Development of a quantitative enzyme linked immunosorbent assay for monitoring the Enterovirus 71 vaccine manufacturing process

Chia Chyi Liu, Hsuen Wen Chang, Grace Yang, Jen Ron Chiang, Yen Hung Chow, I. Hsi Sai, Jui Yuan Chang, Sue Chen Lin, Charles Sia, Chia Hsin Hsiao, Ai Hsiang Chou, Pele Chong

Research output: Contribution to journalArticlepeer-review

18 Citations (Scopus)

Abstract

Enterovirus 71 (EV71), the etiologic agent causes outbreaks with significant mortality in young children in Asia and currently there is no vaccine available. In this study, we report a quantitative enzyme linked immunosorbent assay (Q-ELISA) to determine the concentration of the EV71 VP2 antigen. EV71 virus-like particles (VLPs) were produced in the baculovirus expression system and used as the EV71 antigen reference standard. Antisera from both EV71-immunized chickens and rabbits were very efficient and useful as capture antibodies to bind various forms of EV71 antigens, whereas a commercial VP2-specific virus neutralizing monoclonal antibody MAB979 was found to be suitable for quantifying the amount of VP2 antigen. This Q-ELISA was used successfully to determine VP2 content at each stage of EV71 vaccine manufacturing process, particularly during the upstream harvest, downstream purification and viral inactivation steps. The amount of VP2 antigen and the magnitude of neutralizing titers were found to be dose-dependent in mice immunized with vaccine candidates. These results indicate that Q-ELISA could provide off-line timely quantitative measurements of VP2 antigen throughout the production cycle to evaluate critical attributes and conditions that may affect virus yields in culture media, the quality of purification methods, the stability and potency of final vaccine formulations.

Original languageEnglish
Pages (from-to)60-68
Number of pages9
JournalJournal of Virological Methods
Volume176
Issue number1-2
DOIs
Publication statusPublished - Sep 1 2011
Externally publishedYes

Keywords

  • ELISA
  • Enterovirus 71
  • Neutralization epitope
  • Process analytical technology
  • Viral antigen

ASJC Scopus subject areas

  • Virology

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