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
Abstract
1. Introduction
2. Results
2.1. Viral Vectors for the Expression of HEV Capsid Proteins Comprising M2e and RBD Peptides at the Tyr485 Position
2.2. Expression and Purification of HEV/M2e and HEV/RBD Proteins
2.3. Recombinant Proteins HEV/M2e and HEV/RBD form Virus-Like Particles
2.4. VLPs Formed by Recombinant Protein HEV/RBD Specifically Interacts with Sera of COVID-19 Patients
3. Discussion
4. Materials and Methods
4.1. Expression Vectors
4.2. Gene Cloning and Construction of Plasmids for Expression of HEV/M2e
4.3. Gene Cloning and Construction of Plasmids for Expression of HEV/RBD
4.4. Agroinfiltration of Nicotiana benthamiana Plants
4.5. SDS-PAGE and Western Blot Analyses
4.6. Isolation and Purification of Plant-Produced HEV/M2 and HEV/RBD Proteins
4.7. Nanoparticle Analysis
4.8. Serum Samples and ELISA
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
AMV | 5′-nontranslated region of RNA 4 of the alfalfa mosaic virus |
ER | endoplasmic reticulum |
GFP | Green fluorescent protein |
HEV | Hepatitis E virus |
M2e | extracellular domain of membrane protein M2 of influenza A virus |
PBS | Phosphate buffered saline |
PVX | Potato virus X |
RBD | Receptor Binding Domain of the S protein of SARS-CoV-2 |
SARS-CoV-2 | Severe acute respiratory syndrome coronavirus 2 |
SP | signal peptide |
VLP | virus-like particle |
References
- Liu, H.; Timko, M.P. Improving Protein Quantity and Quality-The Next Level of Plant Molecular Farming. Int. J. Mol. Sci. 2022, 23, 1326. [Google Scholar] [CrossRef]
- Zahmanova, G.; Takova, K.; Valkova, R.; Toneva, V.; Minkov, I.; Andonov, A.; Lukov, G.L. Plant-Derived Recombinant Vaccines against Zoonotic Viruses. Life 2022, 12, 156. [Google Scholar] [CrossRef]
- Shanmugaraj, B.; Bulaon, C.J.I.; Phoolcharoen, W. Plant Molecular Farming: A Viable Platform for Recombinant Biopharmaceutical Production. Plants 2020, 9, 842. [Google Scholar] [CrossRef]
- Sainsbury, F. Innovation in plant-based transient protein expression for infectious disease prevention and preparedness. Curr. Opin. Biotechnol. 2020, 61, 110–115. [Google Scholar] [CrossRef]
- Thuenemann, E.C.; Byrne, M.J.; Peyret, H.; Saunders, K.; Castells-Graells, R.; Ferriol, I.; Santoni, M.; Steele, J.F.C.; Ranson, N.A.; Avesani, L.; et al. A Replicating Viral Vector Greatly Enhances Accumulation of Helical Virus-Like Particles in Plants. Viruses 2021, 13, 885. [Google Scholar] [CrossRef]
- Nosaki, S.; Miura, K. Transient expression of recombinant proteins in plants. Methods Enzymol. 2021, 660, 193–203. [Google Scholar] [CrossRef]
- Lomonossoff, G.P.; DAoust, M.-A. Plant-Produced Biopharmaceuticals: A Case of Technical Developments Driving Clinical Deployment. Science 2016, 353, 1237–1240. [Google Scholar] [CrossRef]
- Kushnir, N.; Streatfield, S.J.; Yusibov, V. Virus-like Particles as a Highly Efficient Vaccine Platform: Diversity of Targets and Production Systems and Advances in Clinical Development. Vaccine 2012, 31, 58–83. [Google Scholar] [CrossRef]
- Rybicki, E.P. Plant-Based Vaccines against Viruses. Virol. J. 2014, 11, 205. [Google Scholar] [CrossRef]
- Márquez-Escobar, V.A.; Rosales-Mendoza, S.; Beltrán-López, J.I.; González-Ortega, O. Plant-Based Vaccines against Respiratory Diseases: Current Status and Future Prospects. Expert Rev. Vaccines 2017, 16, 137–149. [Google Scholar] [CrossRef]
- Chung, Y.H.; Church, D.; Koellhoffer, E.C.; Osota, E.; Shukla, S.; Rybicki, E.P.; Pokorski, J.K.; Steinmetz, N.F. Integrating plant molecular farming and materials research for next-generation vaccines. Nat. Rev. Mater. 2022, 7, 372–388. [Google Scholar] [CrossRef] [PubMed]
- Ward, B.J.; Makarkov, A.; Séguin, A.; Pillet, S.; Trépanier, S.; Dhaliwall, J.; Libman, M.D.; Vesikari, T.; Landry, N. Efficacy, Immunogenicity, and Safety of a Plant-Derived, Quadrivalent, Virus-like Particle Influenza Vaccine in Adults (18–64 Years) and Older Adults (≥65 Years): Two Multicentre, Randomised Phase 3 Trials. Lancet 2020, 396, 1491–1503. [Google Scholar] [CrossRef] [PubMed]
- Ward, B.J.; Gobeil, P.; Séguin, A.; Atkins, J.; Boulay, I.; Charbonneau, P.-Y.; Couture, M.; D’Aoust, M.-A.; Dhaliwall, J.; Finkle, C.; et al. Phase 1 Randomized Trial of a Plant-Derived Virus-like Particle Vaccine for COVID-19. Nat. Med. 2021, 27, 1071–10781. [Google Scholar] [CrossRef] [PubMed]
- Medicago. Available online: https://medicago.com/ (accessed on 4 November 2022).
