Radioimmunoassay (RIA) and null ellipsometry are two common methods to quantify adsorbed proteins.
However, the accuracy of null ellipsometry with a constant protein refractive index (n 1.465, k 0) at l 632.8 nm
has this far not been explored. The present study compared the methods, and the degree of agreement between the
simplified single wavelength null ellipsometry and RIA to quantify adsorbed proteins was explored on different
surfaces. The quantification methods agreed well when A, ngstro¨m smooth hydrophilic or hydrophobic silicon surfaces,
and freshly radio-labelled proteins were used. Some discrepancies were noted when either rough surfaces or stored
and aged labelled proteins were used. The differences decreased when the aged protein solution was equilibrated with
freshly dissolved proteins at room temperature (RT) for a few hours prior to the surface incubations. Significant
differences were also noted between the methods when albumin was adsorbed at it’s iso-electric point (pH 4.8)
