Development of Prototype Filovirus Recombinant Antigen Immunoassays

Matt L. Boisen, Darin Oottamasathien, Abigail B. Jones, Molly M. Millett, Diana S. Nelson, Zachary A. Bornholdt, Marnie L. Fusco, Dafna M. Abelson, Shun Ichiro Oda, Jessica N. Hartnett, Megan M. Rowland, Megan L. Heinrich, Marjan Akdag, Augustine Goba, Mambu Momoh, Mohammed Fullah, Francis Baimba, Michael Gbakie, Sadiki Safa, Richard FonnieLansana Kanneh, Robert Cross, Joan B. Geisbert, Thomas Geisbert, Peter C. Kulakosky, Donald S. Grant, Jeffery G. Shaffer, John S. Schieffelin, Russell B. Wilson, Erica Ollmann Saphire, Luis M. Branco, Robert F. Garry, S. Humarr Khan, Kelly R. Pitts

Research output: Contribution to journalArticle

12 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)S359-S367
JournalJournal of Infectious Diseases
Volume212
DOIs
StatePublished - Oct 1 2015

Keywords

  • Ebola
  • Ebola virus
  • ELISA
  • filovirus
  • lateral flow immunodiagnostic
  • point-of-care testing

ASJC Scopus subject areas

  • Infectious Diseases
  • Immunology and Allergy

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    Boisen, M. L., Oottamasathien, D., Jones, A. B., Millett, M. M., Nelson, D. S., Bornholdt, Z. A., Fusco, M. L., Abelson, D. M., Oda, S. I., Hartnett, J. N., Rowland, M. M., Heinrich, M. L., Akdag, M., Goba, A., Momoh, M., Fullah, M., Baimba, F., Gbakie, M., Safa, S., ... Pitts, K. R. (2015). Development of Prototype Filovirus Recombinant Antigen Immunoassays. Journal of Infectious Diseases, 212, S359-S367. https://doi.org/10.1093/infdis/jiv353