Fig. 1: Nucleic acid specificity and dsRNA length requirements of J2 antibody.
From: Structural basis of double-stranded RNA recognition by the J2 monoclonal antibody
a Sequences of single-stranded (ss) and double-stranded (ds) nucleic acids used in J2 binding analyses. RNA strands are shown in red; DNA in black. b SEC-MALS analysis of J2 Fab binding to a 27-bp dsRNA. c BLI sensorgrams using J2 IgG as immobilized ligand and the nucleic acids in (a) as analytes, at 400 mM KCl. d Plot of sensor responses (nm) against nucleic acid concentrations in BLI steady-state analysis. e dsRNAs of various lengths used for J2 binding analyses. f Bimolecular fluorescence polarization (FP) analysis of J2 IgG binding to a 3′-FAM-labeled Adenovirus VA-I RNA. ΔFP: change in FP expressed in dimensionless millipolarization (mP) units. Experiments were performed twice. g Competition FP analysis of dsRNAs in (e) in displacing 3′-FAM labeled VA-I RNA prebound to J2 IgG. Apparent IC50s are indicated. h IC50s derived from FP in (g) (upper) and maximum BLI sensor responses from (i) (lower). i BLI sensorgrams of dsRNAs in (e) binding to immobilized J2 IgG. Due to significant dependency of J2 binding on ionic strengths, 100 mM KCl was used for 6, 10, 14, and 20 bp; 200 mM KCl for 30 bp; 400 mM KCl for 40 and 50 bp dsRNA. j Steady-state analyses of BLI sensorgrams in (i). Apparent Kds are indicated. Values are mean ± s.d. of three biologically independent samples unless otherwise indicated.
