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Four nanostructured Au electrodes were prepared by a simple and templateless electrochemical deposition technique. After complexing with a biotinylated polythiophene derivative (PTBL), photocurrent generation and performance of PTBL/Au-nanostructured electrodes as photoelectrochemical biosensors were investigated. Among these four nanostructured Au electrodes, vertically oriented nanobranches on the electrode significantly improved the photoelectric conversion, because the vertically oriented nanostructures not only benefit light harvesting but also the transfer of the photogenerated charge carriers. Owing to this advantaged nanostructure, the PTBL/Au-nanobranch electrode showed higher sensitivity and faster response times in the photoelectrochemical detection of a streptavidin-biotin affinity reaction compared to a PTBL/Au-nanoparticle electrode.