EFFECT OF M1 MATRIX PROTEIN ON HEMAGGLUTININ EXPRESSION AND INFLUENZA VIRUS-LIKE PARTICLE (VLP) PRODUCTION IN TRICHOPLUSIA NI
Abstract
Virus-like particles (VLPs) are a new flu vaccine approach that is gaining traction because VLPs are non-infectious and can produce robust immune responses. Flu VLPs are historically created by co-expressing the flu hemagglutinin (HA) surface antigen and M1 matrix proteins in tissue culture cells. However, theoretical advantages exist for producing HA-only VLPs in living bioreactors. Reducing or eliminating M1 may result in higher HA expression and therefore more efficient vaccine production. This project examined the effect of M1 matrix protein on HA expression and VLP production in a Trichoplusia ni living bioreactor expression system. T. ni pupae were injected with varying doses and ratios of influenza HA and M1 baculoviruses. On day 3 post-infection, the pupae were extracted and a VLP-enriched particulate fraction prepared by differential centrifugation. Samples were analyzed by gel electrophoresis and Western blotting. We found that exclusion of M1 does not preclude formation of VLP-like particles, and that HA expression increases as M1 decreases. We also found that lower doses of infecting baculovirus(es) correlated with higher HA expression. Transmission electron microscopy studies are planned to investigate the effects of matrix M1 content on VLP morphology. The goal of this research is to maximize flu HA VLP expression in a T. ni pupal bioreactor expression system, and enable rapid and economical production of material for pre-clinical evaluation of broadly-protective flu vaccine candidates.
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