GENERALIZABLE NANOPORE SENSOR FOR HIGHLY SPECIFIC PROTEIN DETECTION AT SINGLE-MOLECULE PRECISION

Smart overview of the Invention

A novel sensing element has been developed that combines a protein nanopore with a programmable antibody-mimetic binder. This modular architecture allows for the detection of a wide array of proteins while maintaining high specificity and sensitivity. The system was validated using diverse protein analytes, which produced distinct electrical signatures based on their identity and concentration at the nanopore's tip.

Need for Advanced Protein Detection

Accurate protein biomarker detection is critical for advancements in disease diagnostics and therapeutics. Despite progress in quantitative proteomics, many proteins remain unexplored due to the limitations of existing detection methods. Traditional nanopore sensors often require proteins to be confined within the nanopore, which can hinder their natural interactions and reduce detection efficacy. A more effective approach involves detecting proteins outside the nanopore, which necessitates innovative sensor designs.

Innovative Modular Sensor Features

The new sensor utilizes a monobody, derived from the fibronectin type III domain, fused to a tFhuA nanopore. This design not only preserves the sensor's structural integrity but also enhances its ability to generalize across various protein analytes by simply modifying the binding interface. The flexible tether connecting the binder to the nanopore allows for effective detection of proteins with varying characteristics in complex solutions.

Methodology for Protein Detection

The detection process involves reconstituting the engineered polypeptide chain within a lipid membrane under a set membrane potential. When exposed to a solution containing target proteins, any changes in membrane potential are measured, indicating successful binding events. This method enhances the dynamic range of detection and allows for real-time monitoring of protein interactions without steric hindrance from the nanopore.

Potential Applications and Impact

The developed nanopore sensors are expected to significantly influence the field of nanoproteomics by enabling direct measurement of protein binding events across diverse biochemical environments. Their ability to identify and quantify biomarkers in complex biofluids opens new avenues for research in disease progression and treatment strategies, making them a valuable tool in modern proteomics.