Applied Microbiology and Biotechnology

, Volume 103, Issue 18, pp 7491–7504 | Cite as

Functional characterization of a plant-produced infectious bursal disease virus antigen fused to the constant region of avian IgY immunoglobulins

  • Emile Rage
  • Charifa Drissi Touzani
  • Carla Marusic
  • Chiara Lico
  • Thomas Göbel
  • Alessio Bortolami
  • Francesco Bonfante
  • Anna Maria Salzano
  • Andrea Scaloni
  • Siham Fellahi
  • Mohammed El Houadfi
  • Marcello DoniniEmail author
  • Selene Baschieri
Biotechnological products and process engineering


Infectious bursal disease virus (IBDV) is the cause of an economically important highly contagious disease of poultry, and vaccines are regarded as the most beneficial interventions for its prevention. In this study, plants were used to produce a recombinant chimeric IBDV antigen for the formulation of an innovative subunit vaccine. The fusion protein (PD-FcY) was designed to combine the immunodominant projection domain (PD) of the viral structural protein VP2 with the constant region of avian IgY (FcY), which was selected to enhance antigen uptake by avian immune cells. The gene construct encoding the fusion protein was transiently expressed in Nicotiana benthamiana plants and an extraction/purification protocol was set up, allowing to reduce the contamination by undesired plant compounds/proteins. Mass spectrometry analysis of the purified protein revealed that the glycosylation pattern of the FcY portion was similar to that observed in native IgY, while in vitro assays demonstrated the ability of PD-FcY to bind to the avian immunoglobulin receptor CHIR-AB1. Preliminary immunization studies proved that PD-FcY was able to induce the production of protective anti-IBDV-VP2 antibodies in chickens. In conclusion, the proposed fusion strategy holds promises for the development of innovative low-cost subunit vaccines for the prevention of avian viral diseases.


CHIR-AB1 Glycosylation IgY IBDV Molecular farming Veterinary vaccine 



The authors thank Prof. J. R. Caston for providing the rabbit anti-VP2 polyclonal serum.


This study was funded by AVIAMED project through the ERA-NET ARIMNet2 2015 Call by the following funding agencies: Italian Ministry of Agricultural, Food and Forestry Policies (MIPAAF), and Ministry of Higher Education, Scientific Research and Professional Training of Morocco (MESRSFC). ARIMNet2 (ERA-NET) has received funding from the European Union’s Seventh Framework Programme for research, technological development, and demonstration under grant agreement no. 618127/182.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Animal experiment procedures were conducted in strict accordance with the Decree of the Italian Ministry of Health n. 26 of 4 March, 2014, on the protection of animals used for scientific purposes, implementing Directive 2010/63/EU, and approved by IZSVe’s Ethics Committee or in accordance with European and French legislations on laboratory animal care and use (French Decree 2001-464 and European Directive CEE86/609) and animal protocols approved by the Ethics Committee “Sciences et santé animale,” committee number 115. The animals were kept within the animal facilities (biosafety level 2) of IZSVe or of Agronomy and Veterinary Institute Hassan II.

This article does not contain any studies with human participants performed by any of the authors.

Supplementary material

253_2019_9992_MOESM1_ESM.pdf (1.4 mb)
ESM 1 (PDF 1477 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Emile Rage
    • 1
  • Charifa Drissi Touzani
    • 2
  • Carla Marusic
    • 1
  • Chiara Lico
    • 1
  • Thomas Göbel
    • 3
  • Alessio Bortolami
    • 4
  • Francesco Bonfante
    • 4
  • Anna Maria Salzano
    • 5
  • Andrea Scaloni
    • 5
  • Siham Fellahi
    • 2
  • Mohammed El Houadfi
    • 2
  • Marcello Donini
    • 1
    Email author
  • Selene Baschieri
    • 1
  1. 1.Laboratory of BiotechnologyENEA Casaccia Research CenterRomeItaly
  2. 2.Unité de Pathologie Aviaire, Département de Pathologie et Santé Publique VétérinaireIAV Hassan IIRabatMorocco
  3. 3.Department of Veterinary SciencesLMU MunichMünchenGermany
  4. 4.Division of Comparative Biomedical ScienceIstituto Zooprofilattico Sperimentale delle VenezieLegnaroItaly
  5. 5.Proteomics & Mass Spectrometry Laboratory, ISPAAMNational Research CouncilNapoliItaly

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