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Binary operating (YES/NO) biosensors based on unconventional biocomputing concept represent a novel approach to the analysis of various biomarkers and environmental signals. The binary biosensors with implemented Boolean logic process analyte input signals using chemical means without electronic computers. They can be used for extension of the analytical output signals to chemical actuation and can be integrated with wearable analytical devices. The progress in this novel biosensing approach is strongly dependent on achievements in biomolecular computing, particularly based on enzyme-catalyzed reactions. The easy interpretation of the analytical results represented in the binary YES/NO form allows convenient adaptation of the biosensors for end-user and point-ofcare within various biomedical and bioelectronic applications.