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High-Throughput Production of Influenza Virus-Like Particle (VLP) Array by Using VLP-factory™, a MultiBac Baculoviral Genome Customized for Enveloped VLP Expression

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High-Throughput Protein Production and Purification

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2025))

Abstract

Baculovirus-based expression of proteins in insect cell cultures has emerged as a powerful technology to produce complex protein biologics for many applications ranging from multiprotein complex structural biology to manufacturing of therapeutic proteins including virus-like particles (VLPs). VLPs are protein assemblies that mimic live viruses but typically do not contain any genetic material, and therefore are safe and attractive alternatives to life attenuated or inactivated viruses for vaccination purposes. MultiBac is an advanced baculovirus expression vector system (BEVS) which consists of an engineered viral genome that can be customized for tailored applications. Here we describe the creation of a MultiBac-based VLP-factory™, based on the M1 capsid protein from influenza, and its application to produce in a parallelized fashion an array of influenza-derived VLPs containing functional mutations in influenza hemagglutinin (HA) thought to modulate the immune response elicited by the VLP.

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Acknowledgements

We thank all members of the Berger and Schaffitzel laboratories for helpful discussions, as well as Rob Ruigrok, Thibaut Crepin, and Darren Hart for expert insight in influenza biology. We are grateful to Karin Huard for assistance with gradient preparation, and Yan Nie for introduction to negative-stain EM. This work was supported by the European Commission Framework Programme 7 projects ComplexINC (contract nr. 279039) and SynSignal (contract nr. 613879).

Competing Financial Interest Statement

The authors declare competing financial interest. Parts of the technology here described are subject of international patent EP2403940 and licensed exclusively to Geneva Biotech SARL.

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Authors and Affiliations

  1. The European Molecular Biology Laboratory (EMBL), Grenoble Cedex 9, France

    Duygu Sari-Ak & Petra Drncova

  2. AimVion AS, Aarhus, Denmark

    Shervin Bahrami

  3. Department of Biomedicine, Bartholins Allé 6, University of Aarhus, Aarhus C, Denmark

    Magdalena J. Laska

  4. Geneva Biotech SARL, Geneva, Switzerland

    Daniel J. Fitzgerald

  5. School of Biochemistry and Bristol Synthetic Biology Centre BrisSynBio, University Walk, University of Bristol, Clifton, UK

    Christiane Schaffitzel & Imre Berger

  6. Imophoron Ltd, Unit DX, St Philips Central, Bristol, UK

    Frederic Garzoni

Authors
  1. Duygu Sari-Ak
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  2. Shervin Bahrami
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  3. Magdalena J. Laska
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  4. Petra Drncova
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  5. Daniel J. Fitzgerald
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  6. Christiane Schaffitzel
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  7. Frederic Garzoni
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  8. Imre Berger
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Corresponding authors

Correspondence to Frederic Garzoni or Imre Berger .

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Editors and Affiliations

  1. Architecture et Fonction des Macromolécules Biologiques (AFMB), Unité Mixte de Recherche (UMR) 7257, Centre National de la Recherche Scientifique (CNRS), Aix-Marseille Université, Marseille cedex 9, France

    Renaud Vincentelli

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Sari-Ak, D. et al. (2019). High-Throughput Production of Influenza Virus-Like Particle (VLP) Array by Using VLP-factory™, a MultiBac Baculoviral Genome Customized for Enveloped VLP Expression. In: Vincentelli, R. (eds) High-Throughput Protein Production and Purification. Methods in Molecular Biology, vol 2025. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9624-7_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9624-7_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9623-0

  • Online ISBN: 978-1-4939-9624-7

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