Coupling microscopy and flow cytometry for a comprehensive characterization of nanoparticle production in insect cells

Assess and compare the Gag-eGFP VLP production process in the Sf9 and High Five insect cell lines by AcMNPV baculovirus infection.
Conclusion:
The AcMNPV baculovirus infection process progresses faster in Sf9 cells but High Five cells display a marked increase in their complexity (SSC-H), which is not observed in the former. This increase in cell complexity as the baculovirus infection advances is attributed to the production of higher levels of occlusion-derived baculoviruses in High Five over Sf9 cells. A higher concentration of Gag-eGFP VLPs is achieved in Sf9 cells although High Five cells produce larger amounts of the Gag-eGFP polyprotein. Moreover, the interaction of High Five-derived VLPs with cellular compounds is detected, which increases the sedimentation coefficient of these nanoparticles DPBS and fresh Sf900III medium were used as controls. Furthermore, non-infected sample supernatants (0 hours post infection) and detergent lysis controls were also included. All experiments were performed in triplicate