An applicability of irradiation-promoted exchange reaction (IPER) to the fabrication of mixed self-assembled monolayers (SAMs) composed of the protein-repelling matrix and the moieties bearing binding sites for specific attachment of a target protein is demonstrated. As test systems, we took mixed films of oligoethylene glycol (OEG)Substituted alkanethiols (OEG-ATs) and biotin-substituted alkanethiols (BATs) on Au{1 1 1}. Such SAMs are suitable for the specific immobilization of avidin protein and its variants. The composition of the mixed OEG-AT/BAT SAMs could be precisely controlled by varying the irradiation dose, which is important prerequisite for the fabrication of the respective patterns by electron-beam lithography, While the general trend in the immobilization of avidin onto the mixed OEG-AT/BAT SAMs prepared by IPER was found to be consistent with the earlier reports regarding the analogous films fabricated by the coassembly method, the concentration of the BAT component in the mixed SAMs needed for the maximum surface coverage of the specific protein was found to be somewhat lower, and the maximum avidin coverage somewhat higher in the case of IPER as compared to the coassembly method. We ascribe these differences to the lack of phase segregation and better separation of the individual BAT species in the OEG-AT matrix in the case of IPER.