Recombinant occlusion bodies of baculovirus as carriers of a nonstructural protein of foot-and-mouth disease virus

Authors
Diez, Michay; Trotta, Myrian Vanesa; Alfonso, Victoria; Taboga, Oscar Alberto; Lopez, Maria Gabriela
Publication Year
2018
Language
English
Format
article
Status
Published version
Description
Here, we developed a diagnostic ELISA for foot-and-mouth disease using recombinant occlusion bodies (rOBs) of baculovirus. We fused Δ3AB1–3, a polypeptide derived from non-structural proteins of foot-and-mouth disease virus, to polyhedrin (POLH), the major constituent of OBs, under polh promoter. To further assess the most convenient strategy to improve yields, we designed two recombinant baculoviruses, vPOLH and vPOLHE44G. These carried the sequence of the fusion protein POLH-Δ3AB1–3 with an additional copy in cis of polh or polhE44G, respectively, under p10 promoter. Our results show that both viruses expressed POLH-Δ3AB1–3, which was detected by western blot in purified rOBs with anti-POLH and anti-3AB1 antibodies. We also found that vPOLHE44G produced larger polyhedra and a significant increase of antigen yield (p < 0.01). Furthermore, the chimeric protein POLH-Δ3AB1–3 was recognized by sera from experimentally infected animals, showing that translational fusion to POLH does not alter the antigenicity of Δ3AB1–3. Finally, the rOBs were successfully used in an ELISA test to differentiate infected from vaccinated animals. Taken together, these results demonstrate the great potential of rOBs to develop diagnostic schemes adaptable to animal infectious diseases.
Instituto de Biotecnología
Fil: Diez, Michay. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina
Fil: Trotta, Myrian Vanesa. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología; Argentina
Fil: Alfonso, Victoria. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Taboga, Oscar Alberto. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Lopez, Maria Gabriela. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Source
3 Biotech 8 : 457. (November 2018)
Subject
Baculovirus
Virus Fiebre Aftosa
Aphthovirus
ELISA
Occlusion Bodies
Foot and Mouth Disease Virus
Polyhedrin
FMDV
DIVA Diagnostic
Cuerpos de Oclusión
Access level
Restricted access
License
Repository
INTA Digital (INTA)
Institution
Instituto Nacional de Tecnología Agropecuaria
OAI Identifier
oai:localhost:20.500.12123/4459