Development of sesbania mosaic virus nanoparticles for imaging
Autor: | Usha Natraj, C. Sushmitha, G. P. Vishnu Vardhan, H.S. Savithri, M. Hema, Mathur R. N. Murthy |
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Rok vydání: | 2018 |
Předmět: |
Molecular Biophysics Unit
Biology Biochemistry Fluorescence Plant Viruses law.invention Flow cytometry Cell membrane HeLa 03 medical and health sciences Confocal microscopy law Cell Line Tumor Virology medicine Humans Fluorescent Dyes 030304 developmental biology Alexa Fluor 0303 health sciences Microscopy Confocal Bioconjugation medicine.diagnostic_test 030306 microbiology General Medicine Flow Cytometry biology.organism_classification Molecular Imaging Blot medicine.anatomical_structure Targeted drug delivery Biophysics Nanoparticles Lysosomes |
Zdroj: | Archives of Virology. 164:497-507 |
ISSN: | 1432-8798 0304-8608 |
Popis: | 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. |
Databáze: | OpenAIRE |
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