Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
| Autor: | Maurizio Sanguinetti, Riccardo Torelli, Francesca Bugli, Francesco Paroni Sterbini, Brunella Posteraro, Valentina Palmieri, Cecilia Martini, Stefano Della Longa, Valeria Caprettini, Massimiliano Papi, Margherita Cacaci, Alessandro Arcovito, Bruno Giardina |
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| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
SUMO fusion tag
viruses Biophysics Pharmaceutical Science Bioengineering Biology medicine.disease_cause Protein Engineering Inclusion bodies Rotavirus Infections Biomaterials Mice International Journal of Nanomedicine Rotavirus Drug Discovery medicine Animals Settore BIO/10 - BIOCHIMICA Escherichia coli Antigens Viral Biological Products Mice Inbred BALB C Vaccines Synthetic Immunogenicity Capsomere Organic Chemistry Methodology virus diseases Protein engineering General Medicine Virus-like particles Molecular biology human rotavirus vaccine Genetic Enhancement Treatment Outcome Capsid Nanoparticles Capsid Proteins Female Thioredoxin Settore BIO/19 - MICROBIOLOGIA GENERALE protein-based nanotubes |
| Zdroj: | International Journal of Nanomedicine |
| ISSN: | 1178-2013 |
| Popis: | Francesca Bugli,1 Valeria Caprettini,2 Margherita Cacaci,1 Cecilia Martini,1 Francesco Paroni Sterbini,1 Riccardo Torelli,1 Stefano Della Longa,3 Massimiliano Papi,4 Valentina Palmieri,4 Bruno Giardina,5 Brunella Posteraro,1 Maurizio Sanguinetti,1 Alessandro Arcovito5 1Istituto di Microbiologia, Università Cattolica del Sacro Cuore, 2Dipartimento di Fisica, Sapienza Università di Roma, Rome, 3Dipartimento di Medicina Clinica, Sanità Pubblica, Scienze della Vita e dell’Ambiente, Università dell’Aquila, L’Aquila, 4Istituto di Fisica, 5Istituto di Biochimica e Biochimica Clinica, Università Cattolica del Sacro Cuore, Rome, Italy Abstract: In order to deliver low-cost viral capsomeres from a large amount of soluble viral VP6protein from human rotavirus, we developed and optimized a biotechnological platform in Escherichia coli. Specifically, three different expression protocols were compared, differing in their genetic constructs, ie, a simple native histidine-tagged VP6sequence, VP6fused to thioredoxin, and VP6obtained with the newly described small ubiquitin-like modifier (SUMO) fusion system. Our results demonstrate that the histidine-tagged protein does not escape the accumulation in the inclusion bodies, and that SUMO is largely superior to the thioredoxin-fusion tag in enhancing the expression and solubility of VP6protein. Moreover, the VP6protein produced according to the SUMO fusion tag displays well-known assembly properties, as observed in both transmission electron microscopy and atomic force microscopy images, giving rise to either VP6trimers,60nm spherical virus-like particles, or nanotubes a few micron long. This different quaternary organization of VP6shows a higher level of immunogenicity for the elongated structures with respect to the spheres or the protein trimers. Therefore, the expression and purification strategy presented here – providing a large amount of the viral capsid protein in the native form with relatively simple, rapid, and economical procedures – opens a new route toward large-scale production of a more efficient antigenic compound to be used as a vaccination tool or as an adjuvant, as well as represents a top-quality biomaterial to be further modified for biotechnological purposes. Keywords: Virus-like particles, protein-based nanotubes, SUMO fusion tag, human rotavirus vaccine |
| Databáze: | OpenAIRE |
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