Extended Data Fig. 1: Construction and characterization of m1A6, h1A6 and h1A6.2-LALA.
a, The quantity of cell-bound virus RNA was determined by qRT-PCR in L929SCARB2 cell-based inhibition assay. The values (n = 3) are expressed as mean ± SD of three experiments. Significance was determined using an unpaired, two-sided Student’s t test. Symbols: ***p < 0.001, ****p < 0.0001, n.s., not significant. b, Representative fluorescence confocal images of m1A6 against EV71- or CVA16-infected RD cells. The secondary antibody (green) was Alexa Fluor 488-conjugated goat anti-mouse. The nuclei were stained with DAPI (blue). Scale bar, 25 μm. Experiments were repeated twice, and one representative result is shown. c, Western blot analysis from an experment of m1A6 against EV71-B3, B4, C2, C4 sub-genotypes and four CVA16-B1b strains. d, In vivo animal protective efficacies of six versions of humanized 1A6 (h1A6). One-day-old mice (n = 5-8) were first challenged with a lethal dose of EV71-pSVA-MP4 or CVA16-190, and then treated with 1 mg/kg of one of the h1A6 antibodies, respectively, at 24 h post-infection. Mice were monitored daily for survival after inoculation. Experiments were repeated twice, and one representative result is shown. Survival curves were compared by the log-rank (Mantel-Cox) test. Symbols: *p < 0.05, **p < 0.01, ***p < 0.001, n.s., not significant. e, SDS-PAGE analysis from an experment of h1A6.2 and h1A6.2-LALA under reducing conditions. Lane M, protein markers; Lane 1, h1A6.2; Lane 2, h1A6.2-LALA. f and g, Binding efficacies of h1A6.2 and h1A6.2-LALA to EV71 (f) or CVA16 (g) recombinant VP2 protein evaluated with binding ELISA. The values (n = 3) are expressed as mean ± SD of three experiments. EC50 values were calculated with curves generated by nonlinear regression fitted.
