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A New Class of Nanopore Sensors for Rapid Detection of Bioanalytes
Protein nanopores have been used to detect small molecule analytes by monitoring changes in ionic current upon analyte binding in the nanopore lumen. The fixed inner diameter of the nanopore lumen, however, presents challenges for detection of proteins and other large bioanalytes. This invention provides a new class of protein nanopore sensors capable of detecting large bioanalytes without the need for such bioanalytes to enter the nanopore lumen. The nanopore is genetically engineered or chemically modified to contain a target-binding ligand in a flexible loop region at one end of the nanopore lumen. The dynamic movement of the flexible loop creates a distinct grating pattern when ionic current passes through the nanopore lumen. The nanopore-target analyte Interactions outside the pore lumen result in an instantaneous change in the gating pattern of the flexible loop, enabling rapid detection of the target analyte. This invention not only provides the ability to distinguish between protein homologues within an analyte mixture but also allows for the detection of target proteins in the presence of serum.
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