Characterization of recombinant porcine reproductive and respiratory syndrome virus-1 (PRRSV-1) soluble Glycosylated Protein 3 (sGP3) specific porcine monoclonal antibodies (mAbs)

Serial dilutions of purified recombinant mAbs were assessed by enzyme-linked immunosorbent assay (ELISA) to determine antigen specificity (A). An irrelevant Nipah virus glycoprotein-specific recombinant porcine mAb (NiV mAb) was included as a negative control. Mean data from two technical replicates ± SD are shown. Recognition of native PRRSV antigen was assessed by intracellular staining of MARC-145 cells infected with PRRSV-1 Olot/91 (B) or PRRSV-2 VR-2332 (C). mAb binding was revealed by addition of a fluorescent secondary antibody and analysis using the Incucyte® SX5 Live-Cell Analysis System. Technical singlets are shown. Overlapping 16-mer linear peptides spanning the PRRSV-1 GP3 ectodomain were screened for mAb recognition by ELISA (D–I). In addition to the four GP3-specific recombinant mAbs and the irrelevant NiV mAb, polyclonal sera from pig 64 were included. Mean data from two technical replicates ± SD are shown.