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Development of a robust, higher throughput green fluorescent protein (GFP)-based Epstein-Barr Virus (EBV) micro-neutralization assay.
Lin, Rui; Heeke, Darren; Liu, Hui; Rao, Eileen; Marshall, Jason D; Chio, Vera; Cataniag, Floro; Yu, Li; Zuo, Fengrong; McCarthy, Michael P.
Afiliação
  • Lin R; Applied Immunology and Microbiology Group, MedImmune, Mountain View, CA, USA.
  • Heeke D; Applied Immunology and Microbiology Group, MedImmune, Mountain View, CA, USA.
  • Liu H; Applied Immunology and Microbiology Group, MedImmune, Mountain View, CA, USA.
  • Rao E; Translational Biology Group, MedImmune, Mountain View, CA, USA.
  • Marshall JD; Vaccine Platform Group, MedImmune, Gaithersburg, MD, USA.
  • Chio V; Applied Immunology and Microbiology Group, MedImmune, Mountain View, CA, USA.
  • Cataniag F; Vaccine Platform Group, MedImmune, Gaithersburg, MD, USA.
  • Yu L; Statistical Sciences, MedImmune, Gaithersburg, MD, USA.
  • Zuo F; Applied Immunology and Microbiology Group, MedImmune, Mountain View, CA, USA.
  • McCarthy MP; Vaccine Platform Group, MedImmune, Gaithersburg, MD, USA. Electronic address: mikecee2@verizon.net.
J Virol Methods ; 247: 15-21, 2017 09.
Article em En | MEDLINE | ID: mdl-28457783
The goal of most prophylactic vaccines is to elicit robust and effective neutralizing antibodies against the human pathogen target. The titer of neutralizing antibodies to Epstein-Barr Virus (EBV) is a useful biomarker for evaluating EBV vaccines. Here, the development and optimization of a 96-well micro-neutralization fluorescent imaging assay (FIA) using an EBV virus-encoding green fluorescent protein (GFP) to infect adherent EBV recipient cells is reported. The conditions were optimized for generating reproducible EBV-GFP virus, for maintaining viral infectivity for months, and for efficient viral infection of recipient cell culture. The utility of the EBV-GFP FIA neutralization assay was demonstrated in a mouse study of an investigational adjuvanted EBV gp350 subunit vaccine. This assay confirmed the generation of high titers of anti-EBV-neutralizing antibodies which correlated well with the established Raji cell-based flow cytometry-based EBV neutralization assay, as well as with anti-gp350 IgG titers. In naturally infected EBV+ human serum samples, a good correlation between anti-gp350 IgG ELISA titer and EBV-GFP FIA neutralization antibody titer was also observed. Taken together, these results demonstrate the establishment of a scalable high throughput EBV-GFP FIA micro-neutralization assay suitable to measure humoral EBV vaccine response in a large-scale human trial.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Testes de Neutralização / Herpesvirus Humano 4 / Proteínas de Fluorescência Verde / Ensaios de Triagem em Larga Escala / Anticorpos Antivirais Limite: Animals Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Testes de Neutralização / Herpesvirus Humano 4 / Proteínas de Fluorescência Verde / Ensaios de Triagem em Larga Escala / Anticorpos Antivirais Limite: Animals Idioma: En Ano de publicação: 2017 Tipo de documento: Article