Improving influenza HA-VLPs production in insect high five cells via adaptive laboratory evolution

Ricardo Correia, Bárbara Fernandes, Paula M. Alves, Manuel J.T. Carrondo, António Roldão

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


The use of non-standard culture conditions has proven efficient to increase cell performance and recombinant protein production in different cell hosts. However, the establishment of high-producing cell populations through adaptive laboratory evolution (ALE) has been poorly explored, in particular for insect cells. In this study, insect High Five cells were successfully adapted to grow at a neutral culture pH (7.0) through ALE for an improved production of influenza hemagglutinin (HA)-displaying virus-like particles (VLPs). A stepwise approach was used for the adaptation process, in which the culture pH gradually increased from standard 6.2 to 7.0 (∆Ph = 0.2–0.3), and cells were maintained at each pH value for 2–3 weeks until a constant growth rate and a cell viability over 95% were observed. These adapted cells enabled an increase in cell-specific HA productivity up to three-fold and volumetric HA titer of up to four-fold as compared to non-adapted cells. Of note, the adaptation process is the element driving increased specific HA productivity as a pH shift alone was inefficient at improving productivities. The production of HA-VLPs in adapted cells was successfully demonstrated at the bioreactor scale. The produced HA-VLPs show the typical size and morphology of influenza VLPs, thus confirming the null impact of the adaptation process and neutral culture pH on the quality of HA-VLPs produced. This work strengthens the potential of ALE as a bioprocess engineering strategy to improve the production of influenza HA-VLPs in insect High Five cells.

Original languageEnglish
Article number589
Pages (from-to)1-15
Number of pages15
Issue number4
Publication statusPublished - Dec 2020


  • Adaptive laboratory evolution
  • Baculovirus expression system
  • Improved production
  • Influenza HA-VLPs
  • Insect High Five cells


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