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Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein

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Herpes Simplex Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2060))

Abstract

In the previous chapter, we describe the engineering of a HSV-BAC genome by galK recombineering. Here we describe the procedures to reconstitute, or regenerate, the replicating recombinant virus, and the methods to purify it and characterize it for the correct expression of the transgene. We present the example of R-115, a recombinant expressing murine interleukin 12 (mIL12) from the US1–US2 intergenic region. A specific method for the production of highly purified virions by iodixanol gradient, suitable for in vivo applications, is also detailed.

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References

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Acknowledgments

This work was supported by European Research Council (ERC) Advanced Grant number 340060, VII framework program to G.C.-F., by RFO (University of Bologna) to L.M. and T.G, and by Fondi Pallotti to T.G.

Competing interests: G.C.-F. owns shares in Nouscom Srl. B.P. is currently an employee of Nouscom Srl. G.C.-F. and L.M. receive equity payments from Amgen. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Department of Experimental, Diagnostic and Specialty Medicine, University of Bologna, Bologna, Italy

    Andrea Vannini, Valentina Gatta, Valerio Leoni, Simona Pepe, Gabriella Campadelli-Fiume & Tatiana Gianni

  2. Nouscom Srl, Rome, Italy

    Biljana Petrovic

  3. Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy

    Laura Menotti

Authors
  1. Andrea Vannini
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  2. Biljana Petrovic
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  3. Valentina Gatta
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  4. Valerio Leoni
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  5. Simona Pepe
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  6. Laura Menotti
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  7. Gabriella Campadelli-Fiume
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  8. Tatiana Gianni
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Corresponding author

Correspondence to Gabriella Campadelli-Fiume .

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Editors and Affiliations

  1. Department of Biomedical Sciences, Macquarie University, Sydney, NSW, Australia

    Russell J. Diefenbach

  2. Institute of Virology, University of Zurich, Zürich, Switzerland

    Cornel Fraefel

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Vannini, A. et al. (2020). Rescue, Purification, and Characterization of a Recombinant HSV Expressing a Transgenic Protein. In: Diefenbach, R., Fraefel, C. (eds) Herpes Simplex Virus . Methods in Molecular Biology, vol 2060. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9814-2_8

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  • DOI: https://doi.org/10.1007/978-1-4939-9814-2_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9813-5

  • Online ISBN: 978-1-4939-9814-2

  • eBook Packages: Springer Protocols

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