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Freeze-Drying To Produce Efficacious CPMV Virus-like Particles.

ABSTRACT: In situ cancer vaccination that uses immune stimulating agents is revolutionizing the way that cancer is treated. In this realm, viruses and noninfectious virus-like particles have gained significant traction in reprogramming the immune system to recognize and eliminate malignancies. Recently, cowpea mosaic virus-like particles (VLPs) have shown exceptional promise in their ability to fight a variety of cancers. However, the current methods used to produce CPMV VLPs rely on agroinfiltration in plants. These protocols remain complicated and labor intensive and have the potential to introduce unwanted immunostimulatory agents, like lipopolysaccharides. This Letter describes a simple "post-processing" method to remove RNA from wild-type CPMV, while retaining the structure and function of the capsid. Lyophilization was able to eject encapsulated RNA to form lyo-eCPMV and, when purified, eliminated nearly all traces of encapsulated RNA. Lyo-eCPMV was characterized by cryo-electron microscopy single particle reconstruction to confirm the structural integrity of the viral capsid. Finally, lyo-eCPMV showed equivalent anticancer efficacy as eCPMV, produced by agroinfiltration, when using an invasive melanoma model. These results describe a straightforward method to prepare CPMV VLPs from infectious virions.

SUBMITTER: Zheng Y

PROVIDER: S-EPMC7272238 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature

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Freeze-Drying To Produce Efficacious CPMV Virus-like Particles.

Zheng Yi Y Lee Parker W PW Wang Chao C Thomas Linda D LD Stewart Phoebe L PL Steinmetz Nicole F NF Pokorski Jonathan K JK

Nano letters 20190225 3

In situ cancer vaccination that uses immune stimulating agents is revolutionizing the way that cancer is treated. In this realm, viruses and noninfectious virus-like particles have gained significant traction in reprogramming the immune system to recognize and eliminate malignancies. Recently, cowpea mosaic virus-like particles (VLPs) have shown exceptional promise in their ability to fight a variety of cancers. However, the current methods used to produce CPMV VLPs rely on agroinfiltration in p ...[more]

PMID: 30801195

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Project description:Vacuum freeze-drying of biological materials is one of the best methods of water removal, with final products of highest quality. The solid state of water during freeze-drying protects the primary structure and the shape of the products with minimal volume reduction. In addition, the lower temperatures in the process allow maximal nutrient and bioactive compound retention. This technique has been successfully applied to diverse biological materials, such as meats, coffee, juices, dairy products, cells, and bacteria, and is standard practice for penicillin, hormones, blood plasma, vitamin preparations, etc. Despite its many advantages, having four to ten times more energy requirements than regular hot air drying, freeze-drying has always been recognized as the most expensive process for manufacturing a dehydrated product. The application of the freeze-drying process to plant-based foods has been traditionally dedicated to the production of space shuttle goods, military or extreme-sport foodstuffs, and specialty foods such as coffee or spices. Recently, the market for 'natural' and 'organic' products is, however, strongly growing as well as the consumer's demand for foods with minimal processing and high quality. From this perspective, the market for freeze-dried plant-based foods is not only increasing but also diversifying. Freeze-dried fruits and vegetables chunks, pieces, or slices are nowadays majorly used in a wide range of food products such as confectionaries, morning cereals, soups, bakeries, meal boxes, etc. Instant drinks are prepared out of freeze-dried tea, coffee, or even from maple syrup enriched with polyphenol concentrated extracts from trees. The possibilities are endless. In this review, the application of freeze-drying to transform plant-based foods was analyzed, based on the recent research publications on the subject and personal unpublished data. The review is structured around the following related topics: latest applications of freeze-drying to plant-based foods, specific technological problems that could be found when freeze-drying such products (i.e., presence of cuticle; high sugar or lipid concentration), pretreatments and intensification technologies employed in freeze-drying of plant-based foods, and quality issues of these freeze-dried products.

Dataset Information

Freeze-Drying To Produce Efficacious CPMV Virus-like Particles.

Publications

Freeze-Drying To Produce Efficacious CPMV Virus-like Particles.

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