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      Cell-free protein synthesis of norovirus virus-like particles

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          Abstract

          Cell-free protein expression of norovirus virus-like-particles.

          Abstract

          Norovirus vaccine development largely depends on recombinant virus-like-particles (VLPs). Norovirus VLPs have been produced in several cell-based expression systems with long production times. Here we report, for the first time, that norovirus VLPs can be expressed and assembled by using a cell-free protein expression system within four hours.

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          Replication of human noroviruses in stem cell-derived human enteroids

          Khalil Ettayebi, Sue Crawford, Kosuke Murakami … (2016)
          The major barrier to research and development of effective interventions for human noroviruses (HuNoVs) has been the lack of a robust and reproducible in vitro cultivation system. HuNoVs are the leading cause of gastroenteritis worldwide. We report the successful cultivation of multiple HuNoV strains in enterocytes in stem cell-derived, nontransformed human intestinal enteroid monolayer cultures. Bile, a critical factor of the intestinal milieu, is required for strain-dependent HuNoV replication. Lack of appropriate histoblood group antigen expression in intestinal cells restricts virus replication, and infectivity is abrogated by inactivation (e.g., irradiation, heating) and serum neutralization. This culture system recapitulates the human intestinal epithelium, permits human host-pathogen studies of previously noncultivatable pathogens, and allows the assessment of methods to prevent and treat HuNoV infections.
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            Cell-free protein synthesis: applications come of age.

            Erik Carlson, Rui Gan, C Hodgman … (2012)
            Cell-free protein synthesis has emerged as a powerful technology platform to help satisfy the growing demand for simple and efficient protein production. While used for decades as a foundational research tool for understanding transcription and translation, recent advances have made possible cost-effective microscale to manufacturing scale synthesis of complex proteins. Protein yields exceed grams protein produced per liter reaction volume, batch reactions last for multiple hours, costs have been reduced orders of magnitude, and reaction scale has reached the 100-liter milestone. These advances have inspired new applications in the synthesis of protein libraries for functional genomics and structural biology, the production of personalized medicines, and the expression of virus-like particles, among others. In the coming years, cell-free protein synthesis promises new industrial processes where short protein production timelines are crucial as well as innovative approaches to a wide range of applications. Copyright © 2011 Elsevier Inc. All rights reserved.
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              Portable, On-Demand Biomolecular Manufacturing.

              Keith Pardee, Shimyn Slomovic, Peter Nguyen … (2016)
              Synthetic biology uses living cells as molecular foundries for the biosynthesis of drugs, therapeutic proteins, and other commodities. However, the need for specialized equipment and refrigeration for production and distribution poses a challenge for the delivery of these technologies to the field and to low-resource areas. Here, we present a portable platform that provides the means for on-site, on-demand manufacturing of therapeutics and biomolecules. This flexible system is based on reaction pellets composed of freeze-dried, cell-free transcription and translation machinery, which can be easily hydrated and utilized for biosynthesis through the addition of DNA encoding the desired output. We demonstrate this approach with the manufacture and functional validation of antimicrobial peptides and vaccines and present combinatorial methods for the production of antibody conjugates and small molecules. This synthetic biology platform resolves important practical limitations in the production and distribution of therapeutics and molecular tools, both to the developed and developing world.
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                Author and article information

                Contributors
                Journal
                Journal ID (publisher-id): RSCACL
                Title: RSC Advances
                Abbreviated Title: RSC Adv.
                Publisher: Royal Society of Chemistry (RSC)
                ISSN (Electronic): 2046-2069
                Publication date Created: 2017
                Publication date (Electronic): 2017
                Volume: 7
                Issue: 46
                Pages: 28837-28840
                Affiliations
                [1 ]Department of Biological Systems Engineering
                [2 ]Virginia Polytechnic Institute and State University
                [3 ]Blacksburg
                [4 ]USA
                [5 ]Department of Biomedical Sciences and Pathobiology
                Article
                DOI: 10.1039/C7RA03742B
                SO-VID: 75074418-d201-42e7-9532-8cfe22e64e78
                Copyright © © 2017
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