The structural basis of protective and nonprotective human monoclonal antibodies targeting the parainfluenza virus type 3 hemagglutinin-neuraminidase

Figure 2. Fusion inhibition of mAbs from each site was assessed by transfecting Vero-SLAM cells with either HN or F and GFP 3 hrs prior to adding mAb. Syncytia formation was visualized 24 hours following transfection and fusion inhibition was evaluated by comparison to the PBS-treated control. Previously identified anti-F mAb PIA174 was used as a positive control.<br>Figure S4. Raw Size exclusion chromatography data for the curves in Figure S4. PIV3 HN eluted at approximately 14-15 mL on the Superdex S200, 10/300 column. PIV3HN-05 Fab eluted at approximately 15 mL on the Superdex S200, 10/300 column. PIV3 HN complexed with PIV3HN-13 Fab eluted at approximately 12 mL, and excess Fab eluted at approximately 15 mL on the Superdex S200, 16/600 column. (D) PIV3 HN complexed with PIV3HN-05 Fab and PIV3HN-13 Fab eluted at approximately 11 mL, and excess Fab eluted at approximately 15 mL. PIV3 infected cells mAb staining. Raw flow cytometry data from Figure 1D. PIV3-infected LLC-MK2 cells were stained with anti-PIV3 HN mAbs PIV3HN-05 and PIV3HN-09 conjugated to PE. Anti-F mAb PIA174 was used as a positive control, and a mAb against human metapneumovirus, MPV467, was used as a negative control. Both anti-HN mAbs and anti-F mAbs were detected on virally infected cells. Data in the manuscript was analyzed in FlowJo.<br><br>