Refractive index and layer thickness of an adsorbing protein as reporters of monolayer formation

A method is presented for a separate real-time determination of refractive index and layer thickness of an adsorbing thin layer. The changing angular deflections of TE and TM modes in a dedicated planar waveguide structure are measured. A resolution of 0.01 in the refractive index and 0.5 nm in the average thickness is obtained. The method is illustrated with experimental results on the binding of an antibody to the substrate, both in a physisorption and in an immunoreaction. In the latter, results are consistent with an end-on binding of the antibody to the surface.