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標題: Characterization of the monoclonal antibody against classical swine fever virus glycoprotein E-rns and its application to an indirect sandwich ELISA
作者: Wu, C.W.
Chien, M.S.
Liu, T.Y.
Lin, G.J.
Lee, W.C.
Huang, C.J.
關鍵字: Classical swine fever virus;RNase activity;Monoclonal antibody;Sandwich ELISA;rnase activity;pestivirus;vaccine;identification;ribonuclease;expression;immunity
Project: Applied Microbiology and Biotechnology
期刊/報告no:: Applied Microbiology and Biotechnology, Volume 92, Issue 4, Page(s) 815-821.
Classical swine fever virus (CSFV) E-rns is an envelope glycoprotein possessing RNase activity. The E-rns-based enzyme-linked immunosorbent assay (ELISA) has been considered a discriminating diagnostic test for differentiating infected from vaccinated animals. The purpose of this study was to produce a specific monoclonal antibody (MAb) to E-rns for further developing an indirect sandwich ELISA. The MAb CW813 was shown to specifically recognize both the monomer and dimer forms of Pichia pastoris yeast-expressed E-rns (yE(rns)). The antigenic site recognized by MAb CW813 was mapped to the region of amino acid residues 101-160 of E-rns where it was neither a neutralizing epitope nor essential to RNase activity. Furthermore, MAb CW813 was utilized as a capture antibody to develop a yE(rns)-based indirect sandwich ELISA for detecting swine antibody to E-rns. The assay demonstrated a high sensitivity and specificity that may provide an alternative method for developing a diagnostic kit with easy manipulation and low cost.
ISSN: 0175-7598
DOI: 10.1007/s00253-011-3602-y
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