Identification of an immunodominant epitope in the C terminus of glycoprotein 5 of porcine reproductive and respiratory syndrome virus.
Glycoprotein 5 (GP(5)) is the major glycoprotein of porcine reproductive and respiratory syndrome virus (PRRSV). Expression of GP(5) has been improved by removing the transmembrane regions. Vectors were constructed encoding complete GP(5) plus three mutants: GP(5) Ns (residues 28--201), GP(5)[30--67] (residues 30--67) and GP(5)[30--201] (residues 30--67/130--201). The three deletion mutants were expressed at levels 20--30 times higher than complete GP(5). GP(5)[30--201] was well recognized in ELISA or immunoblotting by a collection of pig sera. All the fragments were tested for the generation of MAbs, but only the polyhistidine-tagged fragment GP(5)[30--201]H elicited an antibody response sufficient to produce MABS: The two MAbs were positive for PRRSV in ELISA and immunoblotting, but negative for virus neutralization. MAb 4BE12 reacted with residues 130--170 and MAb 3AH9 recognized residues 170--201. This region was recognized strongly in immunoblotting by a collection of infected-pig sera. These results indicate diagnostic potential for this epitope.
Rodriguez
María Jose
MJ
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
Sarraseca
Javier
J
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
Fominaya
Jesús
J
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
Cortés
Elena
E
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
Sanz
Antonio
A
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
Casal
J Ignacio
JI
INGENASA, Hermanos Garcia Noblejas 41, 28037 Madrid, Spain1.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D016296:Mutagenesis; D009500:Neutralization Tests; D019318:Porcine Reproductive and Respiratory Syndrome / Q000097:blood / Q000276:immunology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000276:immunology*; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D018985:Epitopes, B-Lymphocyte; D016056:Immunodominant Epitopes; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; C114789:glycoprotein 5, PRRSV