Epitope specificity of monoclonal antibodies to the N-protein of porcine reproductive and respiratory syndrome virus determined by ELISA with synthetic peptides.
In an attempt to develop an alternate to ELISAs using recombinant N-proteins as antigen for the sero-diagnosis of porcine reproductive and respiratory syndrome virus (PRRSV) infections of pigs I have measured the binding of nine anti-N-protein mAbs, which had been previously generated by various investigators, to overlapping peptides encompassing amino acids 19-70 of the N-proteins of the North American prototype (VR2332) and the European prototype (Lelystad virus, LV) of PRRSV. I also measured the binding of the mAbs to HerdChek ELISA plates coated with recombinant N-protein. All mAbs bound in an indirect ELISA to some of the peptides whether the mAbs had previously been reported to recognize continuous or discontinuous epitopes, but with different specificity and titer. Three mAbs bound with high titer to different linear epitopes located in amino acid segments 23-33, 31-50 and 43-56 and also with similar high titers to HerdChek plates. mAb SDOW17 bound with high titer to HerdChek plates but poorly to any of the peptides. In contrast, four mAbs bound with broad specificity to peptides containing an epitope(s) in amino acid segment 30-48, but poorly, or not at all, to HerdChek ELISA plates. Thus, this epitope is missing on the antigens of the HerdChek ELISA or is destroyed during immobilization of the antigens on the plate. A mAb to the N-protein of the closely related mouse arterivirus lactate dehydrogenase-elevating virus bound to the same epitope. Abs that bound with broad specificity to an epitope(s) in the 30-50 amino acid segment were also detected by the peptide ELISA in sera of 25 field sera that were sero-positive in the HerdChek ELISA, but also in sera of pigs from two out of three herds tested that were sero-negative by this test.
Plagemann
Peter G W
PG
Department of Microbiology, University of Minnesota, Box 196 UMHC, Minneapolis, MN 55455, USA. plage001@umn.edu
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000097:blood; D001483:Base Sequence; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D000939:Epitopes / Q000276:immunology; D008969:Molecular Sequence Data; D019590:Nucleocapsid Proteins / Q000276:immunology*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis / Q000276:immunology*; D019316:Porcine respiratory and reproductive syndrome virus / Q000276:immunology*; D011933:Reagent Kits, Diagnostic / Q000821:virology; D016415:Sequence Alignment
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D019590:Nucleocapsid Proteins; D011933:Reagent Kits, Diagnostic