Fig. 5: Nb14-32-Fc and its parent antibodies interfere with cell fusion and Nb14 interferes with gD binding to gH/gL.

a HEK-293T cells were co-transfected with glycoprotein expression plasmids (PLVX-gB-Zsgreen, PLVX-gD-Zsgreen, PLVX-gH-Zsgreen and PLVX-gL-Zsgreen) and cultured in medium containing 50 μg/ml antibodies at 37 °C for 48 h. Cells were stained with DAPI and observed under a fluorescence microscope. Scale bars, 500 μm. The relative Zsgreen area was quantified using ImageJ, with the fusion activity of the control set to 100%. b CHO cells which were co-transfected with glycoprotein expression plasmids and pG5-Luc were mixed with HEK-293T cells co-transfected with pACT-Myod and pBind-Id. Mixed cells were co-cultured in fresh media containing 50 μg/ml antibodies. After 48 h, cell-to-cell membrane fusion was evaluated using luciferase activity. The statistical significance in (a) and (b) was determined by comparing with the control group (no antibody group) using Dunnett’s multiple comparison tests in a one-way ANOVA analysis, revealing a significant difference with ****p < 0.0001, ***p < 0.001, **p < 0.01, *p < 0.05, p > 0.05(ns). ns, no significant. Data and error bars are mean ± S.D, n = 3 biological independent experiments. c Multi-concentration cell-based ELISA-binding assay of HSV-2 gD towards gH/gL expressed on CHO cells. The OD₄₅₀ values are depicted by curves. d The inhibition of Nb14-Fc activities was characterized by competitive ELISA, which demonstrated the ability of Nb14-Fc to inhibit the binding of HSV-2 gD to gH/gL expressed on CHO cells. Data and error bars in (c) and (d) are mean ± S.D, n = 3 biological independent experiments. Source data are provided as a Source Data file.