Nanopores with an Engineered Selective Entropic Gate Detect Proteins at Nanomolar Concentration in Complex Biological Sample

Biological nanopores enable the electrical detection of biomolecules, making them ideal sensors for use in health-monitoring devices. Proteins are widely recognized as biomarkers for various diseases, but they present a unique challenge due to their vast diversity and concentration range in biological samples. Here, inspired by the nuclear pore complex, we incorporated a layer of disordered polypeptides into the biological nanopore YaxAB. This polypeptide mesh formed an entropic gate, significantly reducing the entry of proteins from a highly concentrated mixture, including blood. The introduction of a specific recognition element within the disordered polypeptides allowed targeted proteins to penetrate through the nanopores, where they were recognized by specific current signatures. This biosensing approach allowed for the recognition of nanomolar proteins directly from blood samples without prior sample preparation. This work paves the way for the next generation of nanopore sensors for the real-time detection of proteins in blood.