This invention discloses a biomimetic nanofluidic sensing membrane, its preparation method, and its applications. The biomimetic nanofluidic sensing membrane comprises a top supporting
lipid bilayer membrane and a bottom
anodic aluminum oxide membrane with a macroporous structure. A portion of the supporting
lipid bilayer membrane corresponds to and communicates with some macropores in the
anodic aluminum oxide membrane. An
ion carrier is embedded in the supporting
lipid bilayer membrane, and its surface is modified with
cholesterol-modified aptamers and
polyethylene glycol. This invention utilizes the lateral fluidity of the supporting lipid
bilayer to achieve multivalent and efficient capture of aptamers. The natural antifouling properties of phospholipids and the steric hindrance of
polyethylene glycol synergistically suppress non-
specific adsorption, and the detection
signal is amplified by the
ion current rectification effect induced by the asymmetric structure. The
detection limit for TNF-α is as low as 0.219 pM, and with
machine learning, it can accurately distinguish six proteins. The preparation method is simple, and the detection sensitivity and specificity are high, showing broad application prospects in
early disease diagnosis and high-
throughput protein analysis.