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Production of biologically active feline interferon beta in insect larvae using a recombinant baculovirus

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Abstract

Feline interferon beta is a cytokine that belongs to the type I IFN family, with antitumor, antiviral and immunomodulatory functions. In this work, recombinant feline interferon beta (rFeIFNβ) was expressed in insect larvae that constitute important agronomic plagues. rFeIFNβ accumulated in the hemolymph of Spodoptera frugiperda larvae infected with recombinant baculovirus and was purified by Blue-Sepharose chromatography directly from larval homogenates on day 4 post-infection. rFeIFNβ was recovered after purification with a specific activity of 1 × 106 IU mg−1. By this method, we obtained 8.9 × 104 IU of purified rFeIFNβ per larva. The product was biologically active in vitro, with an antiviral activity of 9.5 × 104 IU mL−1, as well as a potent antitumor activity comparable to that of the commercial FeIFNω. The glycosylation of rFeIFNβ was confirmed by peptide-N-glycosidase F digestion. Our findings provide a cost-effective platform for large-scale rFeIFNβ production in laboratory research or veterinary medicine applications.

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Abbreviations

AcMNPV:

Autographa californica multiple nucleopolyhedrovirus

FeIFN:

Feline interferon

rFeIFN:

Recombinant feline interferon

MW:

Molecular weight

FBS:

Fetal bovine serum

VSV:

Vesicular stomatitis virus

MEM:

Modified Eagle’s medium

MOI:

Multiplicity of infection

DPI:

Day(s) post-infection

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Acknowledgements

This work was supported by Grants from Agencia Nacional de Promoción Científica y Tecnológica de Argentina and from Universidad de Buenos Aires (PICT 2014-3350; UBACyT 2016-20020150100145BA). AMT, MSV, FJW and MVM are career researchers of the Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET). MBA is a research fellow of CONICET. GJM is a research fellow of Agencia de Promoción Científica y Tecnológica (ANPCyT).

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Authors and Affiliations

  1. Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Universidad de Buenos Aires, Junín 956, 1113, Buenos Aires, Argentina

    Mariana Bernadett Arregui, Gregorio Juan Mc Callum, Ignacio Smith, Federico Javier Wolman, Alexandra Marisa Targovnik & María Victoria Miranda

  2. Instituto de Nanobiotecnología (NANOBIOTEC), CONICET-Universidad de Buenos Aires, Junín 956, 1113, Buenos Aires, Argentina

    Mariana Bernadett Arregui, Gregorio Juan Mc Callum, Ignacio Smith, Federico Javier Wolman, Alexandra Marisa Targovnik & María Victoria Miranda

  3. Unidad de Transferencia Genética, Instituto de Oncología “Ángel H. Roffo”, Universidad de Buenos Aires, Av. San Martín 5481, 1417, Buenos Aires, Argentina

    Marcela Solange Villaverde

  4. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina

    Marcela Solange Villaverde

Authors
  1. Mariana Bernadett Arregui
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  2. Gregorio Juan Mc Callum
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  3. Ignacio Smith
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  4. Marcela Solange Villaverde
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  5. Federico Javier Wolman
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  6. Alexandra Marisa Targovnik
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  7. María Victoria Miranda
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Contributions

MBA worked out almost all of the technical details and performed the numerical calculations; GJM and IS contributed with analytical methods; MSV performed antitumor activity assays; FJW contributed in downstream processing; AMT contributed in discussion; MVM was involved in planning and supervised the work. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to María Victoria Miranda.

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Arregui, M.B., Mc Callum, G.J., Smith, I. et al. Production of biologically active feline interferon beta in insect larvae using a recombinant baculovirus. 3 Biotech 8, 341 (2018). https://doi.org/10.1007/s13205-018-1369-x

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