Construction of norovirus (Caliciviridae: Norovirus) virus-like particles containing VP1 of the Echovirus 30 (Picornaviridae: Enterovirus: Enterovirus B)
- Authors: Novikov D.V.1, Melentev D.A.1,2, Mokhonov V.V.1, Kashnikov A.Y.1, Novikova N.A.1, Lapin V.A.1,2, Mokhonova E.V.1, Novikov V.V.1,2
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Affiliations:
- FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
- FSAEI HE «National Research Lobachevsky State University of Nizhny Novgorod» of the Ministry of Education and Science of Russia
- Issue: Vol 66, No 5 (2021)
- Pages: 383-389
- Section: ORIGINAL RESEARCH
- URL: https://journals.rcsi.science/0507-4088/article/view/118183
- DOI: https://doi.org/10.36233/0507-4088-79
- ID: 118183
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Abstract
Introduction. Enterovirus (nonpolio) infection is widespread all over the world, registered as sporadic cases and large-scale outbreaks and can cause severe lesions such as serous meningitis. Epidemiological studies have shown that enterovirus (Picornaviridae; Enterovirus) variant Echovirus 30 (E30) is the most frequently detected variant in patients with enterovirus meningitis in the Russian Federation. However, no vaccines to prevent the disease caused by this pathogen have been developed so far. One of the promising modern trends in terms of creating vaccine preparations is the use of virus-like particles (VLPs), including chimeric ones containing the biological structures of viruses belonging to different species.
The aim of this work was to obtain norovirus (Caliciviridae; Norovirus) VLPs displaying enterovirus Echovirus E30 full-length VP1 on the surface.
Material and methods. The nucleotide sequences of VP1 protein of norovirus genotype GII.4 and VP1 E30 of genotype h circulating in Russia were used. The SN-VP1E30 protein was constructed, in which the shell (S) and the hinge regions of the norovirus VP1 are fused into one molecule with the full-length VP1 of the E30 virus. The protein was expressed in E. coli, purified using affinity chromatography, and characterized by polyacrylamide gel electrophoresis (PAGE) and immunoblotting. VLPs were visualized by electron microscopy.
Results. The S N-VP1E30 protein expressed in E. coli as insoluble form, so the conditions for SN-VP1E30 solublisation were defined. Sucrose has been shown to significantly increase the efficiency of renaturation. Electrophoretic mobility comparison of denatured and non-denatured SN-VP1E30 demonstrated that most monomers form high molecular weight compounds. Electron microscopy showed that renatured SN-VP1E30 spontaneously forms empty virus-like particles about 50 nm in diameter.
Conclusion. Chimeric protein SN-VP1E30 self-assemble into VLPs displaying the VP1 protein of E30 variant that is highly prevalent in Russia. Further immunological research is necessary to characterize VLPs potential for development of the vaccine for enteroviral meningitis prevention.
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##article.viewOnOriginalSite##About the authors
D. V. Novikov
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
Author for correspondence.
Email: novikov.dv75@mail.ru
ORCID iD: 0000-0001-7049-6935
Dmitry V. Novikov, Ph.D. (Biol.), Associate Professor, Lead Researcher of the Immunochemistry Laboratory
603950, Nizhny Novgorod, Russia
Russian FederationD. A. Melentev
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); FSAEI HE «National Research Lobachevsky State University of Nizhny Novgorod» of the Ministry of Education and Science of Russia
Email: fake@neicon.ru
ORCID iD: 0000-0002-2441-6874
603950, Nizhny Novgorod, Russia
Russian FederationV. V. Mokhonov
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
Email: fake@neicon.ru
ORCID iD: 0000-0002-8542-5723
603950, Nizhny Novgorod, Russia
Russian FederationA. Yu. Kashnikov
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
Email: fake@neicon.ru
ORCID iD: 0000-0003-1033-7347
603950, Nizhny Novgorod, Russia
Russian FederationN. A. Novikova
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
Email: fake@neicon.ru
ORCID iD: 0000-0002-3710-6648
603950, Nizhny Novgorod, Russia
Russian FederationV. A. Lapin
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); FSAEI HE «National Research Lobachevsky State University of Nizhny Novgorod» of the Ministry of Education and Science of Russia
Email: fake@neicon.ru
ORCID iD: 0000-0002-5905-5722
603950, Nizhny Novgorod, Russia
Russian FederationE. V. Mokhonova
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor)
Email: fake@neicon.ru
ORCID iD: 0000-0002-9742-7646
603950, Nizhny Novgorod, Russia
Russian FederationV. V. Novikov
FSBI «Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and Microbiology» of the Federal Service for Supervision of Consumer Rights Protection and Human Welfare (Rospotrebnadzor); FSAEI HE «National Research Lobachevsky State University of Nizhny Novgorod» of the Ministry of Education and Science of Russia
Email: fake@neicon.ru
ORCID iD: 0000-0002-2449-7213
603950, Nizhny Novgorod, Russia
Russian FederationReferences
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