Monoclonal antibodies against conformationally dependent epitopes on porcine reproductive and respiratory syndrome virus.
Monoclonal antibodies (MAbs) against porcine reproductive and respiratory syndrome virus (PRRSV) were prepared and characterized. Four MAbs were developed from the mice immunized with the recombinant GP4 protein expressed in insect cells, and six MAbs were derived from the immunization with recombinant GP5 protein. All of the MAbs showed strong perinuclear fluorescence in PRRSV VR2385 infected cells by immunofluorescence staining. Among the MAbs to GP5 protein, one showed strong reactivity in ELISA and recognized a 26 kDa band of PRRSV in a western blot assay, while another showed neutralizing activity against the VR2385 isolate. Out of the four MAbs to GP4 protein, one showed mild reactivity in ELISA with detergent extracted antigen, but had no reactivity in a western-blot assay. The failure of MAb binding to detergent extracted antigen in ELISA or in western-blot analysis indicated that the MAbs were against conformationally dependent epitopes. Reactivity patterns of the MAbs with PRRSV field isolates tested by fixed-cell ELISA showed that there are antigenic variations in PRRSV GP4 and GP5 proteins. Development of these MAbs will benefit further studies on PRRSV structural proteins as well as in understanding their roles in PRRSV pathogenesis.
Zhang
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Y
Veterinary Medical Research Institute, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
Sharma
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RD
Paul
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PS
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology*; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry; D019084:Fluorescent Antibody Technique, Indirect; D051379:Mice; D016366:Open Reading Frames; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D011994:Recombinant Proteins