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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.
Mardanova, Eugenia S; Kotlyarov, Roman Y; Stuchinskaya, Maya D; Nikolaeva, Lyudmila I; Zahmanova, Gergana; Ravin, Nikolai V.
  • Mardanova ES; Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia.
  • Kotlyarov RY; Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, 119071 Moscow, Russia.
  • Stuchinskaya MD; N.F. Gamaleya National Research Centre of Epidemiology and Microbiology, Ministry of Health, 123098 Moscow, Russia.
  • Nikolaeva LI; N.F. Gamaleya National Research Centre of Epidemiology and Microbiology, Ministry of Health, 123098 Moscow, Russia.
  • Zahmanova G; Department of Plant Physiology and Molecular Biology, University of Plovdiv, 4000 Plovdiv, Bulgaria.
  • Ravin NV; Center of Plant Systems Biology and Biotechnology, 4000 Plovdiv, Bulgaria.
Int J Mol Sci ; 23(24)2022 Dec 10.
Article in English | MEDLINE | ID: covidwho-2155136
ABSTRACT
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' 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 300-400 µg 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 µg per gram of fresh leaf tissue. The fusion protein HEV/RBD, targeted to the endoplasmic reticulum, was expressed at about 80-100 µg per gram of fresh leaf tissue; the yield after purification was up to 20 µg 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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Hepatitis E virus / Capsid Proteins / Artificial Virus-Like Particles Topics: Vaccines Limits: Humans Language: English Year: 2022 Document Type: Article Affiliation country: Ijms232415684

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Hepatitis E virus / Capsid Proteins / Artificial Virus-Like Particles Topics: Vaccines Limits: Humans Language: English Year: 2022 Document Type: Article Affiliation country: Ijms232415684