Potato Virus X as a Novel Platform for Potential Biomedical Applications

Steinmetz, Nicole F.; Mertens, Marianne E.; Taurog, Rebecca E.; Johnson, John E.; Commandeur, Ulrich; Fischer, Rainer; Manchester, Marianne

doi:10.1021/nl9035753

Cited by 100 publications

(108 citation statements)

References 44 publications

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“…14,17,18,21 The characterization methods described are valuable for ensuring particle integrity and determining extent of conjugation. Molar excesses can be adjusted accordingly depending on the application and the degree of functionalization desired.…”

Section: Discussionmentioning

confidence: 99%

Viral Nanoparticles for <em>In vivo</em> Tumor Imaging

Wen

Lee

Yildiz

et al. 2012

JoVE

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The use of nanomaterials has the potential to revolutionize materials science and medicine. Currently, a number of different nanoparticles are being investigated for applications in imaging and therapy. Viral nanoparticles (VNPs) derived from plants can be regarded as self-assembled bionanomaterials with defined sizes and shapes. Plant viruses under investigation in the Steinmetz lab include icosahedral particles formed by Cowpea mosaic virus (CPMV) and Brome mosaic virus (BMV), both of which are 30 nm in diameter. We are also developing rod-shaped and filamentous structures derived from the following plant viruses: Tobacco mosaic virus (TMV), which forms rigid rods with dimensions of 300 nm by 18 nm, and Potato virus X (PVX), which form filamentous particles 515 nm in length and 13 nm in width (the reader is referred to refs. 1 and 2 for further information on VNPs).From a materials scientist's point of view, VNPs are attractive building blocks for several reasons: the particles are monodisperse, can be produced with ease on large scale in planta, are exceptionally stable, and biocompatible. Also, VNPs are "programmable" units, which can be specifically engineered using genetic modification or chemical bioconjugation methods 3. The structure of VNPs is known to atomic resolution, and modifications can be carried out with spatial precision at the atomic level 4 , a level of control that cannot be achieved using synthetic nanomaterials with current state-of-the-art technologies.In this paper, we describe the propagation of CPMV, PVX, TMV, and BMV in Vigna ungiuculata and Nicotiana benthamiana plants. Extraction and purification protocols for each VNP are given. Methods for characterization of purified and chemically-labeled VNPs are described. In this study, we focus on chemical labeling of VNPs with fluorophores (e.g. Alexa Fluor 647) and polyethylene glycol (PEG). The dyes facilitate tracking and detection of the VNPs [5][6][7][8][9][10] , and PEG reduces immunogenicity of the proteinaceous nanoparticles while enhancing their pharmacokinetics 8,11 . We demonstrate tumor homing of PEGylated VNPs using a mouse xenograft tumor model. A combination of fluorescence imaging of tissues ex vivo using Maestro Imaging System, fluorescence quantification in homogenized tissues, and confocal microscopy is used to study biodistribution. VNPs are cleared via the reticuloendothelial system (RES); tumor homing is achieved passively via the enhanced permeability and retention (EPR) effect 12 . The VNP nanotechnology is a powerful plug-and-play technology to image and treat sites of disease in vivo. We are further developing VNPs to carry drug cargos and clinically-relevant imaging moieties, as well as tissue-specific ligands to target molecular receptors overexpressed in cancer and cardiovascular disease. Video LinkThe video component of this article can be found at

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Section: Discussionmentioning

confidence: 99%

Viral Nanoparticles for <em>In vivo</em> Tumor Imaging

Wen

Lee

Yildiz

et al. 2012

JoVE

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“…Filamentous particles also present ligand in a more efficient manner. A rod-shaped particle may in theory interact with a larger number of binding sites on the cell surface, thus increasing targeting sensitivity and specificity [10].…”

Section: Introductionmentioning

confidence: 99%

A new application of plant virus nanoparticles as drug delivery in breast cancer

et al. 2015

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Nanoparticles based on non-pathogenic viruses have opened up a novel sector in nanotechnology. Viral nanoparticles based on plant viruses have clear advantages over any synthetic nanoparticles as they are biocompatible and biodegradable self-assembled and can be produced inexpensively on a large scale. From several such under-development platforms, only a few have been characterized in the target-specific drugs into the cells. Potato virus X is presented as a carrier of the chemotherapeutic drug Herceptin that is currently used as a targeted therapy in (HER2+) breast cancer patients. Here, we used nanoparticles formed from the potato virus X to conjugate the Herceptin (Trastuzumab) monoclonal antibody as a new option in specific targeting of breast cancer. Bioconjugation was performed by EDC/sulfo-N-hydroxysuccinimide (sulfo-NHS) in a two-step protocol. Then, the efficiency of conjugation was investigated by different methods, including sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, ELISA, Zetasizer, and transmission electron microscopy. SDS-PAGE and Western blot analysis confirmed an 82-kDa protein band that resulted from conjugation of potato virus X (PVX) coat protein (27 kDa) to heavy chain of Herceptin (55 kDa). Zeta potential values for conjugated particles, PVX, and HER were -7.05, -21.4, and -1.48, respectively. We investigated the efficiency of PVX-Herceptin to induce SK-OV-3 and SK-BR-3 cells (HER2 positive cell lines) apoptosis. We therefore counted cells and measured apoptosis by flow cytometry assay, then compared with Herceptin alone. Based on our data, we confirmed the conjugation of PVX and Herceptin. This study suggests that the PVX-Herceptin conjugates enable Herceptin to become more potential therapeutic tools.

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“…For example, Fisher and Manchester used these conditions for functionalization of the rod-shaped potato virus X with fluorescent dyes (159). Wang and coworkers chemoselectively functionalized the surface of horse spleen apoferritin, a cage-like protein structure composed of 24 structurally equivalent subunits (160).…”

Section: Bionanoparticlesmentioning

confidence: 99%

Click Chemistry for Drug Delivery Nanosystems

et al. 2011

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The purpose of this Expert Review is to discuss the impact of click chemistry in nanosized drug delivery systems. Since the introduction of the click concept by Sharpless and coworkers in 2001, numerous examples of click reactions have been reported for the preparation and functionalization of polymeric micelles and nanoparticles, liposomes and polymersomes, capsules, microspheres, metal and silica nanoparticles, carbon nanotubes and fullerenes, or bionanoparticles. Among these click processes, Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) has attracted most attention based on its high orthogonality, reliability, and experimental simplicity for non-specialists. A renewed interest in the use of efficient classical transformations has been also observed (e.g., thiol-ene coupling, Michael addition, Diels-Alder). Special emphasis is also devoted to critically discuss the click concept, as well as practical aspects of application of CuAAC to ensure efficient and harmless bioconjugation.

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Potato Virus X as a Novel Platform for Potential Biomedical Applications

Cited by 100 publications

References 44 publications

Viral Nanoparticles for <em>In vivo</em> Tumor Imaging

Viral Nanoparticles for <em>In vivo</em> Tumor Imaging

A new application of plant virus nanoparticles as drug delivery in breast cancer

Click Chemistry for Drug Delivery Nanosystems

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