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 Fonnie; Lansana Kanneh; Robert W. Cross; Joan B. Geisbert; Thomas W. 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

doi:10.1093/infdis/jiv353

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 W. Cross, Joan B. Geisbert, Thomas W. 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

Microbiology And Immunology

Research output: Contribution to journal › Article › peer-review

30 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 language	English (US)
Pages (from-to)	S359-S367
Journal	Journal of Infectious Diseases
Volume	212
DOIs	https://doi.org/10.1093/infdis/jiv353
State	Published - Oct 1 2015

Keywords

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

ASJC Scopus subject areas

General Medicine

Access to Document

10.1093/infdis/jiv353

Cite this

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

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, RW, Geisbert, JB, Geisbert, TW, 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

@article{84b3be3bbf9e456c983a9b2d7a790b6b,

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.",

keywords = "ELISA, Ebola, Ebola virus, filovirus, lateral flow immunodiagnostic, point-of-care testing",

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

note = "Publisher Copyright: {\textcopyright} 2015 The Author 2015.",

year = "2015",

month = oct,

day = "1",

doi = "10.1093/infdis/jiv353",

language = "English (US)",

volume = "212",

pages = "S359--S367",

journal = "Journal of Infectious Diseases",

issn = "0022-1899",

publisher = "Oxford University Press",

}

TY - JOUR

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 W.

AU - Geisbert, Joan B.

AU - Geisbert, Thomas W.

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

Y1 - 2015/10/1

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.

AB - 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.

KW - ELISA

KW - Ebola

KW - Ebola virus

KW - filovirus

KW - lateral flow immunodiagnostic

KW - point-of-care testing

UR - http://www.scopus.com/inward/record.url?scp=84943231423&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84943231423&partnerID=8YFLogxK

U2 - 10.1093/infdis/jiv353

DO - 10.1093/infdis/jiv353

M3 - Article

C2 - 26232440

AN - SCOPUS:84943231423

SN - 0022-1899

VL - 212

SP - S359-S367

JO - Journal of Infectious Diseases

JF - Journal of Infectious Diseases

ER -

Development of Prototype Filovirus Recombinant Antigen Immunoassays

Abstract

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this