Multilayered assemblies of glucose oxidase (GOD) and poly(4-vinylpyridine) complex of osmium were fabricated successfully by alternate deposition technique. Ultraviolet-visible spectra showed a continuous deposition process of GOD and the polymer on quartz slides. X-ray diffraction indicated a process of layer-by-layer deposition of GOD and the polycation. Cyclic voltammetry indicated that the electrode fabricated in this way had the function of oxidation of glucose, in which osmium transferred electrons successfully between the immobilized GOD and the electrode surface.