In this work is reported the development and application of an amperometric biosensor for the determination of total biogenic amines content by using the commercial diamino oxidase (DAO from Porcine kidney E.C. 1.4.3.6) as the biocomponent, entrapped by glutaraldehyde onto an electrosynthesized bilayer film. In order to minimize both the fouling and the interference caused by the direct electrochemical oxidation of both the analytes (i.e., biogenic amines) and the common interferents usually present in food products the performances of Pt and Au electrodes and of several electroproduced anti-interferents mono- and bi-layer films were tested. In spite of a very low activity of the commercial DAO, the biosensor displayed a high response sensitivity in flow experiments, short response time, a good linear response and low detection limits. The excellent anti-interference characteristics allowed the use of the biosensor in screening analysis of food products.