Baculovirus-mediated production of HDV-like particles in BHK cells using a novel oscillating bioreactor

Baculovirus‐mediated gene transfer is attenuated by sodium bicarbonate

“…To enhance the gene delivery efficiency, we have previously developed a protocol by which virus transduction is performed by incubating the cells with unconcentrated virus supernatant for 4-8 h at 25 • C or 27 • C using Ca 2+ -and Mg 2+ -free Dulbecco's PBS as the surrounding solution to adjust the final liquid volume [16][17][18]. This protocol has been employed to effectively enhance baculovirusmediated gene delivery into HepG2, HeLa and BHK cells [12,19,20].…”

Generation of lentivirus vectors using recombinant baculoviruses

“…32 Baculoviruses have been widely used for a large-scale protein production in insect cells 21 and for the production of VLP, such as hepatitis VLP. 27 Intact viruses have been also produced using hybrid baculoviruses. Production of adeno-associated viruses (AAV) vector in insect cells by baculoviruses has gained much interest due to difficulties to produce enough AAV for clinical trials by conventional plasmid transfection methods.…”

“…21 Baculoviruses are capable of transducing mammalian cells 22 and have been used previously for the production of several viruses [23][24][25][26] and viral-like particles (VLP). 27 Thus, methods for the culturing and handling of baculoviruses are well developed. In order to ease and improve the lentivirus production, we constructed four recombinant baculoviruses which encode all elements needed for the lentiviral vector generation in mammalian cells.…”

Baculoviral Vectors for Gene Delivery: A Review

“…This protocol eliminates the need for virus ultracentrifugation, reduces possible virus inactivation during ultracentrifugation and represents a simpler approach. This protocol ameliorates AcMNPV-mediated gene delivery into BHK [33,51,52], Vero [48], canine MDCK [53] and chicken Df-1 cells [53], as well as augments the transduction of HEK293 and Schwann cells mediated by another baculovirus Bombyx mori NPV (BmNPV) [54]. One key determinant accounting for the improved transduction is the surrounding solution.…”

“…Cotransduction of hepatoma cell lines with these two baculoviruses leads to the secretion of HDV VLPs with the yield approximately 2 times that achieved by plasmid transfection [33]. The VLPs production process is further switched to the FDA-approved BHK cells and scaled up in a novel oscillating packed-bed bioreactor [51]. Similar to HDV, HCV does not replicate in cultured cells, but HCV VLP production is recently achieved by transducing human hepatoma cell lines (e.g.…”