- Webster, R.G.; Bean, W.J.; Gorman, O.T.; Chambers, T.M.; Kawaoka, Y. Evolution and Ecology of Influenza a Viruses. Microbiol. Rev. 1992, 56, 152–179. [Google Scholar] [CrossRef]
- Fiers, W.; De Filette, M.; Bakkouri, K.E.; Schepens, B.; Roose, K.; Schotsaert, M.; Birkett, A.; Saelens, X. M2e-Based Universal Influenza a Vaccine. Vaccine 2009, 27, 6280–6283. [Google Scholar] [CrossRef]
- Fiers, W.; De Filette, M.; Birkett, A.; Neirynck, S.; Min Jou, W. A “Universal” Human Influenza a Vaccine. Virus Res. 2004, 103, 173–176. [Google Scholar] [CrossRef]
- Ito, T.; Gorman, O.T.; Kawaoka, Y.; Bean, W.J.; Webster, R.G. Evolutionary Analysis of the Influenza a Virus M Gene with Comparison of the M1 and M2 Proteins. J. Virol. 1991, 65, 5491–5498. [Google Scholar] [CrossRef]
- Neirynck, S.; Deroo, T.; Saelens, X.; Vanlandschoot, P.; Jou, W.M.; Fiers, W. A Universal Influenza a Vaccine Based on the Extracellular Domain of the M2 Protein. Nat. Med. 1999, 5, 1157–1163. [Google Scholar] [CrossRef]
- Feng, J.; Zhang, M.; Mozdzanowska, K.; Zharikova, D.; Hoff, H.; Wunner, W.; Couch, R.B.; Gerhard, W. Influenza a Virus Infection Engenders a Poor Antibody Response against the Ectodomain of Matrix Protein 2. Virol. J. 2006, 3, 102. [Google Scholar] [CrossRef]
- De Filette, M.; Fiers, W.; Martens, W.; Birkett, A.; Ramne, A.; Löwenadler, B.; Lycke, N.; Jou, W.M.; Saelens, X. Improved Design and Intranasal Delivery of an M2e-Based Human Influenza a Vaccine. Vaccine 2006, 24, 6597–6601. [Google Scholar] [CrossRef]
- Mardanova, E.S.; Ravin, N.V. Plant-Produced Recombinant Influenza a Vaccines Based on the M2e Peptide. Curr. Pharm. Des. 2018, 24, 1317–1324. [Google Scholar] [CrossRef]
- Mardanova, E.S.; Kotlyarov, R.Y.; Kuprianov, V.V.; Stepanova, L.A.; Tsybalova, L.M.; Lomonosoff, G.P.; Ravin, N.V. Rapid high-yield expression of a candidate influenza vaccine based on the ectodomain of M2 protein linked to flagellin in plants using viral vectors. BMC Biotechnol. 2015, 15, 42. [Google Scholar] [CrossRef]
- Blokhina, E.A.; Mardanova, E.S.; Stepanova, L.A.; Tsybalova, L.M.; Ravin, N.V. Plant-Produced Recombinant Influenza a Virus Candidate Vaccine Based on Flagellin Linked to Conservative Fragments of M2 Protein and Hemagglutintin. Plants 2020, 9, 162. [Google Scholar] [CrossRef] [PubMed]
- Nemchinov, L.G.; Natilla, A. Transient Expression of the Ectodomain of Matrix Protein 2 (M2e) of Avian Influenza a Virus in Plants. Protein Expr. Purif. 2007, 56, 153–159. [Google Scholar] [CrossRef] [PubMed]
- Rasochova, L.; Radam, J.M.; Phelps, J.P.; Dang, N. Recombinant Flu Vaccines. Patent WO2007011904A2, 25 January 2007. [Google Scholar]
- Tyulkina, L.G.; Skurat, E.V.; Frolova, O.Y.; Komarova, T.V.; Karger, E.M.; Atabekov, I.G. New Viral Vector for Superproduction of Epitopes of Vaccine Proteins in Plants. Acta Nat. 2011, 3, 73–82. [Google Scholar] [CrossRef]
- Petukhova, N.; Gasanova, T.; Ivanov, P.; Atabekov, J. High-Level Systemic Expression of Conserved Influenza Epitope in Plants on the Surface of Rod-Shaped Chimeric Particles. Viruses 2014, 6, 1789–1800. [Google Scholar] [CrossRef] [PubMed]
