Figure 4.
Determination of antigen dissociation constants (Kd) of c11 and c11L34Ser scFvs. Purified proteins c11 (2 nM) and c11L34Ser (1 nM) were mixed with different concentration of GCN4(7P14P) peptide and incubated for 1 hour before analysis. Samples were injected over the sensor chip coated with BSA-GCN4(7P14P) conjugate. From the linear sensograms, the slopes (resonance units vs. time in sec) were plotted against the corresponding total soluble antigen concentration. The slopes correlate to uncomplexed scFv in the injected solutions. From a fit with Eq. 1, Kd was calculated. Each point is the average of four independent experiments.