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

11 Citations (Scopus)

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

Fingerprint

Ebolavirus
Immunoassay
Antigens
Enzyme-Linked Immunosorbent Assay
Antibodies
Immunoglobulin M
Ebola Hemorrhagic Fever
Marburgvirus
Immunoglobulin G
Sierra Leone
Western Africa
Goats
Primates
Disease Outbreaks
Immunoglobulins
Glycoproteins
Vaccines
Monoclonal Antibodies
Infection
Serum

Keywords

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

ASJC Scopus subject areas

  • Infectious Diseases
  • Immunology and Allergy

Cite this

Boisen, M. L., Oottamasathien, D., Jones, A. B., Millett, M. M., Nelson, D. S., Bornholdt, Z. A., ... 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

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

In: Journal of Infectious Diseases, Vol. 212, 01.10.2015, p. S359-S367.

Research output: Contribution to journalArticle

Boisen, ML, Oottamasathien, D, Jones, AB, Millett, MM, Nelson, DS, Bornholdt, ZA, Fusco, ML, Abelson, DM, Oda, SI, Hartnett, JN, Rowland, MM, Heinrich, ML, Akdag, M, Goba, A, Momoh, M, Fullah, M, Baimba, F, Gbakie, M, Safa, S, Fonnie, R, Kanneh, L, Cross, R, Geisbert, JB, Geisbert, T, Kulakosky, PC, Grant, DS, Shaffer, JG, Schieffelin, JS, Wilson, RB, Saphire, EO, Branco, LM, Garry, RF, Khan, SH & Pitts, KR 2015, 'Development of Prototype Filovirus Recombinant Antigen Immunoassays', Journal of Infectious Diseases, vol. 212, pp. S359-S367. https://doi.org/10.1093/infdis/jiv353
Boisen ML, Oottamasathien D, Jones AB, Millett MM, Nelson DS, Bornholdt ZA et al. Development of Prototype Filovirus Recombinant Antigen Immunoassays. Journal of Infectious Diseases. 2015 Oct 1;212:S359-S367. https://doi.org/10.1093/infdis/jiv353
Boisen, Matt L. ; Oottamasathien, Darin ; Jones, Abigail B. ; Millett, Molly M. ; Nelson, Diana S. ; Bornholdt, Zachary A. ; Fusco, Marnie L. ; Abelson, Dafna M. ; Oda, Shun Ichiro ; Hartnett, Jessica N. ; Rowland, Megan M. ; Heinrich, Megan L. ; Akdag, Marjan ; Goba, Augustine ; Momoh, Mambu ; Fullah, Mohammed ; Baimba, Francis ; Gbakie, Michael ; Safa, Sadiki ; Fonnie, Richard ; Kanneh, Lansana ; Cross, Robert ; Geisbert, Joan B. ; Geisbert, Thomas ; Kulakosky, Peter C. ; Grant, Donald S. ; Shaffer, Jeffery G. ; Schieffelin, John S. ; Wilson, Russell B. ; Saphire, Erica Ollmann ; Branco, Luis M. ; Garry, Robert F. ; Khan, S. Humarr ; Pitts, Kelly R. / Development of Prototype Filovirus Recombinant Antigen Immunoassays. In: Journal of Infectious Diseases. 2015 ; Vol. 212. pp. S359-S367.
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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.",
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T1 - Development of Prototype Filovirus Recombinant Antigen Immunoassays

AU - Boisen, Matt L.

AU - Oottamasathien, Darin

AU - Jones, Abigail B.

AU - Millett, Molly M.

AU - Nelson, Diana S.

AU - Bornholdt, Zachary A.

AU - Fusco, Marnie L.

AU - Abelson, Dafna M.

AU - Oda, Shun Ichiro

AU - Hartnett, Jessica N.

AU - Rowland, Megan M.

AU - Heinrich, Megan L.

AU - Akdag, Marjan

AU - Goba, Augustine

AU - Momoh, Mambu

AU - Fullah, Mohammed

AU - Baimba, Francis

AU - Gbakie, Michael

AU - Safa, Sadiki

AU - Fonnie, Richard

AU - Kanneh, Lansana

AU - Cross, Robert

AU - Geisbert, Joan B.

AU - Geisbert, Thomas

AU - Kulakosky, Peter C.

AU - Grant, Donald S.

AU - Shaffer, Jeffery G.

AU - Schieffelin, John S.

AU - Wilson, Russell B.

AU - Saphire, Erica Ollmann

AU - Branco, Luis M.

AU - Garry, Robert F.

AU - Khan, S. Humarr

AU - Pitts, Kelly R.

PY - 2015/10/1

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N2 - 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.

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KW - Ebola

KW - Ebola virus

KW - ELISA

KW - filovirus

KW - lateral flow immunodiagnostic

KW - point-of-care testing

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