Accelerating HIV-1 VLP production using stable High Five insect cell pools

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

Research output: Indexed journal article Articlepeer-review

13 Citations (Scopus)

Abstract

Stable cell pools are receiving a renewed interest as a potential alternative system to clonal cell lines. The shorter development timelines and the capacity to achieve high product yields make them an interesting approach for recombinant protein production. In this study, stable High Five cell pools are assessed for the production of a simple protein, mCherry, and the more complex HIV-1 Gag-eGFP virus-like particles (VLPs). Random integration coupled to fluorescence-activated cell sorting (FACS) in suspension conditions is applied to accelerate the stable cell pool generation process and enrich it with high producer cells. This methodology is successfully transferred to a bioreactor for VLP production, resulting in a 2-fold increase in VLP yields with respect to shake flask cultures. In these conditions, maximum viable cell concentration improves by 1.5-fold, and by-product formation is significantly reduced. Remarkably, a global increase in the uptake of amino acids in the Gag-eGFP stable cell pool is observed when compared with parental High Five cells, reflecting the additional metabolic burden associated with VLP production. These results suggest that stable High Five cell pools are a robust and powerful approach to produce VLPs and other recombinant proteins, and put the basis for future studies aiming to scale up this system.

Original languageEnglish
Article number2000391
JournalBiotechnology Journal
Volume16
Issue number4
DOIs
Publication statusPublished - Apr 2021

Keywords

  • High Five cells
  • bioreactor
  • fluorescence-activated cell sorting
  • metabolism
  • stable cell pool
  • virus-like particle

Fingerprint

Dive into the research topics of 'Accelerating HIV-1 VLP production using stable High Five insect cell pools'. Together they form a unique fingerprint.

Cite this