Abstract
Senecavirus A (SVA) is a picornavirus that causes vesicular disease in swine, and the inactivated vaccine is used to prevent and control SVA infection. To develop a new chromatography strategy for the purification and concentration of SVA vaccine antigens, we inserted a 6×His-tag at the VP1 C-terminal of the SVA/HLJ/CHA/2016 in an infectious clone to rescue a His-tagged SVA. The constructed and rescued recombinant virus, named as rSVA-His, exhibited similar growth kinetics to that of its parental virus. In addition, the expression of a 6×His-tag on the surface of SVA showed genetic stability in cell passages in vitro, which allowed one-step purification of SVA antigens by Ni2+ affinity columns. Furthermore, the immunogenicity of the inactivated rSVA-His was evaluated by inoculating rabbits and detecting neutralizing antibodies. The animals receiving two doses of the inactivated rSVA-His emulsified with oil adjuvant developed a high titer of neutralizing antibodies, indicating that SVA VP1 is tolerant to His-tag insertion without detriment to its antigenicity. In summary, the constructed 6×His-tagged SVA may offer a feasible approach to the affinity purification and concentration of antigens in the process of SVA inactivated vaccine production.
Highlights
Senecavirus A (SVA), formerly named Seneca Valley Virus (SVV), is one of the causative agents of vesicular diseases in swine
SVA is the single member of the genus Senecavirus within the family Picornaviridae and shares some common features with other picornaviruses: (1) a non-enveloped virus; (2) a single-stranded positive-sense RNA genome flanked by an internal ribosomal entry site at the 5 end and a polyadenylated tail at the 3 end; (3) a large, single open reading frame encoding a polyprotein; (4) a translated polyprotein precursor with the standard L-4-3-4 layout being processed into mature proteins by virus-encoded proteinases [2]
The result showed that the rSVA-His in the infected cells could be detected by both anti-His monoclonal antibody (mAb) and anti-SVA VP2 mAb, while the anti-His mAb did not react to the SVA-WT in the infected cells (Figure 2)
Summary
Senecavirus A (SVA), formerly named Seneca Valley Virus (SVV), is one of the causative agents of vesicular diseases in swine. SVA is the single member of the genus Senecavirus within the family Picornaviridae and shares some common features with other picornaviruses: (1) a non-enveloped virus; (2) a single-stranded positive-sense RNA genome flanked by an internal ribosomal entry site at the 5 end and a polyadenylated tail at the 3 end; (3) a large, single open reading frame encoding a polyprotein; (4) a translated polyprotein precursor with the standard L-4-3-4 layout being processed into mature proteins by virus-encoded proteinases [2] These mature proteins include four structural proteins (VP1—VP4) and eight nonstructural proteins (L, 2A-2B-2C-3A-3B-3C-3D) [2]. It was reported that SVA is linked to increased mortality in neonatal piglets with high morbidity [18]
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