Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development

Yi-Ting Lo, Fiona Tulloch, Hsing-Chieh Wu, Garry A. Luke, Martin D. Ryan, Chun-Yen Chu

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Abstract

Aims Vaccines for bovine ephemeral fever virus (BEFV) are available but are difficult to produce, expensive, or suffer from genetic instability. Therefore, we designed constructs encoding C-terminally truncated forms (transmembrane anchoring region deleted) of glycoproteins G and GNS such that they were secreted from the cell into the media to achieve high-level antigen expression, correct glycosylation pattern, and enable further simple purification with the V5 epitope tag.
Methods and Results In this study, synthetic biology was employed to create membrane-bound and secreted forms of G and GNS glycoprotein. Mammalian cell culture was employed as an antigen expression platform, and the secreted forms of G and GNS protein were easily purified from media by using a highly effective, single-step method. The V5 epitope tag was genetically fused to the C-termini of the proteins, enabling detection of the antigen through immunoblotting and immunomicroscopy. Our data demonstrated that the C-terminally truncated form of the G glycoprotein was efficiently secreted from cells into the cell media. Moreover, the immunogenicity was confirmed in mice test.
Conclusions The immuno-dot blots showed that the truncated G glycoprotein was present in the total cell extract, and was clearly secreted into the media, consistent with the western blotting data and live-cell images. Our strategy presented the expression of secreted, epitope-tagged, forms of the BEFV glycoproteins such that appropriately glycosylated forms of BEFV G protein was secreted from the BHK-21 cells. This indicates that high-level expression of secreted G glycoprotein is a feasible strategy for large-scale production of vaccines and improving vaccine efficacy. Significance and Impact of the Study The antigen expression strategy designed in this study can produce high-quality recombinant protein and reduce the amount of antigen used in the vaccine.
Original languageEnglish
Pages (from-to)1123-1135
Number of pages12
JournalJournal of Applied Microbiology
Volume131
Issue number3
Early online date3 Mar 2021
DOIs
Publication statusPublished - Sept 2021

Keywords

  • Bovine ephemeral fever virus
  • G glycoprotein
  • Vaccine
  • Infectious diseases

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