{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,12]],"date-time":"2026-01-12T22:08:47Z","timestamp":1768255727548,"version":"3.49.0"},"reference-count":77,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2022,12,10]],"date-time":"2022-12-10T00:00:00Z","timestamp":1670630400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012190","name":"Ministry of Science and Higher Education of the Russian Federation","doi-asserted-by":"publisher","award":["075-15-2022-318"],"award-info":[{"award-number":["075-15-2022-318"]}],"id":[{"id":"10.13039\/501100012190","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Capsid protein of Hepatitis E virus (HEV) is capable of self-assembly into virus-like particles (VLPs) when expressed in Nicotiana benthamiana plants. Such VLPs could be used as carriers of antigens for vaccine development. In this study, we obtained VLPs based on truncated coat protein of HEV bearing the M2e peptide of Influenza A virus or receptor-binding domain of SARS-CoV-2 spike glycoprotein (RBD). We optimized the immunogenic epitopes\u2019 presentation by inserting them into the protruding domain of HEV ORF2 at position Tyr485. The fusion proteins were expressed in Nicotiana benthamiana plants using self-replicating potato virus X (PVX)-based vector. The fusion protein HEV\/M2, targeted to the cytosol, was expressed at the level of about 300\u2013400 \u03bcg per gram of fresh leaf tissue and appeared to be soluble. The fusion protein was purified using metal affinity chromatography under native conditions with the final yield about 200 \u03bcg per gram of fresh leaf tissue. The fusion protein HEV\/RBD, targeted to the endoplasmic reticulum, was expressed at about 80\u2013100 \u03bcg per gram of fresh leaf tissue; the yield after purification was up to 20 \u03bcg per gram of fresh leaf tissue. The recombinant proteins HEV\/M2 and HEV\/RBD formed nanosized virus-like particles that could be recognized by antibodies against inserted epitopes. The ELISA assay showed that antibodies of COVID-19 patients can bind plant-produced HEV\/RBD virus-like particles. This study shows that HEV capsid protein is a promising carrier for presentation of foreign antigen.<\/jats:p>","DOI":"10.3390\/ijms232415684","type":"journal-article","created":{"date-parts":[[2022,12,12]],"date-time":"2022-12-12T01:42:27Z","timestamp":1670809347000},"page":"15684","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["High-Yield Production of Chimeric Hepatitis E Virus-Like Particles Bearing the M2e Influenza Epitope and Receptor Binding Domain of SARS-CoV-2 in Plants Using Viral Vectors"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2148-7658","authenticated-orcid":false,"given":"Eugenia S.","family":"Mardanova","sequence":"first","affiliation":[{"name":"Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4387-2592","authenticated-orcid":false,"given":"Roman Y.","family":"Kotlyarov","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8544-7482","authenticated-orcid":false,"given":"Maya D.","family":"Stuchinskaya","sequence":"additional","affiliation":[{"name":"N.F. Gamaleya National Research Centre of Epidemiology and Microbiology, Ministry of Health, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1323-5568","authenticated-orcid":false,"given":"Lyudmila I.","family":"Nikolaeva","sequence":"additional","affiliation":[{"name":"N.F. Gamaleya National Research Centre of Epidemiology and Microbiology, Ministry of Health, 123098 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7667-9924","authenticated-orcid":false,"given":"Gergana","family":"Zahmanova","sequence":"additional","affiliation":[{"name":"Department of Plant Physiology and Molecular Biology, University of Plovdiv, 4000 Plovdiv, Bulgaria"},{"name":"Center of Plant Systems Biology and Biotechnology, 4000 Plovdiv, Bulgaria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1456-1832","authenticated-orcid":false,"given":"Nikolai V.","family":"Ravin","sequence":"additional","affiliation":[{"name":"Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Liu, H., and Timko, M.P. 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