Continuous liquid-phase piezoelectric biosensor for kinetic immunoassays

The reactions of immunoglobulins with protein A and, subsequently, of antibodies to these immunoglobulins were continuously monitored by a piezoelectric biosensor. AT-cut crystals, with a fundamental resonant frequency of 10 MHz, were mounted in a Plexiglas holder, and one side was directly coated with protein A. Upon exposure to solutions containing rabbit or human IgG the resonant frequency was followed continuously and its decrease due to the binding of IgG to protein A observed. Subsequent addition of sheep antihuman IgG to the now immobilized human IgG caused a specific 3-fold further decrease in resonant frequency. We observed a frequency change of approximately 1 Hz fore each 10 ng of added immunoglobulin. Decreasing the pH to 3 released the bound IgG but not the protein A and permitted reuse of the crystal for further IgG binding.