Figure 6
From: Functional recombinant MHC class II molecules and high-throughput peptide-binding assays
ELISA-based peptide-MHC class II binding assay. Urea denatured α and β chains of DR1, DR2 and DR4 were diluted into titrations of CLIP or HA306–318 peptides in refolding buffer pH 8 (final concentrations of α and β chains were 31 and 2; 31 and 1; and 31 and 1 nM, respectively). After 24 h incubation at 18°C, the assay was developed as described in materials and methods. The measurements were background subtracted and analysed in Graphpad Prism as previously described. (A) Saturation curve for (black triangle) HA306–318 and (black square) CLIP binding to DR1 (left hand panel), DR2 (middle panel) or DR4 (right hand panel) plotted as Absorbance at 450 nm versus the logarithm(10) of the peptide concentration in nM. (B) EC50 values and signal to noise ratios (calculated as the ratio between signal at maximum peptide concentration and background). Not determined (N.D.).