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Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development
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dc.contributor.author | Lo, Yi-Ting | |
dc.contributor.author | Tulloch, Fiona | |
dc.contributor.author | Wu, Hsing-Chieh | |
dc.contributor.author | Luke, Garry A. | |
dc.contributor.author | Ryan, Martin D. | |
dc.contributor.author | Chu, Chun-Yen | |
dc.date.accessioned | 2022-03-03T00:39:47Z | |
dc.date.available | 2022-03-03T00:39:47Z | |
dc.date.issued | 2021-09 | |
dc.identifier | 273006510 | |
dc.identifier | d262e054-9116-4c0c-aa31-d9b6e1fbbf81 | |
dc.identifier | 000624628000001 | |
dc.identifier | 85101896800 | |
dc.identifier.citation | Lo , Y-T , Tulloch , F , Wu , H-C , Luke , G A , Ryan , M D & Chu , C-Y 2021 , ' Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development ' , Journal of Applied Microbiology , vol. 131 , no. 3 , pp. 1123-1135 . https://doi.org/10.1111/jam.15044 | en |
dc.identifier.issn | 1364-5072 | |
dc.identifier.other | RIS: urn:D5411F54A7C0D6D6D83A00B5E051DD60 | |
dc.identifier.other | ORCID: /0000-0002-0012-0614/work/90111661 | |
dc.identifier.other | ORCID: /0000-0003-0859-2867/work/90112126 | |
dc.identifier.uri | https://hdl.handle.net/10023/24987 | |
dc.description | This study was support by grants from the Taiwan Ministry of Science and Technology (MOST-106-2911-I-020-501; MOST-107-2313-B-020-011-MY3) and the UK Biotechnology and Biological Sciences Research Council (BB/P025080/1). | en |
dc.description.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. | |
dc.format.extent | 12 | |
dc.format.extent | 31146932 | |
dc.language.iso | eng | |
dc.relation.ispartof | Journal of Applied Microbiology | en |
dc.subject | Bovine ephemeral fever virus | en |
dc.subject | G glycoprotein | en |
dc.subject | Vaccine | en |
dc.subject | Infectious diseases | en |
dc.subject | QR180 Immunology | en |
dc.subject | NDAS | en |
dc.subject | SDG 3 - Good Health and Well-being | en |
dc.subject.lcc | QR180 | en |
dc.title | Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development | en |
dc.type | Journal article | en |
dc.contributor.sponsor | BBSRC | en |
dc.contributor.institution | University of St Andrews. School of Biology | en |
dc.contributor.institution | University of St Andrews. Biomedical Sciences Research Complex | en |
dc.identifier.doi | 10.1111/jam.15044 | |
dc.description.status | Peer reviewed | en |
dc.date.embargoedUntil | 2022-03-03 | |
dc.identifier.grantnumber | BB/P025080/1 | en |
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