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Nanomanufacturing of Tobacco Mosaic Virus-Based Spherical Biomaterials Using a Continuous Flow Method.

ABSTRACT: Nanomanufacturing of nanoparticles is critical for potential translation and commercialization. Continuous flow devices can alleviate this need through unceasing production of nanoparticles. Here we demonstrate the scaled-up production of spherical nanoparticles functionalized with biomedical cargos from the rod-shaped plant virus tobacco mosaic virus (TMV) using a mesofluidic, continued flow method. Production yields were increased 30-fold comparing the mesofluidic device versus batch methods. Finally, we produced MRI contrast agents of select sizes, with per particle relaxivity reaching 979,218 mM^-1 s^-1 at 60 MHz. These TMV-based spherical nanoparticle MRI contrast agents are in the top echelon of relaxivity per nanoparticle.

SUBMITTER: Bruckman MA

PROVIDER: S-EPMC4426350 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

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Nanomanufacturing of Tobacco Mosaic Virus-Based Spherical Biomaterials Using a Continuous Flow Method.

Bruckman Michael A MA VanMeter Allen A Steinmetz Nicole F NF

ACS biomaterials science & engineering 20141209 1

Nanomanufacturing of nanoparticles is critical for potential translation and commercialization. Continuous flow devices can alleviate this need through unceasing production of nanoparticles. Here we demonstrate the scaled-up production of spherical nanoparticles functionalized with biomedical cargos from the rod-shaped plant virus tobacco mosaic virus (TMV) using a mesofluidic, continued flow method. Production yields were increased 30-fold comparing the mesofluidic device versus batch methods. ...[more]

PMID: 25984569

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Project description:BACKGROUND: Although tobacco mosaic virus (TMV) coat protein (CP) has been isolated from virus particles and its crystals have grown in ammonium sulfate buffers for many years, to date, no one has reported on the crystallization of recombinant TMV-CP connecting peptides expressed in E. coli. METHODS: In the present papers genetically engineered TMV-CP was expressed, into which hexahistidine (His) tags or glutathione-S-transferase (GST) tags were incorporated. Considering that GST-tags are long peptides and His-tags are short peptides, an attempt was made to grow crystals of TMV-CP cleaved GST-tags (WT-TMV-CP32) and TMV-CP incorporated His-tags (WT-His-TMV-CP12) simultaneously in ammonium sulfate buffers and commercial crystallization reagents. It was found that the 20S disk form of WT-TMV-CP32 and WT-His-TMV-CP12 did not form high resolution crystals by using various crystallization buffers and commercial crystallization reagents. Subsequently, a new experimental method was adopted in which a range of truncated TMV-CP was constructed by removing several amino acids from the N- or the C-terminal, and high resolution crystals were grown in ammonium sulfate buffers and commercial crystallization reagents. RESULTS: The new crystallization method was developed and 3.0 Å resolution macromolecular crystal was thereby obtained by removing four amino acids at the C-terminal of His-TMV-CP and connecting six His-tags at the N-terminal of His-TMV-CP (TR-His-TMV-CP19). The Four-layer aggregate disk structure of TR-His-TMV-CP19 was solved. This phenomenon showed that peptides at the C-terminus hindered the growth of high resolution crystals and the peptides interactions at the N-terminus were attributed to the quality of TMV-CP crystals. CONCLUSION: A 3.0 Å resolution macromolecular crystal of TR-His-TMV-CP19 was obtained and the corresponding structure was solved by removing four amino acids at the C-terminus of TMV-CP and connecting His-tags at the N-terminus of TMV-CP. It indicated that short peptides influenced the resolution of TMV-CP crystals.

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Nanomanufacturing of Tobacco Mosaic Virus-Based Spherical Biomaterials Using a Continuous Flow Method.

Publications

Nanomanufacturing of Tobacco Mosaic Virus-Based Spherical Biomaterials Using a Continuous Flow Method.

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