Enhancing the Yield and Quality of Influenza Virus-like
Particles (VLPs) Produced in Insect Cells by Inhibiting Cytopathic
Effects of Matrix Protein M2
Posted on 2019-09-13 - 12:33
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. On the basis of 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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Zak, Andrew
J.; Hill, Brett D.; Rizvi, Syed M.; Smith, Mason R.; Yang, Madeleine; Wen, Fei (2019). Enhancing the Yield and Quality of Influenza Virus-like
Particles (VLPs) Produced in Insect Cells by Inhibiting Cytopathic
Effects of Matrix Protein M2. ACS Publications. Collection. https://doi.org/10.1021/acssynbio.9b00111
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AUTHORS (6)
AZ
Andrew
J. Zak
BH
Brett D. Hill
SR
Syed M. Rizvi
MS
Mason R. Smith
MY
Madeleine Yang
FW
Fei Wen
KEYWORDS
Sf 9 cell morphology156 M 2Inhibiting Cytopathic Effectsinfluenza virus-like particlesM 2-encoding baculovirusion channel activityinfluenza proteinsHAMatrix Protein M 2influenza protein expressionSf 9 cellsinfluenza VLPsVLP productionM 2M 2 inhibitor amantadinematrix protein M 2Influenza Virus-like Particlescytopathic effects