Nanobiosensor design utilizing a periplasmic E. coli receptor protein immobilized within Au/polycarbonate nanopores

A new type of nanopore sensor design is reported for a reagent-less electrochemical biosensor with no analyte "tagging" by fluorescent molecules, nanoparticles, or other species. This sensor design involves immobilization within Au-coated nanopores of bacterial periplasmic binding proteins (bPBP), which undergo a wide-amplitude, hinge-twist motion upon ligand binding. Ligand binding thus triggers a reduction in the effective thickness of the immobilized protein film, which is detected as an increase in electrolyte conductivity (decrease in impedance) through the nanopores. This new sensor design is demonstrated for glucose detection using a cysteine-tagged mutant (GGR Q26C) of the galactose/glucose receptor (GGR) protein from the bPBP family. The GGR Q26C protein is immobilized onto Au nanoislands that are deposited within the pores of commercially available nanoporous polycarbonate membranes.