The formation of stable supramolecular interactions between biotin and beta-cyclodextrin was studied. An association constant of 3 X 10(2) M-1 could be determined by NMR measurements by mapping the high field shift differences of the beta-cyclodextrin protons (H-3) at different biotin concentrations. With the aim to demonstrate a new alternative for the immobilization of bioreceptors, biotin and beta-cyclodextrin tagged biomolecules were immobilized on transducer surfaces, which were functionalized with the correspondent hostguest partner. The reliability of this new affinity system was investigated using two enzymes (glucose oxidase and polyphenol oxidase) as biomolecule models. This supramolecular inclusion complex shows clear advantages to the classic biotin-(strept)avidinbiotin system due to a detrimental effect of the additional avidin layer reducing the transduction efficiency. A 7-fold increase in the maximum current density and an almost 20 times higher sensitivity were exhibited by the immobilized biological layer obtained using this new hostguest system