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

Francesca Bugli, Massimiliano Papi, Brunella Posteraro, Maurizio Sanguinetti, Alessandro Arcovito, Margherita Cacaci, Cecilia Martini, Francesco Paroni Sterbini, Riccardo Torelli, Valentina Palmieri, Bruno Giardina, Valeria Caprettini, Stefano Della Longa

Risultato della ricerca: Contributo in rivistaArticolo in rivistapeer review

21 Citazioni (Scopus)

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 microns 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, and also represents a top-quality biomaterial to be further modified for biotechnological purposes.
Lingua originaleEnglish
pagine (da-a)2727-2739
Numero di pagine13
RivistaInternational Journal of Nanomedicine
Volume9
DOI
Stato di pubblicazionePubblicato - 2014

Keywords

  • SUMO fusion tag
  • Virus-like particles
  • human rotavirus vaccine
  • protein-based nanotubes

Fingerprint Entra nei temi di ricerca di 'Synthesis and characterization of different immunogenic viral nanoconstructs from rotavirus VP6 inner capsid protein'. Insieme formano una fingerprint unica.

Cita questo