title: Expression and purification of the surface proteins from Andes virus.
abstract: Andes virus is the main causative agent of Hantavirus cardiopulmonary syndrome in South America. There are currently no vaccines or treatments against Andes virus. However, there are several evidences suggesting that antibodies against Andes virus envelope glycoproteins may be enough to confer full protection against Hantavirus cardiopulmonary syndrome. The goal of the present work was to express, purify and characterize the extracellular domains of Andes virus glycoproteins Gn and Gc. We generated two adenoviral vectors encoding the extracellular domains of Andes virus glycoproteins Gn and Gc. Both molecules were expressed by adenoviral transduction in SiHa cells. We found that sGc ectodomain was mainly secreted into the culture medium, whereas sGn was predominantly retained inside the cells. Both molecules were expressed at very low concentrations (below 1 μg/mL). Treatment with the proteasome inhibitor ALLN raised sGc concentration in the cell culture medium, but did not affect expression levels of sGn. Both ectodomains were purified by immobilized metal ion affinity chromatography, and were recognized by sera from persons previously exposed to Andes virus. To our knowledge, this is the first work that addresses the expression and purification of Andes virus glycoproteins Gn and Gc. Our results demonstrate that sGn and sGc maintain epitopes that are exposed on the surface of the viral envelope. However, our work also highlights the need to explore new strategies to achieve high-level expression of these proteins for development of a vaccine candidate against Andes virus.
authors.affiliation: Department of Pharmacology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: Camila E
authors.initials: CE
authors.lastname: Beltrán-Ortiz
authors.affiliation: Department of Pharmacology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: Maria F
authors.initials: MF
authors.lastname: Starck-Mendez
authors.affiliation: Department of Pharmacology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: Yaiza
authors.initials: Y
authors.lastname: Fernández
authors.affiliation: Department of Pharmacology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: Omar
authors.initials: O
authors.lastname: Farnós
authors.affiliation: Department of Physiopathology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: Eddy E
authors.initials: EE
authors.lastname: González
authors.affiliation: Department of Physiopathology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: Coralia I
authors.initials: CI
authors.lastname: Rivas
authors.affiliation: Department of Pharmacology, School of Biological Sciences, University of Concepcion, Chile.
authors.forename: F
authors.initials: F
authors.lastname: Camacho
authors.affiliation: Department of Clinical Biochemistry and Immunology, School of Pharmacia, University of Concepcion, Chile.
authors.forename: Felipe A
authors.initials: FA
authors.lastname: Zuñiga
authors.affiliation: Department of Physiopathology, School of Biological Sciences, University of Concepcion, Chile; Center for Biotechnology and Biomedicine Spa., Chile.
authors.forename: Jorge R
authors.initials: JR
authors.lastname: Toledo
authors.affiliation: Department of Pharmacology, School of Biological Sciences, University of Concepcion, Chile; Center for Biotechnology and Biomedicine Spa., Chile. Electronic address: osanchez@udec.cl.
authors.forename: Oliberto
authors.initials: O
authors.lastname: Sánchez
mesh_terms: D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D006476:Orthohantavirus / Q000235:genetics*; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins
keywords: Andes virus; Gc; Glycoproteins; Gn; Hantavirus