title: Decline in Helicoverpa armigera nucleopolyhedrovirus occlusion body yields with increasing infection cell density in vitro is strongly correlated with viral DNA levels.
abstract: The phenomenon of the reduction in the cell-specific yield with increasing infection cell density (ICD), the cell density effect, is one of the main hurdles for improving virus yields in vitro. In the current study, the reduction in cell-specific yields (viral DNA [vDNA], polyhedrin mRNA and occlusion body [OB]) with increasing ICD for Helicoverpa armigera nucleopolyhedrovirus (HearNPV)-infected HzAM1 (Helicoverpa zea) insect cells has been investigated. HzAM1 cells were propagated in Sf900™ III serum-free medium and synchronously infected with wild-type HearNPV at various ICDs of 0.5-5 × 10(6) cells/mL at an MOI of 5 PFU/cell. Infection was conducted either in the original medium or in fresh medium. As found previously for Sf9 and High Five cells, there were negative correlations between the three key virus infection indicators (vDNA, mRNA and OB) and the peak cell density (PCD). Generally, the yield decline with increasing PCD was most pronounced for OB, followed by mRNA, and was more moderate for vDNA. The decline was significantly reduced, but not totally arrested, when fresh medium was used. There were also strong correlations between OB and mRNA, mRNA and vDNA, and OB and vDNA levels. These results suggest that the reduction in baculovirus yield (OB) at high PCDs is associated with limitations during the upstream processes of replication and transcription together with limitations during protein translation. Furthermore, the peak protein productivity per unit of cell volume in the HzAM1/HearNPV system was shown to be higher than that of the Sf9/rAcMNPV system, but lower than that of the High Five/rAcMNPV system.
authors.affiliation: Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St Lucia, QLD, 4072, Australia, hoai.huynh@uqconnect.edu.au.
authors.forename: Hoai T
authors.initials: HT
authors.lastname: Huynh
authors.forename: Trinh T B
authors.initials: TT
authors.lastname: Tran
authors.forename: Leslie C L
authors.initials: LC
authors.lastname: Chan
authors.forename: Lars K
authors.initials: LK
authors.lastname: Nielsen
authors.forename: Steven
authors.initials: S
authors.lastname: Reid
mesh_terms: D000818:Animals; D002452:Cell Count; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry; D004279:DNA, Viral / Q000032:analysis*; D007181:Inclusion Bodies, Viral; D017924:Nucleopolyhedroviruses / Q000254:growth & development*; D012333:RNA, Messenger / Q000032:analysis; D018411:Spodoptera; D014776:Virus Cultivation
chemical_list: D003470:Culture Media; D004279:DNA, Viral; D012333:RNA, Messenger