An electrical biosensor exploiting a nanostructured semiconductor is a promising technology for the highly sensitive, label-free detection of biomolecules via a straightforward electronic signal. The facile and scalable production of a nanopatterned electrical silicon biosensor by block copolymer (BCP) nanolithography is reported. A cost-effective and large-area nanofabrication, based on BCP self-assembly and single-step dry etching, is developed for the hexagonal nanohole patterning of thin silicon films. The resultant nanopatterned electrical channel modified with biotin molecules successfully detects the two proteins, streptavidin and avidin, down to nanoscale molarities (≈1 nm). The nanoscale pattern comparable to the Debye screening length and the large surface area of the three-dimensional silicon nanochannel enable excellent sensitivity and stability. A device simulation confirms that the nanopatterned structure used in this work is effective for biomolecule detection. This approach relying on the scalable self-assembly principle offers a high-throughput manufacturing process for clinical lab-on-a-chip diagnoses and relevant biomolecular studies.