The method used to culture host cells (Sf9 cells) can affect the qualities of baculovirus budding particles expressing recombinant proteins.
Budded virus (BV) particles of baculovirus (Autographa californica nucleopolyhedrovirus, AcNPV) are harvested from the supernatant of liquid culture of Sf9 host cells by ultracentrifugation. Using polyacrylamide gel electrophoresis, Western blot and transmission electron microscopy (TEM) of BV samples fractionated closely by sucrose density gradient centrifugation, we observed that BVs exhibited different qualities depending on whether they had been harvested from the supernatant from a standing (static), shaking (suspension), or standing/shaking (pre-/post-infection) culture of Sf9 cells. The amount of BV protein apparently increased in the order of standing, standing/shaking, and shaking procedure, and the yield of intact particles showed an opposite trend. TEM observation clearly showed that appropriate fractions of the standing and standing/shaking cultures contained more intact BV particles than those from the shaking culture. These results suggest that the qualities of recombinant BV particles may be related to the culture conditions of the host cells.
a Division of Chemistry for Materials , Graduate School of Engineering, Mie University , Tsu , Japan.
Tomomi
T
Hattori
a Division of Chemistry for Materials , Graduate School of Engineering, Mie University , Tsu , Japan.
Kohei
K
Nakanishi
a Division of Chemistry for Materials , Graduate School of Engineering, Mie University , Tsu , Japan.
Takaaki
T
Mori
a Division of Chemistry for Materials , Graduate School of Engineering, Mie University , Tsu , Japan.
Masahiro
M
Tomita
a Division of Chemistry for Materials , Graduate School of Engineering, Mie University , Tsu , Japan.
Kanta
K
Tsumoto
D000818:Animals; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005814:Genes, Viral; D046529:Microscopy, Electron, Transmission; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics; D061987:Sf9 Cells; D018411:Spodoptera; D014461:Ultracentrifugation
D011994:Recombinant Proteins
baculovirus; budded virus; electron microscopy; recombinant protein; sucrose density gradient