- Petukhova, N.; Gasanova, T.; Stepanova, L.; Rusova, O.; Potapchuk, M.; Korotkov, A.; Skurat, E.; Tsybalova, L.; Kiselev, O.; Ivanov, P.; et al. Immunogenicity and Protective Efficacy of Candidate Universal Influenza A Nanovaccines Produced in Plants by Tobacco Mosaic Virus-Based Vectors. Curr. Pharm. Des. 2013, 19, 5587–5600. [Google Scholar] [CrossRef] [PubMed]
- Matić, S.; Rinaldi, R.; Masenga, V.; Noris, E. Efficient Production of Chimeric Human Papillomavirus 16 L1 Protein Bearing the M2e Influenza Epitope in Nicotiana Benthamiana Plants. BMC Biotechnol. 2011, 11, 106. [Google Scholar] [CrossRef]
- Ravin, N.V.; Kotlyarov, R.Y.; Mardanova, E.S.; Kuprianov, V.V.; Migunov, A.I.; Stepanova, L.A.; Tsybalova, L.M.; Kiselev, O.I.; Skryabin, K.G. Plant-Produced Recombinant Influenza Vaccine Based on Virus-like HBc Particles Carrying an Extracellular Domain of M2 Protein. Biochemistry 2012, 77, 33–40. [Google Scholar] [CrossRef]
- Thuenemann, E.C.; Lenzi, P.; Andrew, J.L.; Taliansky, M.; Bécares, M.; Zuñiga, S.; Enjuanes, L.; Zahmanova, G.G.; Minkov, I.N.; Matić, S.; et al. The use of transient expression systems for the rapid production of virus-like particles in plants. Curr. Pharm. Des. 2013, 19, 5564–5573. [Google Scholar] [CrossRef]
- Mardanova, E.S.; Blokhina, E.A.; Tsybalova, L.M.; Peyret, H.; Lomonossoff, G.P.; Ravin, N.V. Efficient Transient Expression of Recombinant Proteins in Plants by the Novel pEff Vector Based on the Genome of Potato Virus X. Front. Plant Sci. 2017, 8, 247. [Google Scholar] [CrossRef] [PubMed]
- Mbewana, S.; Mortimer, E.; Pêra, F.F.P.G.; Hitzeroth, I.I.; Rybicki, E.P. Production of H5N1 Influenza Virus Matrix Protein 2 Ectodomain Protein Bodies in Tobacco Plants and in Insect Cells as a Candidate Universal Influenza Vaccine. Front. Bioeng. Biotechnol. 2015, 3, 197. [Google Scholar] [CrossRef] [PubMed]
- Firsov, A.; Tarasenko, I.; Mitiouchkina, T.; Ismailova, N.; Shaloiko, L.; Vainstein, A.; Dolgov, S. High-Yield Expression of M2e Peptide of Avian Influenza Virus H5N1 in Transgenic Duckweed Plants. Mol. Biotechnol. 2015, 57, 653–661. [Google Scholar] [CrossRef] [PubMed]
- Firsov, A.; Tarasenko, I.; Mitiouchkina, T.; Shaloiko, L.; Kozlov, O.; Vinokurov, L.; Rasskazova, E.; Murashev, A.; Vainstein, A.; Dolgov, S. Expression and Immunogenicity of M2e Peptide of Avian Influenza Virus H5N1 Fused to Ricin Toxin B Chain Produced in Duckweed Plants. Front. Chem. 2018, 6, 22. [Google Scholar] [CrossRef]
- Walls, A.C.; Park, Y.-J.; Tortorici, M.A.; Wall, A.; McGuire, A.T.; Veesler, D. Structure, Function, and Antigenicity of the Sars-Cov-2 Spike Glycoprotein. Cell 2020, 181, 281–292. [Google Scholar] [CrossRef]
- Wrapp, D.; Wang, N.; Corbett, K.S.; Goldsmith, J.A.; Hsieh, C.-L.; Abiona, O.; Graham, B.S.; McLellan, J.S. Cryo-EM Structure of the 2019-NCoV Spike in the Prefusion Conformation. Science 2020, 367, 1260–1263. [Google Scholar] [CrossRef] [PubMed]
