E‑Beam Nanostructuring and Direct Click Biofunctionalization of Thiol–Ene Resist

Electron beam lithography (EBL) is of major importance for ultraminiaturized biohybrid system fabrication, as it allows combining biomolecular patterning and mechanical structure definition on the nanoscale. Existing methods are limited by multistep biomolecule immobilization procedures, harsh processing conditions that are harmful to sensitive biomolecules, or the structural properties of the resulting protein monolayers or hydrogel-based resists. This work introduces a thiol–ene EBL resist with chemically reactive thiol groups on its native surface that allow the direct and selective “click” immobilization of biomolecules under benign processing conditions. We constructed EBL structured features of size down to 20 nm, and direct functionalized the nanostructures with a sandwich of biotin and streptavidin. The facile combination of polymer nanostructuring with biomolecule immobilization enables mechanically robust biohybrid components of interest for nanoscale biomedical, electronic, photonic, and robotic applications.