Abstract
Classical swine fever virus (CSFV) Erns is an envelope glycoprotein possessing RNase activity. The Erns-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 Erns 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 Erns (yErns). The antigenic site recognized by MAb CW813 was mapped to the region of amino acid residues 101–160 of Erns where it was neither a neutralizing epitope nor essential to RNase activity. Furthermore, MAb CW813 was utilized as a capture antibody to develop a yErns-based indirect sandwich ELISA for detecting swine antibody to Erns. 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.
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Acknowledgments
This work was supported in part by the grant NSC97-2323-B-005-007-MY3 from the National Science Council and the grant 99AS-9.2.5-BQ-B1(5) from the Bureau of Animal and Plant Health Inspection and Quarantine, Council of Agriculture, Taiwan, Republic of China.
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Wu, CW., Chien, MS., Liu, TY. et al. Characterization of the monoclonal antibody against classical swine fever virus glycoprotein Erns and its application to an indirect sandwich ELISA. Appl Microbiol Biotechnol 92, 815–821 (2011). https://doi.org/10.1007/s00253-011-3602-y
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DOI: https://doi.org/10.1007/s00253-011-3602-y