- Lan, J.; Ge, J.; Yu, J.; Shan, S.; Zhou, H.; Fan, S.; Zhang, Q.; Shi, X.; Wang, Q.; Zhang, L.; et al. Structure of the SARS-CoV-2 Spike Receptor-Binding Domain Bound to the ACE2 Receptor. Nature 2020, 581, 215–220. [Google Scholar] [CrossRef]
- Dai, L.; Gao, G.F. Viral Targets for Vaccines against COVID-19. Nat Rev Immunol. 2020, 21, 73–82. [Google Scholar] [CrossRef]
- Rattanapisit, K.; Shanmugaraj, B.; Manopwisedjaroen, S.; Purwono, P.B.; Siriwattananon, K.; Khorattanakulchai, N.; Hanittinan, O.; Boonyayothin, W.; Thitithanyanont, A.; Smith, D.R.; et al. Rapid Production of SARS-CoV-2 Receptor Binding Domain (RBD) and Spike Specific Monoclonal Antibody CR3022 in Nicotiana Benthamiana. Sci. Rep. 2020, 10, 17698. [Google Scholar] [CrossRef]
- Maharjan, P.M.; Cheon, J.; Jung, J.; Kim, H.; Lee, J.; Song, M.; Jeong, G.U.; Kwon, Y.; Shim, B.; Choe, S. Plant-Expressed Receptor Binding Domain of the SARS-CoV-2 Spike Protein Elicits Humoral Immunity in Mice. Vaccines 2021, 9, 978. [Google Scholar] [CrossRef]
- Mamedov, T.; Yuksel, D.; Ilgın, M.; Gurbuzaslan, I.; Gulec, B.; Yetiskin, H.; Uygut, M.A.; Islam Pavel, S.T.; Ozdarendeli, A.; Mammadova, G.; et al. Plant-Produced Glycosylated and in Vivo Deglycosylated Receptor Binding Domain Proteins of SARS-CoV-2 Induce Potent Neutralizing Responses in Mice. Viruses 2021, 13, 1595. [Google Scholar] [CrossRef] [PubMed]
- Siriwattananon, K.; Manopwisedjaroen, S.; Shanmugaraj, B.; Rattanapisit, K.; Phumiamorn, S.; Sapsutthipas, S.; Trisiriwanich, S.; Prompetchara, E.; Ketloy, C.; Buranapraditkun, S.; et al. Plant-Produced Receptor-Binding Domain of SARS-CoV-2 Elicits Potent Neutralizing Responses in Mice and Non-Human Primates. Front. Plant Sci. 2021, 12, 682953. [Google Scholar] [CrossRef] [PubMed]
- Mardanova, E.S.; Kotlyarov, R.Y.; Ravin, N.V. High-Yield Production of Receptor Binding Domain of SARS-CoV-2 Linked to Bacterial Flagellin in Plants Using Self-Replicating Viral Vector PEff. Plants 2021, 10, 2682. [Google Scholar] [CrossRef] [PubMed]
- Royal, J.M.; Simpson, C.A.; McCormick, A.A.; Phillips, A.; Hume, S.; Morton, J.; Shepherd, J.; Oh, Y.; Swope, K.; DeBeauchamp, J.L.; et al. Development of a SARS-CoV-2 Vaccine Candidate Using Plant-Based Manufacturing and a Tobacco Mosaic Virus-like Nano-Particle. Vaccines 2021, 9, 1347. [Google Scholar] [CrossRef] [PubMed]
- Jung, J.W.; Zahmanova, G.; Minkov, I.; Lomonossoff, G.P. Plant-based expression and characterization of SARS-CoV-2 virus-like particles presenting a native spike protein. Plant Biotechnol. J. 2022, 20, 1363–1372. [Google Scholar] [CrossRef]
- Peyret, H.; Steele, J.F.C.; Jung, J.-W.; Thuenemann, E.C.; Meshcheriakova, Y.; Lomonossoff, G.P. Producing Vaccines against Enveloped Viruses in Plants: Making the Impossible, Difficult. Vaccines 2021, 9, 780. [Google Scholar] [CrossRef]
- Moon, K.B.; Jeon, J.H.; Choi, H.; Park, J.S.; Park, S.J.; Lee, H.J.; Park, J.M.; Cho, H.S.; Moon, J.S.; Oh, H.; et al. Construction of SARS-CoV-2 virus-like particles in plant. Sci. Rep. 2022, 12, 1005. [Google Scholar] [CrossRef]
