An Alternative Perfusion Approach for the Intensification of Virus-Like Particle Production in HEK293 Cultures
Lavado-García, Jesús (Universitat Autònoma de Barcelona. Escola d'Enginyeria)
Cervera Gracia, Laura (Universitat Autònoma de Barcelona. Escola d'Enginyeria)
Gòdia, Francesc (Universitat Autònoma de Barcelona. Escola d'Enginyeria)

Fecha:	2020
Resumen:	Virus-like particles (VLPs) have gained interest over the last years as recombinant vaccine formats, as they generate a strong immune response and present storage and distribution advantages compared to conventional vaccines. Therefore, VLPs are being regarded as potential vaccine candidates for several diseases. One requirement for their further clinical testing is the development of scalable processes and production platforms for cell-based viral particles. In this work, the extended gene expression (EGE) method, which consists in consecutive media replacements combined with cell retransfections, was successfully optimized and transferred to a bioreactor operating in perfusion. A process optimization using design of experiments (DoE) was carried out to obtain optimal values for the time of retransfection, the cell specific perfusion rate (CSPR) and transfected DNA concentration, improving 86. 7% the previously reported EGE protocol in HEK293. Moreover, it was successfully implemented at 1. 5L bioreactor using an ATF as cell retention system achieving concentrations of 6. 8·10 10 VLP/mL. VLP interaction with the ATF hollow fibers was studied via confocal microscopy, field emission scanning electron microscopy, and nanoparticle tracking analysis to design a bioprocess capable of separating unassembled Gag monomers and concentrate VLPs in one step.
Ayudas:	Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-898
Nota:	Altres ajuts: Fundació la Caixa (LCF/BQ/ES17/11600003)
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Bioreactor ; Perfusion ; ATF ; Design of experiments ; VLP ; HFM
Publicado en:	Frontiers in Bioengineering and Biotechnology, Vol. 8 (June 2020) , art. 617, ISSN 2296-4185

DOI: 10.3389/fbioe.2020.00617
PMID: 32637402

16 p, 3.9 MB

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