Expression of recombinant West Nile virus prM protein fused to an affinity tag for use as a diagnostic antigen

Setoh, Y. X., Hobson-Peters, J., Prow, N. A., Young, P. R. and Hall, R. A. (2011) Expression of recombinant West Nile virus prM protein fused to an affinity tag for use as a diagnostic antigen. Journal of Virological Methods, 175 1: 20-27. doi:10.1016/j.jviromet.2011.04.010


Author Setoh, Y. X.
Hobson-Peters, J.
Prow, N. A.
Young, P. R.
Hall, R. A.
Title Expression of recombinant West Nile virus prM protein fused to an affinity tag for use as a diagnostic antigen
Journal name Journal of Virological Methods   Check publisher's open access policy
ISSN 0166-0934
1879-0984
Publication date 2011-07
Sub-type Article (original research)
DOI 10.1016/j.jviromet.2011.04.010
Volume 175
Issue 1
Start page 20
End page 27
Total pages 8
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Collection year 2012
Language eng
Formatted abstract
Previous studies have concluded that the Flavivirus prM protein is a suitable viral antigen to distinguish serologically between infections with closely related Flaviviruses (Cardosa et al., 2002). To express the recombinant West Nile virus (WNV) prM antigen fused to a suitable affinity tag for purification, a series of prM-His-tag and prM-V5-tag fusion proteins were generated. Analysis of the prM-His-tag fusion proteins revealed that either prM epitopes were disrupted or the His-tag was not presented properly depending on the location of the His tag and the presence of the prM transmembrane domains in these constructs. This identified domains critical for proper folding of prM, and arrangements that allowed the correct presentation of the His-tag. However, the inclusion of the V5 epitope tag fused to the C terminus of prM allowed formation of the authentic antigenic structure of prM and the proper presentation of the V5 epitope. Capture of tagged recombinant WNVNY99 prM antigen to the solid phase with anti-V5 antibody in ELISA enabled the detection of prM-specific antibodies in WNVNY99-immune horse serum, confirming its potential as a useful diagnostic reagent.
Keyword Flavivirus
prM
West Nile virus
His tag
V5 tag
Diagnostic antigen
Horse serum
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
School of Chemistry and Molecular Biosciences
 
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