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Generation of recombinant VP3 protein of infectious bursal disease virus in three different expression systems, antigenic analysis of the obtained polypeptides and development of an ELISA test

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Abstract

Infectious bursal disease virus (IBDV), which infects young chickens, is one of the most important pathogens that harm the poultry industry. Evaluation of the immune status of birds before and after vaccination is of great importance for controlling the disease caused by this virus. Therefore, the development of low-cost and easy-to-manufacture test systems for IBDV antibody detection remains an urgent issue. In this study, three expression systems (bacteria, yeast, and human cells) were used to produce recombinant VP3 protein of IBDV. VP3 is a group-specific antigen and hence may be a good candidate for use in diagnostic tests. Comparison of the antigenic properties of the obtained polypeptides showed that the titres of antibodies raised in chickens against bacteria- or human-cell-derived recombinant VP3 were high, whereas the antibody level against yeast-derived recombinant VP3 was low. The results of an enzyme-linked immunosorbent assay (ELISA) of sera from IBDV-infected chickens demonstrated that the recombinant VP3 produced in E. coli would be the best choice for use in test systems.

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Acknowledgements

We thank Daria Matyushkina (Federal Research and Clinical Centre of Physical-Chemical Medicine, Moscow, Russia) for the MALDI-TOF analysis. We also thank Tatyana A. Akopian and Oksana V. Selezneva (Federal Research and Clinical Centre of Physical-Chemical Medicine, Moscow, Russia) for the Sanger dideoxy sequencing of genetic constructs. We thank the Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency for the expertise and guidance in genetic engineering.

Funding

This work was supported by the Russian Science Foundation (project 16-16-04051).

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Authors and Affiliations

  1. Federal Research and Clinical Center of Physical-Chemical Medicine of Federal Medical Biological Agency, Moscow, Russian Federation

    Dmitriy A. Shirokov, Valentin A. Manuvera, Olga A. Miroshina & Vassili N. Lazarev

  2. K. I. Skryabin Moscow State Academy of Veterinary Medicine and Biotechnology, Moscow, Russian Federation

    Dmitriy A. Shirokov

  3. Moscow Institute of Physics and Technology, Dolgoprudny, Russian Federation

    Valentin A. Manuvera, Olga A. Miroshina & Vassili N. Lazarev

  4. All-Russian Research Veterinary Institute of Poultry Science, Branch of All-Russian Research and Technological Poultry Institute of the Russian Academy of Sciences, Sergiyev Posad, Russian Federation, St. Petersburg, Russian Federation

    Valentin A. Manuvera, Alexandr S. Dubovoi, Galina N. Samuseva & Margarita E. Dmitrieva

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  1. Dmitriy A. Shirokov
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  2. Valentin A. Manuvera
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Correspondence to Dmitriy A. Shirokov.

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Ethical statement

Care and use of the laboratory animals involved in this research was carried out in accordance with EU Directive 2010/63/EU for animal experiments and FELASA recommendations (e.g. Guidelines for the Veterinary Care of Laboratory Animals [2008]). The animal use protocol was reviewed and approved by the Ethics Committee of the All-Russian Research Veterinary Institute of Poultry Science.

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Shirokov, D.A., Manuvera, V.A., Miroshina, O.A. et al. Generation of recombinant VP3 protein of infectious bursal disease virus in three different expression systems, antigenic analysis of the obtained polypeptides and development of an ELISA test. Arch Virol 165, 1611–1620 (2020). https://doi.org/10.1007/s00705-020-04650-2

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