title: Detection of Ebola virus envelope using monoclonal and polyclonal antibodies in ELISA, surface plasmon resonance and a quartz crystal microbalance immunosensor.
abstract: Ebola virus (EBOV) Zaire, Sudan, as well as Ivory Coast are virulent human EBOV species. Both polyclonal and monoclonal antibodies (MAbs) were developed against soluble EBOV envelope glycoprotein (GP) for the study of EBOV envelope diversity and development of diagnostic reagents. Three EBOV Sudan-Gulu GP peptides, from the N-terminus, mid-GP, and C-terminus regions were used to immunize rabbits for the generation of anti-EBOV polyclonal antibodies. Polyclonal antisera raised against the C-terminus peptide could detect both Sudan-Gulu as well as Zaire GPs, while anti-N and mid-region peptide polyclonal sera recognized only EBOV Sudan-Gulu GP. Of the three anti-EBOV GP mouse MAbs produced, MAb 15H10 recognized all human EBOV GP species tested (Zaire, Sudan and Ivory Coast), and as well as reacted with the Reston non-human primate EBOV GPs. In addition, MAb 15H10 bound virion-associated GP of all known EBOV species. MAb 17A3 recognized GPs of both EBOV Sudan-Gulu and Zaire, while MAb 6D11 recognized only EBOV Sudan-Gulu GP. To detect EBOV GP, these antibody reagents were used in ELISA, surface plasmon resonance and in a quartz crystal microbalance immunosensor. Thus, polyclonal and monoclonal antibodies can be used in combination to identify and differentiate both human and non-human primate EBOV GPs.
authors.affiliation: Human Vaccine Institute, Duke University Medical Center, Durham, NC 27710, United States. yu000022@mc.duke.edu
authors.forename: Jae-Sung
authors.initials: JS
authors.lastname: Yu
authors.forename: Hua-Xin
authors.initials: HX
authors.lastname: Liao
authors.forename: Aren E
authors.initials: AE
authors.lastname: Gerdon
authors.forename: Brian
authors.initials: B
authors.lastname: Huffman
authors.forename: Richard M
authors.initials: RM
authors.lastname: Scearce
authors.forename: Mille
authors.initials: M
authors.lastname: McAdams
authors.forename: S Munir
authors.initials: SM
authors.lastname: Alam
authors.forename: Paul M
authors.initials: PM
authors.lastname: Popernack
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Sullivan
authors.forename: David
authors.initials: D
authors.lastname: Wright
authors.forename: David E
authors.initials: DE
authors.lastname: Cliffel
authors.forename: Gary J
authors.initials: GJ
authors.lastname: Nabel
authors.forename: Barton F
authors.initials: BF
authors.lastname: Haynes
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D003429:Cross Reactions; D029043:Ebolavirus / Q000145:classification / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D020349:Surface Plasmon Resonance / Q000379:methods*; D014759:Viral Envelope Proteins / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014759:Viral Envelope Proteins; C113189:envelope glycoprotein, Ebola virus