Abstract
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 (ΔPh = 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.
Highlights
Influenza is a major burden to global healthcare systems, often causing pandemics with massive death-tolls [1,2]
Increased Production of Influenza HA-Vlps in Adapted Cells
Aiming at improving the production yields of influenza HA-virus-like particles (VLPs), High Five insect cells were adapted to grow at a neutral culture pH (7.0) instead of a standard culture pH (6.2)
Summary
Influenza is a major burden to global healthcare systems, often causing pandemics with massive death-tolls [1,2]. Prophylactic vaccination still remains the most efficient approach to reduce influenza disease [3]. Due to the high tendency of influenza viruses to accumulate 3–4 amino acid substitutions per year and to genetically reassort amongst viruses in animal reservoirs [4], vaccine composition needs to be updated and new vaccines need to be formulated, produced and administrated on an annual basis, if not more often in the case of pandemics. The development of fast, economical and flexible vaccine production platforms to quickly cope with the need for high quantities of vaccine shots has become a worldwide key healthcare priority. Vaccine manufacturing platforms based on cell culture allow superior process control and flexibility, potentially improving the responsiveness to influenza epidemics or pandemics [5]. Mammalian cells have been extensively explored and proven efficient for the production of influenza vaccines [6,7]
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