- Bachmann, M.F.; Jennings, G.T. Vaccine Delivery: A Matter of Size, Geometry, Kinetics and Molecular Patterns. Nat. Rev. Immunol. 2010, 10, 787–796. [Google Scholar] [CrossRef]
- Bradley, D.; Andjaparidze, A.; Cook, E.H.; McCaustland, K.; Balayan, M.; Stetler, H.; Velazquez, O.; Robertson, B.; Humphrey, C.; Kane, M.; et al. Aetiological Agent of Enterically Transmitted Non-A, Non-B Hepatitis. J. Gen. Virol. 1988, 69, 731–738. [Google Scholar] [CrossRef]
- Meng, J.; Dai, X.; Chang, J.C.; Lopareva, E.; Pillot, J.; Fields, H.A.; Khudyakov, Y.E. Identification and characterization of the neutralization epitope(s) of the hepatitis E virus. Virology 2001, 288, 203–211. [Google Scholar] [CrossRef]
- Emerson, S.U.; Clemente-Casares, P.; Moiduddin, N.; Arankalle, V.A.; Torian, U.; Purcell, R.H. Putative neutralization epitopes and broad cross-genotype neutralization of Hepatitis E virus confirmed by a quantitative cell-culture assay. J. Gen. Virol. 2006, 87, 697–704. [Google Scholar] [CrossRef] [PubMed]
- Jiménez de Oya, N.; Escribano-Romero, E.; Blázquez, A.-B.; Lorenzo, M.; Martín-Acebes, M.A.; Blasco, R.; Saiz, J.-C. Characterization of Hepatitis E Virus Recombinant ORF2 Proteins Expressed by Vaccinia Viruses. J. Virol. 2012, 86, 7880–7886. [Google Scholar] [CrossRef] [PubMed]
- Riddell, M.A.; Li, F.; Anderson, D.A. Identification of Immunodominant and Conformational Epitopes in the Capsid Protein of Hepatitis E Virus by Using Monoclonal Antibodies. J. Virol. 2000, 74, 8011–8017. [Google Scholar] [CrossRef] [PubMed]
- Zahmanova, G.G.; Mazalovska, M.; Takova, K.H.; Toneva, V.T.; Minkov, I.N.; Mardanova, E.S.; Ravin, N.V.; Lomonossoff, G.P. Rapid High-Yield Transient Expression of Swine Hepatitis E ORF2 Capsid Proteins in Nicotiana benthamiana Plants and Production of Chimeric Hepatitis E Virus-Like Particles Bearing the M2e Influenza Epitope. Plants 2020, 9, 29. [Google Scholar] [CrossRef]
- Mardanova, E.S.; Takova, K.H.; Toneva, V.T.; Zahmanova, G.G.; Tsybalova, L.M.; Ravin, N.V. A Plant-Based Transient Expression System for the Rapid Production of Highly Immunogenic Hepatitis E Virus-like Particles. Biotechnol. Lett. 2020, 42, 2441–2446. [Google Scholar] [CrossRef] [PubMed]
- Xing, L.; Kato, K.; Li, T.; Takeda, N.; Miyamura, T.; Hammar, L.; Cheng, R.H. Recombinant hepatitis E capsid protein self-assembles into a dual-domain T = 1 particle presenting native virus epitopes. Virology 1999, 265, 35–45. [Google Scholar] [CrossRef]
- Xing, L.; Li, T.C.; Miyazaki, N.; Simon, M.N.; Wall, J.S.; Moore, M.; Wang, C.Y.; Takeda, N.; Wakita, T.; Miyamura, T.; et al. Structure of hepatitis E virion-sized particle reveals an RNA-dependent viral assembly pathway. J. Biol. Chem. 2010, 285, 33175–33183. [Google Scholar] [CrossRef]
- Yamashita, T.; Mori, Y.; Miyazaki, N.; Cheng, R.H.; Yoshimura, M.; Unno, H.; Shima, R.; Moriishi, K.; Tsukihara, T.; Li, T.C.; et al. Biological and immunological characteristics of hepatitis E virus-like particles based on the crystal structure. Proc. Natl. Acad. Sci. USA 2009, 106, 12986–12991. [Google Scholar] [CrossRef]
