Skip to content

Research at St Andrews

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

Research output: Contribution to journalArticlepeer-review

DOI

Standard

Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development. / Lo, Yi-Ting; Tulloch, Fiona; Wu, Hsing-Chieh; Luke, Garry A.; Ryan, Martin D.; Chu, Chun-Yen.

In: Journal of Applied Microbiology, Vol. Early View, 03.03.2021.

Research output: Contribution to journalArticlepeer-review

Harvard

Lo, Y-T, Tulloch, F, Wu, H-C, Luke, GA, Ryan, MD & 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. Early View. https://doi.org/10.1111/jam.15044

APA

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, Early View. https://doi.org/10.1111/jam.15044

Vancouver

Lo Y-T, Tulloch F, Wu H-C, Luke GA, Ryan MD, Chu C-Y. Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development. Journal of Applied Microbiology. 2021 Mar 3;Early View. https://doi.org/10.1111/jam.15044

Author

Lo, Yi-Ting ; Tulloch, Fiona ; Wu, Hsing-Chieh ; Luke, Garry A. ; Ryan, Martin D. ; Chu, Chun-Yen. / Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development. In: Journal of Applied Microbiology. 2021 ; Vol. Early View.

Bibtex - Download

@article{d262e05491164c0caa31d9b6e1fbbf81,
title = "Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development",
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.",
keywords = "Bovine ephemeral fever virus, G glycoprotein, Vaccine, Infectious diseases",
author = "Yi-Ting Lo and Fiona Tulloch and Hsing-Chieh Wu and Luke, {Garry A.} and Ryan, {Martin D.} and Chun-Yen Chu",
note = "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).",
year = "2021",
month = mar,
day = "3",
doi = "10.1111/jam.15044",
language = "English",
volume = "Early View",
journal = "Journal of Applied Microbiology",
issn = "1364-5072",
publisher = "Wiley-Blackwell",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development

AU - Lo, Yi-Ting

AU - Tulloch, Fiona

AU - Wu, Hsing-Chieh

AU - Luke, Garry A.

AU - Ryan, Martin D.

AU - Chu, Chun-Yen

N1 - 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).

PY - 2021/3/3

Y1 - 2021/3/3

N2 - 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.

AB - 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.

KW - Bovine ephemeral fever virus

KW - G glycoprotein

KW - Vaccine

KW - Infectious diseases

U2 - 10.1111/jam.15044

DO - 10.1111/jam.15044

M3 - Article

VL - Early View

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

ER -

Related by author

  1. Translation of Viral Proteins

    Ryan, M. D. & Luke, G. A., 2021, Encyclopedia of Virology: The Virus as a Concept - Fundamentals of Virology. Bamford, D. & Zuckerman, M. (eds.). 4 ed. New York: ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, Vol. 1. p. 444-459 15 p.

    Research output: Chapter in Book/Report/Conference proceedingChapter

  2. A transgenic line that reports CSF1R protein expression provides a definitive marker for the mouse mononuclear phagocyte system

    Grabert, K., Sehgal, A., Irvine, K. M., Wollscheid-Lengeling, E., Ozdemir, D. D., Stables, J., Luke, G. A., Ryan, M. D., Adamson, A., Humphreys, N. E., Sandrock, C. J., Rojo, R., Verkasalo, V. A., Mueller, W., Hohenstein, P., Pettit, A. R., Pridans, C. & Hume, D. A., 1 Dec 2020, In: The Journal of Immunology. 205, 11

    Research output: Contribution to journalArticlepeer-review

  3. Therapeutic applications of the 'NPGP' family of viral 2As

    Luke, G. A. & Ryan, M. D., Nov 2018, In: Reviews in Medical Virology. 28, 6, 12 p., e2001.

    Research output: Contribution to journalReview articlepeer-review

  4. Problems in FMD eradication: a way forward?

    Tulloch, F., Luke, G. A. & Ryan, M. D., 5 Jul 2018, In: Journal of Veterinary Medicine and Research. 5, 5, 6 p., 1140.

    Research output: Contribution to journalReview articlepeer-review

Related by journal

  1. A metaproteomic approach gives functional insights into anaerobic digestion

    Abram, F., Enright, A. -M., O'Reilly, J., Botting, C. H., Collins, G. & O'Flaherty, V., Jun 2011, In: Journal of Applied Microbiology. 110, 6, p. 1550-1560 11 p.

    Research output: Contribution to journalArticlepeer-review

  2. Molecular methods for Mycobacterium tuberculosis strain typing: A users guide

    Kanduma, E., McHugh, T. D. & Gillespie, S. H., 20 May 2003, In: Journal of Applied Microbiology. 94, 5, p. 781-791 11 p.

    Research output: Contribution to journalReview articlepeer-review

  3. A study of the minimum inhibitory concentration and mode of action of oregano essential oil, thymol and carvacrol

    Lambert, RJW., Skandamis, PN., Coote, P. J. & Nychas, G. J. E., Sep 2001, In: Journal of Applied Microbiology. 91, p. 453 - 462 10 p.

    Research output: Contribution to journalArticlepeer-review

ID: 273006510

Top