Production and Easy One-Step Purification of Bluetongue Recombinant VP7 from Infected Sf9 Supernatant for an Immunoenzymatic Assay (ELISA)

• DOI: 10.1007/s12033-020-00282-8
• Publication Date: 2020-10-19T17:03:03Z
• Authors: S. Ulisse; M. Iorio; G. Armillotta; C. Laguardia; L. Testa; S. Capista; P. Centorame; S. Traini; A. Serroni; F. Monaco; M. Caporale; M. T. Mercante; M. Di Ventura
• Keywords: Sf9; Orbivirus; Culicoides; Recombinant virus

Bluetongue (BT) is non-contagious, vector-borne viral disease of domestic and wild ruminants, transmitted by midges (Culicoides spp.) caused virus (BTV). BTV the type species Orbivirus genus within Reoviridae family possesses a genome consisting 10 double-stranded RNA segments encoding 7 structural 4 nonstructural proteins. Viral Protein (VP7) major sera group-specific protein good antigen candidate for immunoenzymatic assays BT diagnosis. In our work, BTV-2 recombinant VP7 (BTV-2 recVP7), expressed in Spodoptera frugiperda (Sf9) cells using baculovirus system, was produced purified affinity chromatography from supernatant infected cell culture. The use allowed us to obtain high quantity with purity level an easy one-step procedure, rather than multistep purification pellet. RecVP7-BTV2 detected MAb anti-BTV Western blot it used develop assay.