Characterization of biomembranes by spectral ellipsometry, surface plasmon resonance and interferometry with regard...

Phospholipid bilayers with transport proteins and antigen/antibody interfaces are considered to be suitable biosensor systems. The quality of such membranes or interfaces depends on the properties of the layers. Optical methods have proved to be an appropriate tool for characterizing those layers in situ and in a non-destructive manner. Two systems with potential for biosensor applications are characterized by some of these methods: phospholipid bilayer membranes spread from vesicle solution and protein-antigens both adsorbed on planar solid support. The results of spectral ellipsometry, surface plasmon resonance (SPR) and spectral interferometry are compared with respect to quality of characterization, expenditure of sample preparation and measurement, and time resolution. The phospholipid membranes adsorbed show a relatively low refractive index and a relatively high thickness. Bruggeman effective medium approximation is used to calculate the effective layer thickness. This result is compared to SPR measurements. A correlation between thickness and vesicle concentration may be detected. Further, the test protocol of an immunoassay is examined by spectral interferometry and SPR. Thicknesses determined are compared to results obtained by applying spectral ellipsometry. The data measured by ellipsometry are in agreement with the molecular dimensions of the immunoglobulins. Differences between details can be explained by physical considerations.