INV-21060
Background
Nanopore sensing is a promising technology that detects and sequences biomacromolecules without depending on biomolecule modification, labeling, or surface immobilization. Rapid detection of biomacromolecules revolutionizes the field of proteomics and disease diagnosis. Nanopore technology employs nanoscale pores between two electrolytic fluid chambers with an impermeable membrane. As the voltage is applied to the chamber, an ion current occurs, and translocation of macromolecules through the pore causes changes in the ion current. Fluctuations of the ion current and the duration time of translocation give information about the size, concentration, and structure of biomolecules.
Though nanopore sensing has been successful in single-molecule DNA sequencing, challenges still remain in the sequencing of single-molecule proteins. In fact, the inherent complexity of proteins in charge, volume, hydrophobicity, and structure make them difficult to be transported and sequenced.
This invention introduces methods and systems to overcome the current limitations for transporting proteins through nanopore sensors.
Researchers at Northeastern have invented a method for the passage of full-length proteins through nanopores. This invention uses a combination of a chaotropic agent (a chemical that changes the hydration shell around the protein molecule) and an ultrastable bilayer membrane polymer to achieve the full passage of proteins through nanopores. The use of highly denaturing agents that are resisted by the stable membrane is unique to this invention since nanopore measurements are not typically possible under such conditions. Additionally, this invention is an enzyme-free method, so the enzyme lifetime would not be a limiting factor in protein translocation. Not relying on the protein charge also enables sequencing of uncharged proteins through this method.