This paper describes the use of new amphiphilic polyelectrolytes for protein immobilization. Monomolecular films of glucose oxidase (GO) and monoamine oxidase (MAO) stabilized by amphiphilic polyelectrolytes (polyethyleneimine and poly-4-vinylpyridine derivatives modified by lauryl chains) were formed on a water surface in a Langmuir-Blodgett trough. The compressed films containing the enzymes were transferred onto the surface of a polypropylene membrane of a Clark electrode according to the Languir-Schaefer method. The analytical responses of the resulting biosensors were linear over the range 1-10 mM of glucose and 8-100 mu M of tyramine. Furthermore, direct functional coupling of GO and ferrocenecarboxylic acid in multilayer films stabilized by amphiphilic polyelectrolytes was demonstrated. The amperometric response of such a sensor was linear over the range 1-20 mM of glucose. The dependence of the kinetic parameters of the enzymes on the amphiphilic polyelectrolyte structures is discussed.