Characterization of the monoclonal antibody against classical swine fever virus glycoprotein E(rns) and its application to an indirect sandwich ELISA

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 prod...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 2011-11, Vol.92 (4), p.815-821
Main Authors: Wu, Ching-Wei, Chien, Maw-Sheng, Liu, Ting-Yu, Lin, Guang-Jan, Lee, Wei-Cheng, Huang, Chienjin
Format: Article
Language:English
Subjects:
Animals
Antibodies, Monoclonal - immunology
Antibodies, Viral - immunology
Antigens, Viral - analysis
Antigens, Viral - immunology
Classical Swine Fever - diagnosis
Classical swine fever virus - immunology
Classical swine fever virus - isolation & purification
Enzyme-Linked Immunosorbent Assay - methods
Epitope Mapping
Glycoproteins - analysis
Glycoproteins - immunology
Pichia - genetics
Recombinant Proteins - analysis
Recombinant Proteins - immunology
Ribonucleases - analysis
Ribonucleases - immunology
Sensitivity and Specificity
Swine
Viral Proteins - analysis
Viral Proteins - immunology
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Summary: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:1432-0614
DOI:10.1007/s00253-011-3602-y