author to whom correspondence should be addressed.
Nanomaterials 2020, 10(8), 1580; https://doi.org/10.3390/nano10081580
Received: 22 June 2020 / Revised: 20 July 2020 / Accepted: 7 August 2020 / Published: 12 August 2020
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. View Full-Text
Keywords: High Five cells; transient gene expression; polyethylenimine; virus-like particle; bioreactor High Five cells; transient gene expression; polyethylenimine; virus-like particle; bioreactor
Show Figures

Figure 1

MDPI and ACS Style

Puente-Massaguer, E.; Strobl, F.; Grabherr, R.; Striedner, G.; Lecina, M.; Gòdia, F. PEI-Mediated Transient Transfection of High Five Cells at Bioreactor Scale for HIV-1 VLP Production. Nanomaterials 2020, 10, 1580. https://doi.org/10.3390/nano10081580

AMA Style

Puente-Massaguer E, Strobl F, Grabherr R, Striedner G, Lecina M, Gòdia F. PEI-Mediated Transient Transfection of High Five Cells at Bioreactor Scale for HIV-1 VLP Production. Nanomaterials. 2020; 10(8):1580. https://doi.org/10.3390/nano10081580

Chicago/Turabian Style

Puente-Massaguer, Eduard, Florian Strobl, Reingard Grabherr, Gerald Striedner, Martí Lecina, and Francesc Gòdia. 2020. "PEI-Mediated Transient Transfection of High Five Cells at Bioreactor Scale for HIV-1 VLP Production" Nanomaterials 10, no. 8: 1580. https://doi.org/10.3390/nano10081580

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop