Archives of Virology

, Volume 164, Issue 2, pp 497–507 | Cite as

Development of sesbania mosaic virus nanoparticles for imaging

  • G. P. Vishnu Vardhan
  • M. HemaEmail author
  • C. Sushmitha
  • H. S. SavithriEmail author
  • Usha Natraj
  • M. R. N. Murthy
Original Article


The capsids of viruses have a high degree of symmetry. Therefore, virus nanoparticles (VNPs) can be programmed to display many imaging agents precisely. Plant VNPs are biocompatible, biodegradable and non-infectious to mammals. We have carried out bioconjugation of sesbania mosaic virus (SeMV), a well characterized plant virus, with fluorophores using reactive lysine-N-hydroxysuccinimide ester and cysteine-maleimide chemistries. Monitoring of cellular internalization of labelled SeMV nanoparticles (NPs) by confocal microscopy and flow cytometry showed that the particles have a natural preference for entry into MDA-MB-231 (breast cancer) cells, although they could also enter various other cell lines. The fluorescence of SeMV NPs labelled via the cysteines with Cy5.5 dye was found to be more stable and was detectable with greater sensitivity than that of particles labelled via the lysines with Alexa Fluor. Live-cell imaging using SeMV internally labelled with Cy5.5 showed that it could bind to MDA-MB-231 cells in less than 5 minutes and enter the cells within 15 minutes. The particles undergo endolysosomal degradation by 6 h as evidenced by their co-localization with LAMP-1. Far-western blot analysis with a HeLa cell membrane protein fraction showed that SeMV interacts with 54-, 35- and 33-kDa proteins, which were identified by mass spectrometry as vimentin, voltage-dependent anion-selective channel protein (VDAC1), and annexin A2 isoform 2 (ANXA2), respectively, suggesting that the particles may bind and enter the cell through these proteins. The results presented here demonstrate that the SeMV NPs provide a new platform technology that could be used to develop in vivo imaging and targeted drug delivery agents for cancer diagnosis and therapy.



This work was funded by Department of Biotechnology, India (BT/PR6711/NNT/28/622/2012). We also thank the Indian Institute of Science for financial support and for all the facilities. The Institute of Bioinformatics, Bengaluru, is gratefully acknowledged for help with mass spectrometric analysis. MRN and HSS thank the Department of Science and Technology (DST) for a J.C. Bose fellowship, and the Indian National Science Academy (INSA) for financial support. Vishnu Vardhan acknowledges DST-INSPIRE for providing a fellowship.

Supplementary material

705_2018_4097_MOESM1_ESM.doc (1 mb)
Supplementary material 1 (DOC 1060 kb)
705_2018_4097_MOESM2_ESM.avi (9.5 mb)
Supplementary material 2 (AVI 9730 kb)


  1. 1.
    Abraham A, Nataraj U, Karande AA, Gulati A, Murthy MRN, Murugesan S, Mukunda P, Savithri HS (2016) Intracellular delivery of antibodies by chimeric Sesbania mosaic virus (SeMV) virus like particles. Sci Rep 6:21803. CrossRefGoogle Scholar
  2. 2.
    Gulati A, Murthy A, Abraham A, Mohan K, Natraj U, Savithri HS, Murthy MRN (2016) Structural studies on chimeric Sesbania mosaic virus coat protein: revisiting SeMV assembly. Virology 489:34–43CrossRefGoogle Scholar
  3. 3.
    Bakshi A, Vishnu Vardhan GP, Hema M, Murthy MRN, Savithri HS (2017) Structural and functional characterization of sesbania mosaic virus. In: Mandal B, Rao G, Baranwal V, Jain R (eds) A century of plant virology in India. Springer, Singapore, pp 405–427CrossRefGoogle Scholar
  4. 4.
    Bhuvaneshwari M, Subramanya HS, Gopinath K, Savithri HS, Nayudu MV, Murthy MRN (1995) Structure of Sesbania mosaic virus at 3 Å resolution. Structure 3:1021–1030CrossRefGoogle Scholar
  5. 5.
    Chang JS, Chi SC (2015) GHSC70 is involved in the cellular entry of nervous necrosis virus. J Virol 89:61–70CrossRefGoogle Scholar
  6. 6.
    Culver JN, Brown AD, Zang F, Gnerlich M, Gerasopoulos K, Ghodssi R (2015) Plant virus directed fabrication of nanoscale materials and devices. Virology 480:200–212CrossRefGoogle Scholar
  7. 7.
    Czapar AE, Steinmetz NF (2017) Plant viruses and bacteriophages for drug delivery in medicine and biotechnology. Curr Opin Chem Biol 38:108–116CrossRefGoogle Scholar
  8. 8.
    Govind K, Makinen K, Savithri HS (2012) Sesbania mosaic virus (SeMV) infectious clone: possible mechanism of 3′ and 5′ end repair and role of polyprotein processing in viral replication. PLoS One 7:e31190CrossRefGoogle Scholar
  9. 9.
    Hema M, Bindi KP, Paul LC, He H, Neetu MG, Bradley LC, Reza AG, Sourabh S, Steinmetz NF (2017) Physalis mottle virus-like particles as nanocarriers for imaging reagents and drugs. Biomacromolecules 18:4141–4153CrossRefGoogle Scholar
  10. 10.
    Koudelka KJ, Destito G, Plummer EM, Trauger SA, Siuzdak G, Manchester M (2009) Endothelial targeting of cowpea mosaic virus (CPMV) via surface vimentin. PLoS Pathog 5:e1000417CrossRefGoogle Scholar
  11. 11.
    Koudelka KJ, Pitek AS, Manchester M, Steinmetz NF (2015) Virus-based nanoparticles as versatile nanomachines. Annu Rev Virol 2:379–401CrossRefGoogle Scholar
  12. 12.
    Lee KL, Hubbard LC, Hern S, Yildiz I, Gratzl M, Steinmetz NF (2013) Shape matters: the diffusion rates of TMV rods and CPMV icosahedrons in a spheroid model of extracellular matrix are distinct. Biomater Sci 1:10CrossRefGoogle Scholar
  13. 13.
    Lokesh GL, Gopinath K, Sathesh Kumar PS, Savithri HS (2001) Complete nucleotide sequence of Sesbania mosaic virus: a new virus species of the genus Sobemovirus. Arch Virol 146:209–223CrossRefGoogle Scholar
  14. 14.
    Ma Y, Nolte RJ, Cornelissen JJ (2012) Virus-based nanocarriers for drug delivery. Adv Drug Deliv Rev 64:811–825CrossRefGoogle Scholar
  15. 15.
    Mateu MG (2011) Virus engineering: functionalization and stabilization. Protein Eng Des Sel 24:53–63CrossRefGoogle Scholar
  16. 16.
    Narayanan KB, Han SS (2017) Icosahedral plant viral nanoparticles-bioinspired synthesis of nanomaterials/nanostructures. Adv Colloid Interface Sci 248:1–19CrossRefGoogle Scholar
  17. 17.
    Narayanan KB, Han SS (2017) Helical plant viral nanoparticles-bioinspired synthesis of nanomaterials and nanostructures. Bioinspir Biomim 12:031001CrossRefGoogle Scholar
  18. 18.
    Plummer EM, Manchester M (2013) Endocytic uptake pathways utilized by CPMV nanoparticles. Mol Pharm 10:26–32CrossRefGoogle Scholar
  19. 19.
    Pokorski JK, Steinmetz NF (2011) The art of engineering viral nanoparticles. Mol Pharm 8:29–43CrossRefGoogle Scholar
  20. 20.
    Sathesh Kumar PS, Lokesh GL, Murthy MRN, Savithri HS (2005) The role of arginine-rich motif and β-annulus in the assembly and stability of Sesbania mosaic virus capsids. J Mol Biol 35:3447–3458Google Scholar
  21. 21.
    Vishnu Vardhan GP, Savithri HS, Murthy MRN, Hema M (2016) Biodistribution and toxicity evaluation of Sesbania mosaic virus nanoparticles in mice. Arch Virol 161:2673–2681CrossRefGoogle Scholar
  22. 22.
    Wen AM, Cho CF, Lewis JD, Steinmetz NF (2015) The application of plant viral nanoparticles in tissue-specific imaging. In: Mikhail YB (ed) Nanotechnology for biomedical imaging and diagnostics: from nanoparticle design to clinical applications. Wiley, New York, pp 401–427Google Scholar
  23. 23.
    Wu Y, Li Q, Chen Z (2007) Detecting protein-protein interactions by far western blotting. Nat Protoc 2:3278–3284CrossRefGoogle Scholar
  24. 24.
    Young M, Willits D, Uchida M, Douglas T (2008) Plant viruses as biotemplates for materials and their use in nanotechnology. Annu Rev Phytopathol 46:361–384CrossRefGoogle Scholar
  25. 25.
    Zhang S, Yu M, Guo Q, Li R, Li G, Tan S, Li X, Wei Y, Wu M (2015) Annexin A2 binds to endosomes and negatively regulated TLR4-triggered inflammatory responses via the TRAM-TRIF pathway. Sci Rep 5:15859CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of BiochemistryIndian Institute of ScienceBengaluruIndia
  2. 2.Molecular Biophysics UnitIndian Institute of ScienceBengaluruIndia
  3. 3.Department of VirologySri Venkateswara UniversityTirupatiIndia

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