Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties
Introduction. The importance of noroviruses in human infectious pathology and the danger of large epidemic outbreaks in organized groups determine the need to develop means of specific prevention of infection. The aim of the study was to obtain recombinant norovirus VP1 protein and analyze its im...
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| Format: | Article |
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Central Research Institute for Epidemiology
2024-11-01
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| Series: | Журнал микробиологии, эпидемиологии и иммунобиологии |
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| Online Access: | https://microbiol.crie.ru/jour/article/viewFile/18687/1522 |
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| author | Vladislav A. Lapin Dmitry V. Novikov Ekaterina V. Mokhonova Dmitry A. Melentyev Maria I. Tsyganova Dmitry E. Zaitsev Viktor V. Novikov |
| author_facet | Vladislav A. Lapin Dmitry V. Novikov Ekaterina V. Mokhonova Dmitry A. Melentyev Maria I. Tsyganova Dmitry E. Zaitsev Viktor V. Novikov |
| author_sort | Vladislav A. Lapin |
| collection | DOAJ |
| description | Introduction. The importance of noroviruses in human infectious pathology and the danger of large epidemic outbreaks in organized groups determine the need to develop means of specific prevention of infection.
The aim of the study was to obtain recombinant norovirus VP1 protein and analyze its immunogenic and antigenic properties .
Materials and methods. Computer analysis of nucleotide and amino acid sequences, molecular cloning, polymerase chain reaction, electrophoresis of nucleic acids in agarose gel and proteins in polacrylamide gel, affinity chromatography, enzyme immunoassay.
Results and discussion. A genetic construct encoding recombinant VP1 of the GII genotype norovirus with codons optimized for highly effective expression in Escherichia coli has been created. The strain of E. coli Rosetta 2 (DE3) has been transformed by genetic construct. VP1 expression was carried out in E. coli cells, conditions for its production, purification and renaturation were optimized. A purified soluble recombinant VP1 protein forms virus-like particles with a diameter of 30–50 nm. Immunization of BALB/c mice by protein lead to antibodies production with a titer greater than 1 : 1000. When evaluating antigenic properties, it was shown that human IgG, IgM, and IgA antibodies interact with recombinant VP1. The total antibody detection rate was 47,4%. The results indicate the possibility of using recombinant VP1 for development of domestic vaccine for the prevention of norovirus infection. |
| format | Article |
| id | doaj-art-711ef7d6602b43fdbb7870ded3d83c48 |
| institution | Kabale University |
| issn | 0372-9311 2686-7613 |
| language | Russian |
| publishDate | 2024-11-01 |
| publisher | Central Research Institute for Epidemiology |
| record_format | Article |
| series | Журнал микробиологии, эпидемиологии и иммунобиологии |
| spelling | doaj-art-711ef7d6602b43fdbb7870ded3d83c482025-01-09T20:09:08ZrusCentral Research Institute for EpidemiologyЖурнал микробиологии, эпидемиологии и иммунобиологии0372-93112686-76132024-11-01101566166710.36233/0372-9311-5522787Production of recombinant norovirus VP1 protein and its antigenic and immunogenic propertiesVladislav A. Lapin0https://orcid.org/0000-0002-5905-5722Dmitry V. Novikov1https://orcid.org/0000-0001-7049-6935Ekaterina V. Mokhonova2https://orcid.org/0000-0002-9742-7646Dmitry A. Melentyev3https://orcid.org/0000-0002-2441-6874Maria I. Tsyganova4https://orcid.org/0000-0002-2811-6844Dmitry E. Zaitsev5https://orcid.org/0000-0002-7663-6924Viktor V. Novikov6https://orcid.org/0000-0002-2449-7213Academician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyAcademician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyAcademician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyAcademician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyAcademician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyAcademician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyAcademician I.N. Blokhina Nizhny Novgorod Scientific Research Institute of Epidemiology and MicrobiologyIntroduction. The importance of noroviruses in human infectious pathology and the danger of large epidemic outbreaks in organized groups determine the need to develop means of specific prevention of infection. The aim of the study was to obtain recombinant norovirus VP1 protein and analyze its immunogenic and antigenic properties . Materials and methods. Computer analysis of nucleotide and amino acid sequences, molecular cloning, polymerase chain reaction, electrophoresis of nucleic acids in agarose gel and proteins in polacrylamide gel, affinity chromatography, enzyme immunoassay. Results and discussion. A genetic construct encoding recombinant VP1 of the GII genotype norovirus with codons optimized for highly effective expression in Escherichia coli has been created. The strain of E. coli Rosetta 2 (DE3) has been transformed by genetic construct. VP1 expression was carried out in E. coli cells, conditions for its production, purification and renaturation were optimized. A purified soluble recombinant VP1 protein forms virus-like particles with a diameter of 30–50 nm. Immunization of BALB/c mice by protein lead to antibodies production with a titer greater than 1 : 1000. When evaluating antigenic properties, it was shown that human IgG, IgM, and IgA antibodies interact with recombinant VP1. The total antibody detection rate was 47,4%. The results indicate the possibility of using recombinant VP1 for development of domestic vaccine for the prevention of norovirus infection.https://microbiol.crie.ru/jour/article/viewFile/18687/1522norovirusvp1virus-like particlesvaccine |
| spellingShingle | Vladislav A. Lapin Dmitry V. Novikov Ekaterina V. Mokhonova Dmitry A. Melentyev Maria I. Tsyganova Dmitry E. Zaitsev Viktor V. Novikov Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties Журнал микробиологии, эпидемиологии и иммунобиологии norovirus vp1 virus-like particles vaccine |
| title | Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties |
| title_full | Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties |
| title_fullStr | Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties |
| title_full_unstemmed | Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties |
| title_short | Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties |
| title_sort | production of recombinant norovirus vp1 protein and its antigenic and immunogenic properties |
| topic | norovirus vp1 virus-like particles vaccine |
| url | https://microbiol.crie.ru/jour/article/viewFile/18687/1522 |
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