Analysis of active antibody concentration. Separation of affinity and concentration parameters

Antibody binding to surfaces with differing amounts of immobilised antigen was measured in a biosensor system using surface plasmon resonance detection. Binding rates obtained during the initial binding phase on high density antigen surfaces were proportional to antibody concentration and independent of antigen-antibody affinity. One antibody calibration curve covering the range from 0.5 to 160 nM (0.08-25 micrograms/ml) antibody was valid for IgG antibodies with different antigen specificities. To illustrate the use of this methodology active antibody concentrations were analysed in culture media and in rabbit serum.