title: Production, purification and immunogenicity of recombinant Ebola virus proteins - A comparison of Freund's adjuvant and adjuvant system 03.
abstract: There is an urgent need for Ebola virus (EBOV) proteins, EBOV-specific antibodies and recombinant antigens to be used in diagnostics and as potential vaccine candidates. Our objective was to produce and purify recombinant proteins for immunological assays and for the production of polyclonal EBOV specific antibodies. In addition, a limited comparison of the adjuvant effects of Freund's complete adjuvant (FCA) and adjuvant system 03 (AS03) was carried out. Recombinant EBOV GST-VP24, -VP30, -VP35, -VP40 and -NP were produced in E. coli and purified with affinity chromatography followed by preparative gel electrophoresis. Recombinant EBOV GP-His was produced in Sf9 insect cells and purified by preparative gel electrophoresis. To compare the adjuvant effect of FCA and AS03, 12 rabbits were immunized four times with one of the six recombinant EBOV proteins using FCA or AS03. In addition, three guinea pigs were immunized with EBOV VP24 using FCA. With the exception of sera from two rabbits immunized with GST-VP24, the antisera against all other EBOV proteins showed very high and specific antibody responses after three to four immunizations. The adjuvant effect of AS03 was comparable to that of FCA. The produced antibodies recognized the corresponding EBOV proteins in wild type EBOV-infected cells.
authors.affiliation: Department of Virology, University of Turku, Turku, Finland; Viral Infections Unit, National Institute for Health and Welfare, Helsinki, Finland. Electronic address: krister.melen@thl.fi.
authors.forename: Krister
authors.initials: K
authors.lastname: Melén
authors.affiliation: Department of Virology, University of Turku, Turku, Finland.
authors.forename: Laura
authors.initials: L
authors.lastname: Kakkola
authors.affiliation: Department of Virology, University of Turku, Turku, Finland.
authors.forename: Felix
authors.initials: F
authors.lastname: He
authors.affiliation: Department of Biotechnology and Molecular Medicine, University of Eastern Finland, Kuopio, Finland.
authors.forename: Kari
authors.initials: K
authors.lastname: Airenne
authors.affiliation: Departments of Virology and Veterinary Biosciences, University of Helsinki, Helsinki, Finland; Department of Virology and Immunology, Helsinki University Hospital, Helsinki, Finland.
authors.forename: Olli
authors.initials: O
authors.lastname: Vapalahti
authors.affiliation: Public Health Agency of Sweden, Solna, Sweden.
authors.forename: Helen
authors.initials: H
authors.lastname: Karlberg
authors.affiliation: Public Health Agency of Sweden, Solna, Sweden; LABMED, Clinical Microbiology, Karolinska Institutet and karolinska university hospital, Stockholm, Sweden; National Veterinary Institute, Uppsala, Sweden.
authors.forename: Ali
authors.initials: A
authors.lastname: Mirazimi
authors.affiliation: Department of Virology, University of Turku, Turku, Finland; Viral Infections Unit, National Institute for Health and Welfare, Helsinki, Finland. Electronic address: ilkka.julkunen@utu.fi.
authors.forename: Ilkka
authors.initials: I
authors.lastname: Julkunen
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000097:blood / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics; D004338:Drug Combinations; D029043:Ebolavirus / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D005620:Freund's Adjuvant / Q000600:supply & distribution; D006168:Guinea Pigs; D011136:Polysorbates; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D013185:Squalene; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D024502:alpha-Tocopherol
chemical_list: D000276:Adjuvants, Immunologic; D000914:Antibodies, Viral; D004338:Drug Combinations; D011136:Polysorbates; D011994:Recombinant Proteins; D014764:Viral Proteins; D013185:Squalene; D005620:Freund's Adjuvant; C550253:AS03 adjuvant; D024502:alpha-Tocopherol
keywords: Adjuvant; Antibody; Baculovirus expression; Ebola virus; Protein purification