Harvesting the spatial resolution and positioning accuracy of nanopatterning technology with the programmability and precision of biomacromolecules, has the potential to enable new technologies for fundamental biophysical research and clinical bioelectronic devices. These devices such as biosensors, disease diagnostic and treatment implants, benefit from well-defined surface nanopatterns, offering superior single-molecule resolution and sensitivity compared to homogeneous or micropatterned surfaces. For example, optical nanostructures such as zero-mode waveguides with selective immobilization of enzyme molecules in the confined sub-diffraction observation volume enable single-molecule investigation of enzymatic activities at biologically relevant concentrations. Nanoscale features on material surfaces (topographical and chemical cues) can program cell adhesion and fate, offering fundamental biological insights into cell behavior in response to their microenvironment. These insights are crucial for advancing cell culture materials and regenerative medicines including implants and stem cell therapeutics. The successful development of bioelectronic or optoelectronic devices interfacing with biological systems with single-molecule resolution will require precise design and customization of surface modification layers with exact nanoscale patterning at the inorganic/bio interface. These surface modification layers which consist of molecular coatings in direct contact with biological systems, enable the viability of the interface by fulfilling various functions. They can adjust surface hydrophilicity or hydrophobicity, facilitate molecular immobilization or antifouling in specific regions, regulate short-range interactions, and even provide specific molecular recognition.