DESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT CAPSID PROTEIN: CLONING, EXPRESSION, PURIFICATION, EVALUATION OF THE ANTIGENIC PROPERTIES

G. I. Alatortseva; Алаторцева Г. И.; A. V. Sidorov; Сидоров А. В.; L. N. Nesterenko; Нестеренко Л. Н.; L. N. Luhverchik; Лухверчик Л. Н.; V. V. Dotsenko; Доценко В. B.; I. I. Amiantova; Амиантова И. И.; V. Yu. Kabargina; Кабаргина В. Ю.; A. V. Milovanova; Милованова А. В.; D. S. Vorobev; Воробьев Д. С.; Yu. I. Ammur; Аммур Ю. И.; V. M. Blinov; Блинов В. М.; A. Z. Nurmatov; Нурматов А. З.; Z. Sh. Nurmatov; Нурматов З. Ш.; D. A. Baiyzbekova; Байызбекова Д. А.; O. T. Kasymov; Касымов О. Т.; K. K. Kyuregyan; Кюрегян К. К.; M. I. Mikhailov; Михайлов М. И.; S. V. Zhavoronok; Жаворонок С. В.; V. V. Zverev; Зверев В. В.

doi:10.36233/0372-9311-2017-6-72-80

DESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT CAPSID PROTEIN: CLONING, EXPRESSION, PURIFICATION, EVALUATION OF THE ANTIGENIC PROPERTIES

Authors: Alatortseva G.I.¹, Sidorov A.V.¹, Nesterenko L.N.¹, Luhverchik L.N.¹, Dotsenko V.V.¹, Amiantova I.I.¹, Kabargina V.Y.¹, Milovanova A.V.¹, Vorobev D.S.¹, Ammur Y.I.¹, Blinov V.M.¹, Nurmatov A.Z.², Nurmatov Z.S.², Baiyzbekova D.A.², Kasymov O.T.², Kyuregyan K.K.³, Mikhailov M.I.³, Zhavoronok S.V.⁴, Zverev V.V.¹
Affiliations:
1. Mechnikov Research Institute of Vaccines and Sera
2. Scientific Production Association «Preventive Medicine»
3. Mechnikov Research Institute of Vaccines and Sera, Russian Medical Academy of Continuous Professional Education
4. Belarusian State Medical University
Issue: Vol 94, No 6 (2017)
Pages: 72-80
Section: Articles
URL: https://microbiol.crie.ru/jour/article/view/233
DOI: https://doi.org/10.36233/0372-9311-2017-6-72-80
ID: 233

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Abstract

Aim. The development of the hepatitis E virus (HEV) genotype 1 recombinant capsid protein. Materials and methods. Escherichia coli strains, plasmid vectors, serological and clinical samples, ELISA reagent kits, molecular biological, bioinformatic, biotechnological, biochemical and serological methods. Results. Using HEV genotype 1 DNA copy of subgenomic virus RNA we made E.coli strains producing recombinabt capsid protein, containing C-terminal fragment of ORF2 protein fused to E.coli beta-galactosidase. Recombinant protein ORF2 had been isolated from the inclusion bodies of the E.coli biomass and purified by size exclusion chromatography. By Western blotting it had been shown specific interaction of the recombinant polypeptide with anti-HEV IgG from pool of positive sera. Antigenic specificity of the recombinant polypeptide had been confirmed by enzyme-linked immunosorbent assay with sera of hepatitis E patients and reference groups: healthy donors, patients with hepatitis А, В, C, infectious mononucleosis and cytomegalovirus infection, HIV-infected patients. Conclusion. HEV genotype 1 ORF2 recombinant antigen had been developed, and its possible use in diagnostic tests had been experimentally shown.

Keywords

hepatitis E virus, genotype 1 HEV, orf2 gene, capsid protein, recombinant ORF2 antigen, ELISA, Western blotting

References

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DESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT CAPSID PROTEIN: CLONING, EXPRESSION, PURIFICATION, EVALUATION OF THE ANTIGENIC PROPERTIES

Full Text

Abstract

Keywords

About the authors

References

Supplementary files

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