Development of an improved Epstein-Barr virus (EBV) neutralizing antibody assay to facilitate development of a prophylactic gp350-subunit EBV vaccine

Hui Liu; Lorraine Gemmell; Rui Lin; Fengrong Zuo; Henry H. Balfour; Jennifer C. Woo; Gregory M. Hayes

doi:10.4084/MJHID.2020.016

Development of an improved Epstein-Barr virus (EBV) neutralizing antibody assay to facilitate development of a prophylactic gp350-subunit EBV vaccine

Hui Liu, Lorraine Gemmell, Rui Lin, Fengrong Zuo, Henry H. Balfour, Jennifer C. Woo, Gregory M. Hayes

Laboratory Medicine and Pathology

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

4 Scopus citations

Abstract

No licensed vaccine is available for prevention of EBV-associated diseases, and robust, high-throughput bioanalytical assays are needed to evaluate immunogenicity of gp350 subunit-based candidate EBV vaccines. Here we have developed an improved EBV-GFP based neutralization assay for such a vaccine's pre-clinical and clinical validation to measure EBV specific neutralizing antibodies in human donors. The supplementation of guinea pig complement of our previously published high-throughput EBV-GFP fluorescent focus (FFA)based neutralization assay allowed the detection of complement-dependent neutralizing antibodies using a panel of heat-inactivated healthy human sera. Anti-gp350 antibody titers, which were evaluated using a previously optimized anti-gp350 IgG ELISA assay, were moderately correlated to the FFA-based neutralization titers. Overall, this improved high-throughput neutralization assay is capable of characterizing the serologic neutralizing antibody response to natural EBV infection, with applications in evaluating EBV antibody status in epidemiologic studies and immunogenicity of candidate gp350-subunit EBV vaccines in clinical studies.

Original language	English (US)
Article number	e2020016
Journal	Mediterranean Journal of Hematology and Infectious Diseases
Volume	12
Issue number	1
DOIs	https://doi.org/10.4084/MJHID.2020.016
State	Published - 2020

Bibliographical note

Funding Information:
We thank Dr. Lindsey Hutt-Fletcher (Louisiana State University) for providing Akata-BX1-g cell line and SVK-CR2 cell line. We also thank Hong Jin (MedImmune/AstraZeneca) for editing the manuscript.

Publisher Copyright:
© 2020 Universita Cattolica del Sacro Cuore. All rights reserved.

Keywords

Anti-gp350 IgG ELISA
Epstein-Barr Virus
Gp350
Neutralization assay
Vaccine

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title = "Development of an improved Epstein-Barr virus (EBV) neutralizing antibody assay to facilitate development of a prophylactic gp350-subunit EBV vaccine",

abstract = "No licensed vaccine is available for prevention of EBV-associated diseases, and robust, high-throughput bioanalytical assays are needed to evaluate immunogenicity of gp350 subunit-based candidate EBV vaccines. Here we have developed an improved EBV-GFP based neutralization assay for such a vaccine's pre-clinical and clinical validation to measure EBV specific neutralizing antibodies in human donors. The supplementation of guinea pig complement of our previously published high-throughput EBV-GFP fluorescent focus (FFA)based neutralization assay allowed the detection of complement-dependent neutralizing antibodies using a panel of heat-inactivated healthy human sera. Anti-gp350 antibody titers, which were evaluated using a previously optimized anti-gp350 IgG ELISA assay, were moderately correlated to the FFA-based neutralization titers. Overall, this improved high-throughput neutralization assay is capable of characterizing the serologic neutralizing antibody response to natural EBV infection, with applications in evaluating EBV antibody status in epidemiologic studies and immunogenicity of candidate gp350-subunit EBV vaccines in clinical studies.",

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AU - Zuo, Fengrong

AU - Balfour, Henry H.

AU - Woo, Jennifer C.

AU - Hayes, Gregory M.

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N2 - No licensed vaccine is available for prevention of EBV-associated diseases, and robust, high-throughput bioanalytical assays are needed to evaluate immunogenicity of gp350 subunit-based candidate EBV vaccines. Here we have developed an improved EBV-GFP based neutralization assay for such a vaccine's pre-clinical and clinical validation to measure EBV specific neutralizing antibodies in human donors. The supplementation of guinea pig complement of our previously published high-throughput EBV-GFP fluorescent focus (FFA)based neutralization assay allowed the detection of complement-dependent neutralizing antibodies using a panel of heat-inactivated healthy human sera. Anti-gp350 antibody titers, which were evaluated using a previously optimized anti-gp350 IgG ELISA assay, were moderately correlated to the FFA-based neutralization titers. Overall, this improved high-throughput neutralization assay is capable of characterizing the serologic neutralizing antibody response to natural EBV infection, with applications in evaluating EBV antibody status in epidemiologic studies and immunogenicity of candidate gp350-subunit EBV vaccines in clinical studies.

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Development of an improved Epstein-Barr virus (EBV) neutralizing antibody assay to facilitate development of a prophylactic gp350-subunit EBV vaccine

Abstract

Bibliographical note

Keywords

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