Open AccessDESIGN OF HEPATITIS E VIRUS GENOTYPE 1 RECOMBINANT CAPSID PROTEIN: CLONING, EXPRESSION, PURIFICATION, EVALUATION OF THE ANTIGENIC PROPERTIES
Author(s) -
Г. И. Алаторцева,
Александр Викторович Сидоров,
Л. Н. Нестеренко,
L. N. Luhverchik,
В. В. Доценко,
И. И. Амиантова,
V. Yu. Kabargina,
А. В. Милованова,
Д С Воробьев,
Yulia Ammour,
В. М. Блинов,
А. З. Нурматов,
Zuridin Nurmatov,
D. A. Baiyzbekova,
Omor Tilegenovich Kasymov,
Karen K. Kyuregyan,
М. И. Михайлов,
С. В. Жаворонок,
В. В. Зверев
Publication year - 2017
Publication title -
žurnal mikrobiologii, èpidemiologii i immunobiologii
Language(s) - English
Resource type - Journals
eISSN - 2686-7613
pISSN - 0372-9311
DOI - 10.36233/0372-9311-2017-6-72-80
Subject(s) - recombinant dna , capsid , virology , hepatitis e virus , biology , microbiology and biotechnology , escherichia coli , genotype , serology , antigen , virus , antibody , gene , biochemistry , genetics , immunology
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.