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Catalase effect on cell death for the improvement of recombinant protein production in baculovirus-insect cell system

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Abstract

Baculovirus expression vector system (BEVS) in host insect cells is a powerful technology to produce recombinant proteins, as well as virus-like particles (VLP). However, BEVS is based on baculovirus infection, which limits the recombinant protein production by inducing insect cell death. Herein a new strategy to enhance cell life span and to increase recombinant protein production was developed. As baculovirus infection induces cellular oxidative stress, the ability of several antioxidants to inhibit cell death was tested during infection. The production of rotavirus structural proteins was used as model to analyse this new strategy. We found that only catalase is able to partially prevent cell death triggered by baculovirus infection and to inhibit lipid peroxidation. An increase in recombinant protein production was coupled with the partial cell death inhibition. In summary, the addition of catalase is a promising strategy to improve recombinant protein production in BEVS, by delaying insect cell death.

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Abbreviations

Cat:

Catalase

Sf-9:

Spodoptera frugiperda

BEVS:

Baculovirus expression vector system

DiOC6(3):

3,3′Dihexyloxacarbocyanine iodide

ΔΨm :

Mitochondrial transmembrane potential

MMP:

Mitochondrial membrane permeabilization

PI:

Propidium iodine

hpi:

Hours post-infection

VLP:

Virus-like particles

MOI:

Multiplitity of infection

CCI:

Cell concentration at infection

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Acknowledgments

This work was supported by European Commission (QLRT200101249), the Portuguese Fundação para a Ciência e Tecnologia (POCTI/BIO/55975/2004) and the Portuguese Fundação para a Ciência e Tecnologia, for HLAV’s SFRH/BPD/14575/2003 fellowship.

The authors express their gratitude to Prof. Polly Roy from the London School of Hygiene and Tropical Medicine, England, for providing the multigene baculovirus; to Dr. J. Cohen, Dr. D. Poncet and Dr. A. Charpilienne from CNRS-INRA, France, for providing the polyclonal anti-Rotavirus serum and the baculovirus coding for VP6; and to Dr. Matthias Haury from Instituto Gulbenkian para a Ciência, Portugal, for the help with the flow cytometry measurements.

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Correspondence to Paula M. Alves.

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Vieira, H.L.A., Pereira, A.C.P., Carrondo, M.J.T. et al. Catalase effect on cell death for the improvement of recombinant protein production in baculovirus-insect cell system. Bioprocess Biosyst Eng 29, 409–414 (2006). https://doi.org/10.1007/s00449-006-0093-9

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Keywords

  • Oxidative stress
  • Catalase
  • Sf-9 insect cells
  • Cell death
  • Baculovirus
  • Rotavirus-like particles