Polyaniline-based hybrid membrane for single-molecule protein nanopore analysis under high voltage

In vitro reconstituted lipid bilayers are the key to the birth of bio-nanopore technology, although they are fragile and inadequately stable for long-term monitoring applications. Amphiphilic copolymers with high robustness have been developed to circumvent this issue. However, it remains a major challenge to achieve the required fluidity for protein insertion. Here we engineered a polyaniline-based hybrid membrane (PANIM) that exhibits the stability needed for fabricating high-throughput biosensors. This enhanced durability enables prolonged operation without compromising the functional properties of embedded α-hemolysin (α-HL) nanopores. Based on this, we demonstrate high voltage analysis for G-triplex conformations through α-HL in PANIM, which facilitates in-depth study of molecular conformations and provides a novel platform that has great potential for nanopore sensing.