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2021-6-19 1:27:40

Zak AJ, Hill BD, Rizvi SM, Smith MR, Yang M, Wen F. Enhancing the Yield and Quality of Influenza Virus-like Particles (VLPs) Produced in Insect Cells by Inhibiting Cytopathic Effects of Matrix Protein M2. ACS Synth Biol. 2019 Sep 5.
submited by kickingbird at Sep, 9, 2019 10:1 AM from ACS Synth Biol. 2019 Sep 5.

To provide broader protection and eliminate the need for annual update of influenza vaccines, biomolecular engineering of influenza virus-like particles (VLPs) to display more conserved influenza proteins such as the matrix protein M2 has been explored. However, achieving high surface density of full-length M2 in influenza VLPs has been left unrealized. In this study, we show that the ion channel activity of M2 induces significant cytopathic effects in Spodoptera frugiperda (Sf9) insect cells when expressed using M2-encoding baculovirus. These effects include altered Sf9 cell morphology and reduced baculovirus replication, resulting in impaired influenza protein expression and thus VLP production. Based on the function of M2, we hypothesized that blocking its ion channel activity could potentially relieve these cytopathic effects, and thus restore influenza protein expression to improve VLP production. The use of the M2 inhibitor amantadine indeed improves Sf9 cellular expression not only of M2 (~3-fold), but also of hemagglutinin (HA) (~7-fold) and of matrix protein M1 (~3-fold) when coexpressed to produce influenza VLPs. This increased cellular expression of all three influenza proteins further leads to ~2-fold greater VLP yield. More importantly, the quality of the resulting influenza VLPs is significantly improved, as demonstrated by the ~2-fold, ~50-fold, and ~2-fold increase in the antigen density to approximately 53 HA, 48 M1 and 156 M2 per influenza VLP, respectively. Taken together, this study represents a novel approach to enable the efficient incorporation of full-length M2 while enhancing both the yield and quality of influenza VLPs produced by Sf9 cells.

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