title: Characterization of an Sf-rhabdovirus-negative Spodoptera frugiperda cell line as an alternative host for recombinant protein production in the baculovirus-insect cell system.
abstract: Cell lines derived from the fall armyworm, Spodoptera frugiperda (Sf), are widely used as hosts for recombinant protein production in the baculovirus-insect cell system (BICS). However, it was recently discovered that these cell lines are contaminated with a virus, now known as Sf-rhabdovirus [1]. The detection of this adventitious agent raised a potential safety issue that could adversely impact the BICS as a commercial recombinant protein production platform. Thus, we examined the properties of Sf-RVN, an Sf-rhabdovirus-negative Sf cell line, as a potential alternative host. Nested RT-PCR assays showed Sf-RVN cells had no detectable Sf-rhabdovirus over the course of 60 passages in continuous culture. The general properties of Sf-RVN cells, including their average growth rates, diameters, morphologies, and viabilities after baculovirus infection, were virtually identical to those of Sf9 cells. Baculovirus-infected Sf-RVN and Sf9 cells produced equivalent levels of three recombinant proteins, including an intracellular prokaryotic protein and two secreted eukaryotic glycoproteins, and provided similar N-glycosylation patterns. In fact, except for the absence of Sf-rhabdovirus, the only difference between Sf-RVN and Sf9 cells was SF-RVN produced higher levels of infectious baculovirus progeny. These results show Sf-RVN cells can be used as improved, alternative hosts to circumvent the potential safety hazard associated with the use of Sf-rhabdovirus-contaminated Sf cells for recombinant protein manufacturing with the BICS.
authors.affiliation: GlycoBac, LLC, Laramie, WY 82072, USA.
authors.forename: Ajay B
authors.initials: AB
authors.lastname: Maghodia
authors.affiliation: GlycoBac, LLC, Laramie, WY 82072, USA.
authors.forename: Christoph
authors.initials: C
authors.lastname: Geisler
authors.affiliation: GlycoBac, LLC, Laramie, WY 82072, USA; Department of Molecular Biology, University of Wyoming, Laramie, WY 82071, USA. Electronic address: dljarvis@uwyo.edu.
authors.forename: Donald L
authors.initials: DL
authors.lastname: Jarvis
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods; D004921:Erythropoietin / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D009174:Mycoplasma / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000235:genetics; D012209:Rhabdoviridae / Q000302:isolation & purification*; D061987:Sf9 Cells; D018411:Spodoptera / Q000166:cytology* / Q000235:genetics / Q000821:virology*
chemical_list: C554270:EPO protein, human; D011994:Recombinant Proteins; D004921:Erythropoietin
keywords: Baculovirus; Insect cells; Recombinant protein production; Sf-RVN cells; Sf-rhabdovirus