Nanohybrid biosensor based on mussel-inspired electro-cross-linking of tannic acid capped gold nanoparticles and enzymes

Complementary tools to classical analytical methods, enzymatic biosensors are widely applied in medical diagnosis due to their high sensitivity, potential selectivity, and their possibility of miniaturization/automation. Among the different protocols of enzyme immobilization, the covalent binding and cross-linking of enzymes ensure the great stability of the developed biosensor. Obtained manually by drop-casting using a specific cross-linker, this immobilization process is not suitable for the specific functionalization of a single electrode out of a microelectrode array. In the present work, we developed a nanohybrid enzymatic biosensor with high sensitivity by a mussel-inspired electro-cross-linking process using a cheap and abundant natural molecule (tannic acid, TA), gold salt, and native enzymes. Based on the use of a cheap natural compound and gold salt, this electro-cross-linking process based on catechol/amine reaction (i) is versatile, likely to be applied on any kind of enzymes, (ii) does not require the synthesis of a specific cross-linker, (ii) gives enzymatic biosensors with high and very stable sensitivity over two weeks upon storage at room temperature and (iv) is temporally and spatially controlled, allowing the specific functionalization of a single electrode out of a microelectrode array. Besides the development of microbiosensors, this process can also be used for the design of enzymatic biofuel cells.