Unformatted text preview: Scatchard Plot : r= bound ligand / total antibody c= free ligand n= number of binding sites per antibody molecule Slope = -Ka X-intercept = n r/c = Ka(n) –Ka(r) Note: this only works if the antibody is homogeneous: all antigen binding sites identical, e.g. myeloma protein or a monoclonal antibody. Polyclonal antibodies will yield a curve due to different Ka’s for different isoforms. (2) Surface Plasmon Resonance Resonance units are proportional to the degree of binding of soluble ligand to the immobilized receptor. (or soluble antibody to immobilized antigen, as shown here ) - Determining the amount of binding at equilibrium with different known concentrations of receptor (antibody) and ligand (protein antigen) allows you to calculate equilibrium constants (Ka, Kd). -Rate of dissociation and association (k off , k on ) can also be calculated (1) Equilibrium Dialysis...
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This note was uploaded on 05/06/2008 for the course MCB 150 taught by Professor Coscey during the Spring '08 term at Berkeley.
- Spring '08