title: Characterization of the monoclonal antibody against classical swine fever virus glycoprotein E(rns) and its application to an indirect sandwich ELISA.
abstract: 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.
authors.affiliation: Graduate Institute of Microbiology and Public Health College of Veterinary Medicine, National Chung Hsing University, 250 Kuo Kuang Road, Taichung 40227, Taiwan, Republic of China.
authors.forename: Ching-Wei
authors.initials: CW
authors.lastname: Wu
authors.forename: Maw-Sheng
authors.initials: MS
authors.lastname: Chien
authors.forename: Ting-Yu
authors.initials: TY
authors.lastname: Liu
authors.forename: Guang-Jan
authors.initials: GJ
authors.lastname: Lin
authors.forename: Wei-Cheng
authors.initials: WC
authors.lastname: Lee
authors.forename: Chienjin
authors.initials: C
authors.lastname: Huang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis / Q000276:immunology; D006691:Classical Swine Fever / Q000175:diagnosis*; D006692:Classical Swine Fever Virus / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018604:Epitope Mapping; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D012260:Ribonucleases / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity; D013552:Swine; D014764:Viral Proteins / Q000032:analysis* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D012260:Ribonucleases