Kinetic analysis of in vitro production of wild-type Spodoptera frugiperda nucleopolyhedrovirus

Almeida, Andréa Farias de; Macêdo, Gorete Ribeiro de; Chan, Leslie C. L.; Pedrini, Márcia Regina da Silva

doi:10.1590/s1516-89132010000200006

Cited by 13 publications

(12 citation statements)

References 6 publications

(8 reference statements)

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“…Studies on Group I CfMNPV in CF‐2C1 cells57 and BmNPV in Bm5 cells55 did not follow BV production, but the modest cell growth reported after application of a low MOI (0.1 PFU per cell) was similar to that reported for AcMNPV at the same MOI,5 which suggested similarly fast infection kinetics. Studies on Group II SfMNPV in Sf9 cells58 also did not monitor BV production, but the infection kinetics appeared to be poor because the application of high MOIs (by adding 10% v/v of virus inoculum) still resulted in a doubling in cell density postinfection, similar to the experience with HaSNPV 20, 48…”

Section: Discussionmentioning

confidence: 88%

Kinetic characterization of the group II helicoverpa armigera nucleopolyhedrovirus propagated in suspension cell cultures: Implications for development of a biopesticides production process

Pedrini

Reid

Nielsen

et al. 2011

Biotechnology Progress

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Large-scale commercialization of baculovirus biopesticides for the control of insect pests requires a cell culture production process, and knowledge of the infection kinetics is a vital prerequisite for process optimization. Well-characterized kinetic parameters have so far only been reported for the commercially established recombinant Autographa californica nucleopolyhedrovirus (AcMNPV), a Group I NPV. In this work, key infection kinetic parameters of the Group II NPV Helicoverpa armigera nucleopolyhedrovirus (HaSNPV), and its Few Polyhedra (FP) mutant, were well characterized for the first time, in suspension HzAM1 insect cell cultures, to facilitate the scale-up of an HaSNPV-based biopesticide. The FP mutant had a selective advantage over wild-type HaSNPV in cell cultures, and the kinetic analysis showed that this was due to a superior budding rate, rather than a faster binding rate (BR) or longer budding duration. Another finding was that wild-type HaSNPV had very poor infection kinetics when compared with AcMNPV, exhibiting an 18-fold lower BR, a more than 50-fold lower budding rate, and a 60-fold lower extracellular/total progeny virus ratio. Such poor infection kinetics have serious implications during scale-up of an HaSNPV biopesticide production process, including the requirement for large volumes of virus inocula and the difficulty of achieving synchronous infections. Groups I and II NPVs may have very different infection kinetics because of their different envelope fusion proteins. This study is the first to compare the two groups of NPVs in terms of well-characterized cell-specific infection kinetics, and the findings may indicate a phylogenetic basis for kinetic differences.

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Section: Discussionmentioning

confidence: 88%

Kinetic characterization of the group II helicoverpa armigera nucleopolyhedrovirus propagated in suspension cell cultures: Implications for development of a biopesticides production process

Pedrini

Reid

Nielsen

et al. 2011

Biotechnology Progress

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“…Within the first 8 days, the inoculum of 5 × 10 7 cells had expanded to a maximum total cell count of 2. ) [54][55][56]. For the evaluation of infected cells, the same setup was used with the supplementation of 1% infected cells in the initial inoculum ( Figure 5).…”

Section: Bves In Shear-free Environmentmentioning

confidence: 99%

The components of shear stress affecting insect cells used with the baculovirus expression vector system

Weidner

Druzinec

Mühlmann

et al. 2017

Zeitschrift Für Naturforschung C

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Insect-based expression platforms such as the baculovirus expression vector system (BEVS) are widely used for the laboratory-and industrial-scale production of recombinant proteins. Thereby, major drawbacks to gain high-quality proteins are the lytic infection cycle and the shear sensitivity of infected insect cells due to turbulence and aeration. Smaller bubbles were formerly assumed to be more harmful than larger ones, but we found that cell damage is also dependent on the concentration of protective agents such as Pluronic ® . At the appropriate concentration, Pluronic forms a layer around air bubbles and hinders the attachment of cells, thus limiting the damage. In this context, we used microaeration to vary bubble sizes and confirmed that size is not the most important factor, but the total gas surface area in the reactor is. If the surface area exceeds a certain threshold, the concentration of Pluronic is no longer sufficient for cell protection. To investigate the significance of shear forces, a second study was carried out in which infected insect cells were cultivated in a hollow fiber module to protect them from shear forces. Both model studies revealed important aspects of the design and scale-up of BEVS processes for the production of recombinant proteins.

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“…Cell cultures of insects are an interesting means to select systems that support viral replication (Castro et al, 2006;Almeida et al, 2010;Huynh et al, 2015). These cell systems are useful in studies of virus-host interactions, but the present technical limitations should be solved for their need in large-scale production of biopesticides, such as the accumulation of "few polyhedra mutants" or "defective interfering particles" in the cell culture (Moscardi et al, 2011;Reid et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

“…Selecting productive cell lines is an essential initial step for the establishment of SfMNPV in vitro production. Previous work with other SfMNPV isolate 18, which is less virulent than the isolate 19, was done exclusively with S. frugiperda cell lines in suspension culture (Almeida et al, 2010). However, it is well known that baculovirus species can infect cell lines originated from other Lepidoptera species.…”

Section: Introductionmentioning

confidence: 99%

In vitro infectivity of Spodoptera frugiperda multiple nucleopolyhedrovirus to different insect cell lines

Sihler

Souza

Valicente

et al. 2018

Pesq. agropec. bras.

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The objective of this work was to evaluate the response of an in vitro host range to Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV), a pathogenic virus to the fall armyworm (Spodoptera frugiperda, Lepidoptera: Noctuidae), for the further development of a biopesticide based on cell culture systems. The cell lines from Bombyx mori (BM-5), Lymantria dispar (IPLB-LD-625Y), Trichoplusia ni (BTI-Tn-5B1-4), Anticarsia gemmatalis (UFL-AG-286), and S. frugiperda (IPLB-SF-21AE and Sf9) were tested for their susceptibility to a highly-virulent Brazilian isolate of SfMNPV. The cytopathic effects induced by the virus, the production of viral particles, and the synthesis of viral polypeptides were examined and compared. Both S. frugiperda cell lines showed hypertrophy of cell nuclei and production of many polyhedra. The SDS-Page of radiolabed proteins showed that the cell protein synthesis was shutoff, while an intense band of about 30 kDa, recognized as polyhedrin, was synthesized. The other cell lines did not show polyhedra production, although some of them underwent morphological changes and protein synthesis shutdown in response to virus infection. The SF-21 and Sf9 cell lines are recommended for further in vitro production of SfMNPV.

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Kinetic analysis of in vitro production of wild-type Spodoptera frugiperda nucleopolyhedrovirus

Cited by 13 publications

References 6 publications

Kinetic characterization of the group II helicoverpa armigera nucleopolyhedrovirus propagated in suspension cell cultures: Implications for development of a biopesticides production process

Kinetic characterization of the group II helicoverpa armigera nucleopolyhedrovirus propagated in suspension cell cultures: Implications for development of a biopesticides production process

The components of shear stress affecting insect cells used with the baculovirus expression vector system

In vitro infectivity of Spodoptera frugiperda multiple nucleopolyhedrovirus to different insect cell lines

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