Figure 5.

Binding of different concentrations of (a) sCD4 and (b) mAb 17b to E275C and the NeutrAvidin-captured AE21–E275C covalent conjugate, after reference subtraction and normalization to control for the amount of immobilized protein (Materials and Methods). The schematics on the left side of each panel depict the assay configuration. E275C was immobilized on the surface using standard amine coupling (Materials and Methods). The AE21–E275C covalent conjugate was captured from a filtered reaction mixture using surface-immobilized NeutrAvidin via the biotin handle on the peptide. Different concentrations of analytes were passed over the surfaces at a rate of 100 μL/min. Response levels at the end of the association phase (80 s) were used as the representative equilibrium signal at each concentration.