DEVELOPMENT OF HEPATITIS E 3 GENOTYPE RECOMBINANT PROTEIN CAPSID OF: CLONING, EXPRESSION, PURIFICATION, EVALUATION OF THE ANTIGENIC PROPERTIES

• Published in: Žurnal mikrobiologii, ėpidemiologii i immunobiologii Vol. 96; no. 1; pp. 10 - 17
• Main Authors: Alatortseva, G. I., Sidorov, A. V., Nesterenko, L. N., Luhverchik, L. N., Dotsenko, V. V., Amiantova, I. I., Zhukina, M. V., Kabargina, V. Yu, Milovanova, A. V., Vorobev, D. S., Ammur, Yu. I., Mikhailov, M. I., Kyuregyan, K. K., Malinnikova, E. Yu, Zhavoronok, S. V., Blinov, V. M., Zverev, V. V.
• Format: Journal Article
• Language: English; Russian
• Published: Central Research Institute for Epidemiology 01.08.2019
• Subjects: capsid protein; elisa; hev genotype 3; orf2 gene; recombinant orf3 antigen; western blot
• ISSN: 0372-9311; 2686-7613
• DOI: 10.36233/0372-9311-2019-1-10-17

Aim. The development of the hepatitis E virus (HEV) genotype 3 recombinant capsid protein. Materials and methods. E.coli strains, plasmid vectors, serological and clinical samples, ELISA reagent kits, molecular biological, bioinformatic, biotechnological, biochemical and serological methods. Results. Using viruscontaining material from pigs of Belgorod region (Russian Federation) we made E.coli strains producing recombinant capsid protein, containing C-terminal of viral ORF2 protein fragment fused to E.coli β-galactosidase. Recombinant protein ORF2 had been isolated from the bacterial inclusion bodies and purified by size exclusion chromatography. Antigenic specificity of the recombinant polypeptide was confirmed by ELISA and Western blotting with sera of hepatitis E patients and reference groups (healthy donors, patients with hepatitis A, B, C, infectious mononucleosis, cytomegalovirus infection and HIV-infected patients). Conclusion. HEV genotype 3 ORF2 recombinant antigen had been developed, and the possibility to use it in diagnostic tests had been experimentally shown.