The PhD thesis is focused on the study of affinity interactions of biomolecules on a solid surface by TSM acoustic methods for the purpose of biosensor development. TSM is a very sensitive method, because even small changes in contact with molecules on the surface and interface layer of biologically active layer affect influence on the overall acoustic properties of the sensor. In the case of biosensors, small changes in mass of the QCM crystal layer cause significant changes in the detected signals such as resonant frequency and motional resistance. Atomic force microscopy (AFM) was used for three-dimensional surface imaging. AFM is the first universal tool which allows us to explore the nanoworld and therefore there is no doubt that it is widely used in various scientific and technical areas, including basic as well as applied research. Current research in the design of biosensors is focused on the use of different types of nanostructures. For the detection of dopamine we used nanostructure 25,26,27,28 tetrakis(11-sulfanylundecyloxy)calix[4]arene (CX4). We have designed three types of chemically sensitive interfaces. Interface with CX4 and CX4 mixed with alkanthiols, hexadecanthiol (HDT) and dodecanthiol (DDT). For detection of cytochrome c, we used a synthetic receptor 37,38,39,40,41,42-hexakis(karboxymetoxy)-5,11,17,23,29,35-hexakis(1,1,3,3,-tetrametylbutyl)calix[6]arene (CX6). CX6 which was incorporated into the lipid membrane of DMPC. Thrombin was detected using a synthetically prepared single-stranded DNA -aptamers, which were sensitive to fibrinogen exosite on the thrombin molecule. We used single-stranded aptamer Biofibri-TT or a mixture of aptamer Biofibri-TT + Fibri-AT2 (homodimer). Immobilization of the aptamers on the sensor surface is very important, and therefore we tested two methods of aptamer immobilization. The first way consisting in using neutravidin, which was attached to the sensor surface and on the neutravidin was immobilized aptamer (or homodimer). As a second method of immobilization, we used DNA nanostructure DS3BT1-tetrahedron, which is used for oriented immobilization of neutravidin and aptamers.