Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein

Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein

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, U...

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Autores principales: Bugli F, Caprettini V, Cacaci M, Martini C, Paroni Sterbini F, Torelli R, Della Longa S, Papi M, Palmieri V, Giardina B, Posteraro B, Sanguinetti M, Arcovito A
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2014
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Medicine (General)
R5-920
Acceso en línea:https://doaj.org/article/084d374288dc48f199cac5ab4aaf98c7
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spelling oai:doaj.org-article:084d374288dc48f199cac5ab4aaf98c72021-12-02T02:21:19ZSynthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein1178-2013https://doaj.org/article/084d374288dc48f199cac5ab4aaf98c72014-05-01T00:00:00Zhttp://www.dovepress.com/synthesis-and-characterization-of-different-immunogenic-viral-nanocons-a17072https://doaj.org/toc/1178-2013 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 VP6 protein 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 VP6 sequence, VP6 fused to thioredoxin, and VP6 obtained 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 VP6 protein. Moreover, the VP6 protein 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 VP6 trimers, 60 nm spherical virus-like particles, or nanotubes a few micron long. This different quaternary organization of VP6 shows 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 vaccineBugli FCaprettini VCacaci MMartini CParoni Sterbini FTorelli RDella Longa SPapi MPalmieri VGiardina BPosteraro BSanguinetti MArcovito ADove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2014, Iss Issue 1, Pp 2727-2739 (2014)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
Bugli F
Caprettini V
Cacaci M
Martini C
Paroni Sterbini F
Torelli R
Della Longa S
Papi M
Palmieri V
Giardina B
Posteraro B
Sanguinetti M
Arcovito A
Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
description 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 VP6 protein 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 VP6 sequence, VP6 fused to thioredoxin, and VP6 obtained 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 VP6 protein. Moreover, the VP6 protein 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 VP6 trimers, 60 nm spherical virus-like particles, or nanotubes a few micron long. This different quaternary organization of VP6 shows 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
format article
author Bugli F
Caprettini V
Cacaci M
Martini C
Paroni Sterbini F
Torelli R
Della Longa S
Papi M
Palmieri V
Giardina B
Posteraro B
Sanguinetti M
Arcovito A
author_facet Bugli F
Caprettini V
Cacaci M
Martini C
Paroni Sterbini F
Torelli R
Della Longa S
Papi M
Palmieri V
Giardina B
Posteraro B
Sanguinetti M
Arcovito A
author_sort Bugli F
title Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
title_short Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
title_full Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
title_fullStr Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
title_full_unstemmed Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein
title_sort synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus vp6 inner capsid protein
publisher Dove Medical Press
publishDate 2014
url https://doaj.org/article/084d374288dc48f199cac5ab4aaf98c7
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