Since the discovery of polydopamine (PDA), there has been a lot of progress on using this substance to functionalize many different surfaces. However, little attention has been given to prepare functionalized surfaces for the preparation of flexible electrochemical paper-based devices. After fabricating the electrodes on paper substrates, we formed PDA on the surface of the working electrode using a chemical polymerization route. PDA nanofilms on carbon were characterized by contact angle (CA) experiments, X-ray photoelectron spectroscopy, scanning electron microscopy, atomic force microscopy (topography and electrical measurements) and electrochemical techniques. We observed that PDA introduces chemical functionalities (RNH2 and RC═O) that decrease the CA of the electrode. Moreover, PDA nanofilms did not block the heterogeneous electron transfer. In fact, we observed one of the highest standard heterogeneous rate constants (ks) for electrochemical paper-based electrodes (2.5 ± 0.1) × 10−3 cm s−1, which is an essential parameter to obtain larger currents. In addition, our results suggest that carbonyl functionalities are ascribed for the redox activity of the nanofilms. As a proof-of-concept, the electrooxidation of nicotinamide adenine dinucleotide showed remarkable features, such as, lower oxidation potential, electrocatalytic peak currents more than 30 times higher when compared to unmodified paper-based electrodes and electrocatalytic rate constant (kobs) of (8.2 ± 0.6) × 102 L mol−1 s−1.