title: Characterization of Zaire ebolavirus glycoprotein-specific monoclonal antibodies.
abstract: Zaire ebolavirus (ZEBOV) can be transmitted by human-to-human contact and causes acute haemorrhagic fever with case fatality rates up to 90%. There are no effective therapeutic or prophylactic treatments available. The sole transmembrane glycoprotein (GP) is the key target for developing neutralizing antibodies. In this study, recombinant VSVΔG/ZEBOVGP was used to generate monoclonal antibodies (MAbs) against the ZEBOV GP. A total of 8 MAbs were produced using traditional hybridoma cell fusion technology, and then characterized by ELISA using ZEBOV VLPs, Western blotting, an immunofluorescence assay, and immunoprecipitation. All 8 MAbs worked in IFA and IP, suggesting that they are all conformational MAbs, however six of them recognized linearized epitopes by WB. ELISA results demonstrated that one MAb bound to a secreted GP (sGP 1-295aa); three bind to a part of the mucin domain (333-458aa); three MAbs recognized epitopes on the C-terminal domain of GP1 (296-501aa); and one bound to full length GP (VLPs/GP1,2 ΔTm). Using a mouse model these MAbs were evaluated for their therapeutic capacity during a lethal infection. All 8 MAb improved survival rates by 33%-100% against a high dose lethal challenge with mouse-adapted ZEBOV. This work has important implications for further development of vaccines and immunotherapies for ZEBOV infection.
authors.affiliation: Special Pathogens Program, National Microbiology Laboratory, Public Health Agency of Canada, Winnipeg, Manitoba, Canada. xiangguo.qiu@phac-aspc.gc.ca
authors.forename: Xiangguo
authors.initials: X
authors.lastname: Qiu
authors.forename: Judie B
authors.initials: JB
authors.lastname: Alimonti
authors.forename: P Leno
authors.initials: PL
authors.lastname: Melito
authors.forename: Lisa
authors.initials: L
authors.lastname: Fernando
authors.forename: Ute
authors.initials: U
authors.lastname: Ströher
authors.forename: Steven M
authors.initials: SM
authors.lastname: Jones
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D015153:Blotting, Western; D004306:Dose-Response Relationship, Immunologic; D029043:Ebolavirus / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect; D019142:Hemorrhagic Fever, Ebola / Q000628:therapy; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007116:Immunization, Passive; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000276:immunology*; D047468:Immunoprecipitation; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000276:immunology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D007074:Immunoglobulin G; D014759:Viral Envelope Proteins; C113189:envelope glycoprotein, Ebola virus