IQUIFIB   02644
INSTITUTO DE QUIMICA Y FISICOQUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
Glycoprotein G rabies virus expression in insect cells for diagnostic purposes in vaccinated llamas
Autor/es:
MC CALLUM GREGORIO; MICUCCI MATÍAS; ALFONSO VICTORIA; SMITH IGNACIO; PEREZ OSCAR; FERRARI ALEJANDRO; TARGOVNIK ALEXANDRA; ARREGUI MARIANA; LOPEZ MARÍA GABRIELA; MIRANDA MARÍA VICTORIA
Lugar:
CABA
Reunión:
Conferencia; XXIX Conferencia Internacional de Rabia en las Américas (RITA); 2018
Resumen:
Rabies is one of the principal zoonosis with worldwide distribution. The disease in llamas producesgreat economic losses in the productive system. The prevention against rabies virus (RABV) infectionis done through vaccination with the inactivated virus. The level of protection can be assessed bydetermining the titre of neutralizing serum antibodies induced by RABV glycoprotein (RABV-G) invaccinated individuals, which can be measured using an enzyme-linked immunosorbent assay. Theaim of this work is to optimize a process for the production of the RABV-G ectodomain in Sf9 cell.The recombinant baculovirus was constructed by introducing the sequence corresponding to theectodomain region of RABV-G, under the polyhedrin promoter, fused to the viral signal peptide GP67at its amino terminus and a 6-histidine tag at its carboxyl terminus. Despite the inefficient secretionof the RABV-G, it was possible to recover it from the cell lysate. Insect cells infected at multiplicityof infection of 0.5 at day 3 expressed 7 mg litre-1. The recombinant RABV-G had a molecular massof approximately 50 kDa according to SDS-PAGE analysis. PGnase F and Tunicamycin treatmentconfirmed that the protein is glycosilated. RABV-G was identified by LC-MS and was purified by metalion affinity chromatography directly from the cell extract with a yield of 99% and a purity of 67%.Purified RABV-G was succesfully used to detect specific antibodies in serum samples derived fromrabies-vaccinated llamas. These results indicate that the recombinant RABV-G can be used as anantigen in the development of diagnostic kits