Reactivity of mAbs 3G1 and 2G4 to small nucleic acid oligonucleotides in capture ELISA.

<p>Purified mAb 2G4 was coated to 96-well plates at predetermined optimal concentration. MAb 3G1 was concentrated from hybridoma supernatant and coated at a predetermined optimal dilution. Capture of biotinylated oligonucleotides was measured by probing with streptavidin-HRP. A) Reactivity to biotinylated double-stranded RNA molecules (0.25–50 pmol/well) of varying lengths was tested in capture ELISA format. Molecule sizes tested were 21 bp, 30 bp, 40 bp, 50 bp. * denotes concentration at which difference between test samples and negative control (21 bp molecule) becomes significant. Statistical significance was analysed using two-way ANOVA, followed by Dunnett test for multiple comparisons (P<0.0001). B) Reactivity of mAbs to biotinylated nucleic acid molecules was tested in capture ELISA format using predetermined optimal concentrations (5–15 pmol/well). Molecules tested were dsDNA, ssDNA, ssRNA and RNA:DNA hybrid all 50 bp in length. **** denotes significant difference between test samples and the negative control (21 bp dsRNA). Statistical significance was analysed using one-way ANOVA, followed by Dunnett test for multiple comparisons (P<0.0001).