title: Development of Prototype Filovirus Recombinant Antigen Immunoassays.
abstract: BACKGROUND
  Throughout the 2014-2015 Ebola outbreak in West Africa, major gaps were exposed in the availability of validated rapid diagnostic platforms, protective vaccines, and effective therapeutic agents. These gaps potentiated the development of prototype rapid lateral flow immunodiagnostic (LFI) assays that are true point-of-contact platforms, for the detection of active Ebola infections in small blood samples.
  
  
  METHODS
  Recombinant Ebola and Marburg virus matrix VP40 and glycoprotein (GP) antigens were used to derive a panel of monoclonal and polyclonal antibodies. Antibodies were tested using a multivariate approach to identify antibody-antigen combinations suitable for enzyme-linked immunosorbent assay (ELISA) and LFI assay development.
  
  
  RESULTS
  Polyclonal antibodies generated in goats were superior reagents for capture and detection of recombinant VP40 in test sample matrices. These antibodies were optimized for use in antigen-capture ELISA and LFI assay platforms. Prototype immunoglobulin M (IgM)/immunoglobulin G (IgG) ELISAs were similarly developed that specifically detect Ebola virus-specific antibodies in the serum of experimentally infected nonhuman primates and in blood samples obtained from patients with Ebola from Sierra Leone.
  
  
  CONCLUSIONS
  The prototype recombinant Ebola LFI assays developed in these studies have sensitivities that are useful for clinical diagnosis of acute ebolavirus infections. The antigen-capture and IgM/IgG ELISAs provide additional confirmatory assay platforms for detecting VP40 and other ebolavirus-specific immunoglobulins.
authors.affiliation: Corgenix, Broomfield, Colorado Department of Microbiology and Immunology, School of Medicine.
authors.forename: Matt L
authors.initials: ML
authors.lastname: Boisen
authors.affiliation: Corgenix, Broomfield, Colorado.
authors.forename: Darin
authors.initials: D
authors.lastname: Oottamasathien
authors.affiliation: Corgenix, Broomfield, Colorado.
authors.forename: Abigail B
authors.initials: AB
authors.lastname: Jones
authors.affiliation: Corgenix, Broomfield, Colorado.
authors.forename: Molly M
authors.initials: MM
authors.lastname: Millett
authors.affiliation: Corgenix, Broomfield, Colorado.
authors.forename: Diana S
authors.initials: DS
authors.lastname: Nelson
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California.
authors.forename: Zachary A
authors.initials: ZA
authors.lastname: Bornholdt
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California.
authors.forename: Marnie L
authors.initials: ML
authors.lastname: Fusco
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California.
authors.forename: Dafna M
authors.initials: DM
authors.lastname: Abelson
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California.
authors.forename: Shun-Ichiro
authors.initials: S
authors.lastname: Oda
authors.affiliation: Department of Microbiology and Immunology, School of Medicine.
authors.forename: Jessica N
authors.initials: JN
authors.lastname: Hartnett
authors.affiliation: Zalgen Labs, Germantown, Maryland.
authors.forename: Megan M
authors.initials: MM
authors.lastname: Rowland
authors.affiliation: Zalgen Labs, Germantown, Maryland.
authors.forename: Megan L
authors.initials: ML
authors.lastname: Heinrich
authors.affiliation: Zalgen Labs, Germantown, Maryland.
authors.forename: Marjan
authors.initials: M
authors.lastname: Akdag
authors.affiliation: Lassa Fever Program, Kenema Government Hospital.
authors.forename: Augustine
authors.initials: A
authors.lastname: Goba
authors.affiliation: Lassa Fever Program, Kenema Government Hospital Eastern Polytechnic College, Kenema.
authors.forename: Mambu
authors.initials: M
authors.lastname: Momoh
authors.affiliation: Lassa Fever Program, Kenema Government Hospital.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Fullah
authors.affiliation: Lassa Fever Program, Kenema Government Hospital.
authors.forename: Francis
authors.initials: F
authors.lastname: Baimba
authors.affiliation: Section of Infectious Disease, Department of Internal Medicine, School of Medicine, Tulane University.
authors.forename: Michael
authors.initials: M
authors.lastname: Gbakie
authors.affiliation: Lassa Fever Program, Kenema Government Hospital.
authors.forename: Sadiki
authors.initials: S
authors.lastname: Safa
authors.affiliation: Lassa Fever Program, Kenema Government Hospital.
authors.forename: Richard
authors.initials: R
authors.lastname: Fonnie
authors.affiliation: Lassa Fever Program, Kenema Government Hospital.
authors.forename: Lansana
authors.initials: L
authors.lastname: Kanneh
authors.affiliation: Department of Microbiology and Immunology, University of Texas Medical Branch at Galveston.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Cross
authors.affiliation: Department of Microbiology and Immunology, University of Texas Medical Branch at Galveston.
authors.forename: Joan B
authors.initials: JB
authors.lastname: Geisbert
authors.affiliation: Department of Microbiology and Immunology, University of Texas Medical Branch at Galveston.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Geisbert
authors.affiliation: Autoimmune Technologies, New Orleans, Louisiana.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Kulakosky
authors.affiliation: Zalgen Labs, Germantown, Maryland Ministry of Health and Sanitation, Freetown, Sierra Leone.
authors.forename: Donald S
authors.initials: DS
authors.lastname: Grant
authors.affiliation: Department of Biostatistics and Bioinformatics, School of Public Health and Tropical Medicine.
authors.forename: Jeffery G
authors.initials: JG
authors.lastname: Shaffer
authors.affiliation: Section of Infectious Disease, Department of Pediatrics Section of Infectious Disease, Department of Internal Medicine, School of Medicine, Tulane University.
authors.forename: John S
authors.initials: JS
authors.lastname: Schieffelin
authors.affiliation: Autoimmune Technologies, New Orleans, Louisiana.
authors.forename: Russell B
authors.initials: RB
authors.lastname: Wilson
authors.affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California.
authors.forename: Erica Ollmann
authors.initials: EO
authors.lastname: Saphire
authors.affiliation: Zalgen Labs, Germantown, Maryland.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Branco
authors.affiliation: Department of Microbiology and Immunology, School of Medicine Zalgen Labs, Germantown, Maryland.
authors.forename: Robert F
authors.initials: RF
authors.lastname: Garry
authors.affiliation: Lassa Fever Program, Kenema Government Hospital Ministry of Health and Sanitation, Freetown, Sierra Leone.
authors.forename: S Humarr
authors.initials: SH
authors.lastname: Khan
authors.affiliation: Corgenix, Broomfield, Colorado.
authors.forename: Kelly R
authors.initials: KR
authors.lastname: Pitts
mesh_terms: D000354:Africa, Western; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000276:immunology*; D003429:Cross Reactions / Q000276:immunology; D029043:Ebolavirus / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D016563:Filoviridae / Q000276:immunology*; D019142:Hemorrhagic Fever, Ebola / Q000097:blood / Q000276:immunology / Q000821:virology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D007075:Immunoglobulin M / Q000097:blood / Q000276:immunology; D008379:Marburg Virus Disease / Q000097:blood / Q000276:immunology / Q000821:virology; D029024:Marburgvirus / Q000276:immunology; D012807:Sierra Leone
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D007074:Immunoglobulin G; D007075:Immunoglobulin M
keywords: ELISA; Ebola; Ebola virus; filovirus; lateral flow immunodiagnostic; point-of-care testing