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 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.

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Publication Details

Journal
Journal of microbiology epidemiology immunobiology
Published
2024-11-18
DOI
https://doi.org/10.36233/0372-9311-552
Citations
4
Primary Topic
Viral gastroenteritis research and epidemiology
Type
article
Field-Weighted Citation Impact
1.56
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article

Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties

Д. В. Новиков, В. А. Лапин, Е. В. Мохонова, Maria I. Tsyganova et al.
4 citations
Journal of microbiology epidemiology immunobiology
Viral gastroenteritis research and epidemiology
1.56
article

Production of recombinant norovirus VP1 protein and its antigenic and immunogenic properties

Д. В. Новиков, В. А. Лапин, Е. В. Мохонова, Maria I. Tsyganova, Novikov Vv, Dmitry A. Melentyev, Dmitry E. Zaitsev
article en
4 citations

Abstract

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.

Journal of microbiology epidemiology immunobiologyVol. 101(5)
Nizhniy Novgorod Research Institute of Epidemiology and Microbiology named after Academician I.N. Blokhina (RU)
Good health and well-being
Openalex Percentile: Top 16%
Viral gastroenteritis research and epidemiology
1.56
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