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Zika Virus Non-Structural Protein 1 Antigen-Capture Immunoassay

Department of Microbiology and Immunology, School of Medicine, Tulane University, New Orleans, LA 70112, USA
Autoimmune Technologies, Limited Liability Company, New Orleans, LA 70112, USA
Department of Immunology and Microbial Science, Scripps Research, La Jolla, CA 92037, USA
Scripps Translational Science Institute, La Jolla, CA 92037, USA
Department of Integrative Structural and Computational Biology, Scripps Research, La Jolla, CA 92037, USA
MediPath Instituto de Patologia Molecular, Universidad Tecnológica de Santiago (UTESA), Santiago 51000, Dominican Republic
Centro de Investigaciones Biomédicas y Clínicas (CINBIOCLI), Pontificia Universidad Católica Madre y Maestra (PUCMM), Santiago 51034, Dominican Republic
Instituto de Innovacion Biotecnologia e Industria (IIBI), Santo Domingo 10135, Dominican Republic
Ministerio de Salud Publica (MSP), Santo Domingo 10514, Dominican Republic
Zalgen Labs, Limited Liability Company, Germantown, MD 20876, USA
Author to whom correspondence should be addressed.
Current Address: United States Medical Affairs Department, Adult RSV Vaccines Division, The Janssen Pharmaceutical Companies of Johnson and Johnson, Inc., New Brunswick, NJ 08933, USA.
Current Address: Department of Epidemiology of Microbial Diseases, Yale University School of Public Health, New Haven, CT 06510, USA.
Academic Editor: Karla Helbig
Viruses 2021, 13(9), 1771;
Received: 5 August 2021 / Revised: 24 August 2021 / Accepted: 3 September 2021 / Published: 5 September 2021
(This article belongs to the Special Issue Zika Virus: Unanswered Questions)
Infection with Zika virus (ZIKV), a member of the Flavivirus genus of the Flaviviridae family, typically results in mild self-limited illness, but severe neurological disease occurs in a limited subset of patients. In contrast, serious outcomes commonly occur in pregnancy that affect the developing fetus, including microcephaly and other major birth defects. The genetic similarity of ZIKV to other widespread flaviviruses, such as dengue virus (DENV), presents a challenge to the development of specific ZIKV diagnostic assays. Nonstructural protein 1 (NS1) is established for use in immunodiagnostic assays for flaviviruses. To address the cross-reactivity of ZIKV NS1 with proteins from other flaviviruses we used site-directed mutagenesis to modify putative epitopes. Goat polyclonal antibodies to variant ZIKV NS1 were affinity-purified to remove antibodies binding to the closely related NS1 protein of DENV. An antigen-capture ELISA configured with the affinity-purified polyclonal antibody showed a linear dynamic range between approximately 500 and 30 ng/mL, with a limit of detection of between 1.95 and 7.8 ng/mL. NS1 proteins from DENV, yellow fever virus, St. Louis encephalitis virus and West Nile virus showed significantly reduced reactivity in the ZIKV antigen-capture ELISA. Refinement of approaches similar to those employed here could lead to development of ZIKV-specific immunoassays suitable for use in areas where infections with related flaviviruses are common. View Full-Text
Keywords: Zika virus; non-structural protein 1; site-directed mutagenesis; polyclonal antibodies; antigen-capture ELISA Zika virus; non-structural protein 1; site-directed mutagenesis; polyclonal antibodies; antigen-capture ELISA
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MDPI and ACS Style

Beddingfield, B.J.; Hartnett, J.N.; Wilson, R.B.; Kulakosky, P.C.; Andersen, K.G.; Robles-Sikisaka, R.; Grubaugh, N.D.; Aybar, A.; Nunez, M.-Z.; Fermin, C.D.; Garry, R.F. Zika Virus Non-Structural Protein 1 Antigen-Capture Immunoassay. Viruses 2021, 13, 1771.

AMA Style

Beddingfield BJ, Hartnett JN, Wilson RB, Kulakosky PC, Andersen KG, Robles-Sikisaka R, Grubaugh ND, Aybar A, Nunez M-Z, Fermin CD, Garry RF. Zika Virus Non-Structural Protein 1 Antigen-Capture Immunoassay. Viruses. 2021; 13(9):1771.

Chicago/Turabian Style

Beddingfield, Brandon J., Jessica N. Hartnett, Russell B. Wilson, Peter C. Kulakosky, Kristian G. Andersen, Refugio Robles-Sikisaka, Nathan D. Grubaugh, Argelia Aybar, Maria-Zunilla Nunez, Cesar D. Fermin, and Robert F. Garry. 2021. "Zika Virus Non-Structural Protein 1 Antigen-Capture Immunoassay" Viruses 13, no. 9: 1771.

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