- Guu, T.S.; Liu, Z.; Ye, Q.; Mata, D.A.; Li, K.; Yin, C.; Zhang, J.; Tao, Y.J. Structure of the hepatitis E virus-like particle suggests mechanisms for virus assembly and receptor binding. Proc. Natl. Acad. Sci. USA 2009, 106, 12992–12997. [Google Scholar] [CrossRef]
- Xing, L.; Wang, J.C.; Li, T.-C.; Yasutomi, Y.; Lara, J.; Khudyakov, Y.; Schofield, D.; Emerson, S.U.; Purcell, R.H.; Takeda, N.; et al. Spatial Configuration of Hepatitis E Virus Antigenic Domain. J. Virol. 2011, 85, 1117–1124. [Google Scholar] [CrossRef]
- Li, S.; Tang, S.; Seetharaman, J.; Yang, C.Y.; Gu, Y.; Zhang, J.; Du, H.; Shih, J.W.; Hew, C.L.; Sivaraman, J.; et al. Dimerization of hepatitis E virus capsid protein E2s domain is essential for virus-host interaction. PLoS Pathog. 2009, 5, e1000537. [Google Scholar] [CrossRef] [PubMed]
- Niikura, M.; Takamura, S.; Kim, G.; Kawai, S.; Saijo, M.; Morikawa, S.; Kurane, I.; Li, T.C.; Takeda, N.; Yasutomi, Y. Chimeric recombinant hepatitis E virus-like particles as an oral vaccine vehicle presenting foreign epitopes. Virology 2002, 293, 273–280. [Google Scholar] [CrossRef] [PubMed]
- Avdjieva, I.; Terziyski, I.; Zahmanova, G.; Simeonova, V.; Kulev, O.; Krustev, E.; Krachunov, M.; Nisheva, M.; Vassilev, D. Homology based computational modelling of hepatitis-E viral fusion capsid protein. C. R. l’Acad. Bulgare Des. Sci. 2019, 72, 358–364. [Google Scholar] [CrossRef]
- Jariyapong, P.; Xing, L.; van Houten, N.E.; Li, T.-C.; Weerachatyanukul, W.; Hsieh, B.; Moscoso, C.G.; Chen, C.-C.; Niikura, M.; Cheng, R.H. Chimeric Hepatitis E Virus-like Particle as a Carrier for Oral-Delivery. Vaccine 2013, 31, 417–424. [Google Scholar] [CrossRef]
- Biasini, M.; Bienert, S.; Waterhouse, A.; Arnold, K.; Studer, G.; Schmidt, T.; Kiefer, F.; Gallo Cassarino, T.; Bertoni, M.; Bordoli, L.; et al. SWISS-MODEL: Modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res. 2014, 42, 252–258. [Google Scholar] [CrossRef]
- Kiefer, F.; Arnold, K.; Kunzli, M.; Bordoli, L.; Schwede, T. The SWISS-MODEL Repository and associated resources. Nucleic Acids Res. 2009, 37, 387–392. [Google Scholar] [CrossRef]
- Arnold, K.; Bordoli, L.; Kopp, J.; Schwede, T. The SWISS-MODEL Workspace: A web-based environment for protein structure homology modeling. Bioinformatics 2006, 22, 195–201. [Google Scholar] [CrossRef]
- Guex, N.; Peitsch, M.; Schwede, T. Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: A historical perspective. Electrophoresis 2009, 30, 162–173. [Google Scholar] [CrossRef]
- Takova, K.; Koynarski, T.; Minkov, G.; Toneva, V.; Mardanova, E.; Ravin, N.; Lukov, G.L.; Zahmanova, G. Development and Optimization of an Enzyme Immunoassay to Detect Serum Antibodies against the Hepatitis E Virus in Pigs, Using Plant-Derived ORF2 Recombinant Protein. Vaccines 2021, 9, 991. [Google Scholar] [CrossRef]
