Affinity determination of RAbDMvIII by Saturation ELISA. Calibration curve of the binding of RAbDMvIII and RAbvIII recombinant antibodies to coated EGFRvIII in the ELISA. A) The concentration range for RAbDMvIII and RAbvIII was 20 to 0.02 ug/ml (x axis) and the absorbance was read at 450 nm (Y axis). The assay was analyzed twice. B) Saturation ELISA shows the Scatchard plot of the binding of RAbDMvIII to EGFRvIII. The resulting data is plotted as ratio of α/c (y axis) as function of alpha (x axis) where α is the fraction of bound antibody sites and c is the concentration of free antigen. The slope is -K which is the affinity constant of antibody and that is 1.7 × 107 liters/mole (M-1).
Gupta et al. BMC Biotechnology 2010 10:72 doi:10.1186/1472-6750-10-72