- Zahmanova, G.; Mazalovska, M.; Takova, K.; Toneva, V.; Minkov, I.; Peyret, H.; Lomonossoff, G. Efficient Production of Chimeric Hepatitis B Virus-Like Particles Bearing an Epitope of Hepatitis E Virus Capsid by Transient Expression in Nicotiana benthamiana. Life 2021, 11, 64. [Google Scholar] [CrossRef]
- Nooraei, S.; Bahrulolum, H.; Hoseini, Z.S.; Katalani, C.; Hajizade, A.; Easton, A.J.; Ahmadian, G. Virus-like particles: Preparation, immunogenicity and their roles as nanovaccines and drug nanocarriers. J. Nanobiotechnol. 2021, 19, 59. [Google Scholar] [CrossRef] [PubMed]
- Chen, J.; Pompano, R.R.; Santiago, F.W.; Maillat, L.; Sciammas, R.; Sun, T.; Han, H.; Topham, D.J.; Chong, A.S.; Collier, J.H. The use of self-adjuvanting nanofiber vaccines to elicit high-affinity B cell responses to peptide antigens without inflammation. Biomaterials 2013, 34, 8776–8785. [Google Scholar] [CrossRef] [PubMed]
- Al-Halifa, S.; Gauthier, L.; Arpin, D.; Bourgault, S.; Archambault, D. Nanoparticlebased vaccines against respiratory viruses. Front. Immunol. 2019, 10, 22. [Google Scholar] [CrossRef] [PubMed]
- Larrue, H.; Abravanel, F.; Pe´ron, J.M. Hepatitis E, what’s the real issue? Liver Int. 2020, 40, 43–47. [Google Scholar] [CrossRef]
- Saunders, K.; Thuenemann, E.C.; Shah, S.N.; Peyret, H.; Kristianingsih, R.; Lopez, S.G.; Richardson, J.; Lomonossoff, G.P. The Use of a Replicating Virus Vector for in Planta Generation of Tobacco Mosaic Virus Nanorods Suitable for Metallization. Front. Bioeng. Biotechnol. 2022, 10, 877361. [Google Scholar] [CrossRef]
Method | HEV | HEV/M2e | HEV/RBD |
---|---|---|---|
Atomic force microscopy | particles (26 ± 5 nm) | particles (60 ± 7 nm) | particles (40 ± 4 nm) |
Electron microscopy | particles (18 ± 8 nm) | particles (42 ± 6 nm) | particles (33 ± 4 nm) |
Dynamic light scattering | particles (25 ± 4 nm) aggregates ~100 nm | particles (48 ± 3 nm) aggregates (200–300 nm) | particles (42 ± 2 nm) aggregates (200–300 nm) |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Mardanova, E.S.; Kotlyarov, R.Y.; Stuchinskaya, M.D.; Nikolaeva, L.I.; Zahmanova, G.; Ravin, N.V. 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. Int. J. Mol. Sci. 2022, 23, 15684. https://doi.org/10.3390/ijms232415684
Mardanova ES, Kotlyarov RY, Stuchinskaya MD, Nikolaeva LI, Zahmanova G, Ravin NV. 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. International Journal of Molecular Sciences. 2022; 23(24):15684. https://doi.org/10.3390/ijms232415684
Chicago/Turabian StyleMardanova, Eugenia S., Roman Y. Kotlyarov, Maya D. Stuchinskaya, Lyudmila I. Nikolaeva, Gergana Zahmanova, and Nikolai V. Ravin. 2022. "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" International Journal of Molecular Sciences 23, no. 24: 15684. https://doi.org/10.3390/ijms232415684
APA StyleMardanova, E. S., Kotlyarov, R. Y., Stuchinskaya, M. D., Nikolaeva, L. I., Zahmanova, G., & Ravin, N. V. (2022). 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. International Journal of Molecular Sciences, 23(24), 15684. https://doi.org/10.3390/ijms232415684