High throughput stochastic bio-molecular sensor

A method for identifying a target signature of and quantifying the abundance of one or more targets in a sample, comprising:

• utilizing a device that comprises a bioprotonic conducting material forming a planar array of protodes on a non-conducting substrate, wherein each protode in the planar array of protodes is electrically isolated and individually addressable;
• a plurality of DNA origami tethered DNA nanopores immobilized to the planar array of protodes, wherein the outer surface of each of the plurality of DNA origami tethered DNA nanopores comprises one or more hydrophobic moieties;
• an insulating membrane, defining a space inferior to the membrane and a space superior to the membrane, wherein each of the DNA origami tethered DNA nanopores spans the membrane and provides an ionic pathway between the bioprotonic conducting material inferior to the membrane and bulk solution;
• a power supply in electrical contact with each protode to provide an electric potential difference across the membrane;
• and
• a detector, configured to independently acquire and resolve signals in real-time from each protode in the planar array of protodes, to detect changes in electrical signal through the DNA origami tethered DNA nanopore over time as each DNA origami tethered DNA nanopore interacts with the one or more targets, wherein the changes in electrical signal can comprise changes in signal magnitude, signal duration, or frequency of signal changes, and wherein signals from individual protodes are independently resolved to discriminate one or more targets in the sample, executing the steps of:
• introducing a sample into the space superior to the membrane of two or more DNA origami tethered DNA nanopores immobilized to the planar array of protodes;
• applying an electric field across the membranes;
• monitoring changes in signal magnitude, signal duration and frequency of signal change across the membranes to define one or more target signatures;
• and
• determining a presence and abundance of a target in the sample as a function of the one or more target signatures.

A method for fabricating a stochastic sensing device for sensing one or more targets in a sample, comprising:

• depositing a bioprotonic conducting material onto a non-conducting material arranged to form one or more protode contact areas;
• electrically isolating each protode contact area;
• introducing DNA origami to the protode contact areas for self-assembly onto the bioprotonic conducting material,
• wherein the DNA origami comprises one or more anchoring linkers;
• introducing DNA nanopores to the protode contact areas,
• wherein the DNA nanopores comprise one or more nanopore anchoring linkers complementary to the anchoring linkers on the DNA origami, one or more hydrophobic moieties on an outer surface of the DNA nanopores, and one or more target binding moieties opposite the nanopore anchoring linkers;
• wherein the nanopore anchoring linkers bind to the anchoring linkers on the DNA origami thereby immobilizing the nanopores on the DNA origami;
• and
• wherein the target binding moieties interact weakly with one or more determinants on the target;
• forming a membrane within the electrically isolated protode contact area and surrounding the DNA nanopore, defining a space inferior to the membrane and a space superior to the membrane, such that one ionic pathway exists between the bioprotonic conducting material and the space superior to the membrane, through the nanopore;
• wherein each protode contact area contains on average one immobilized DNA nanopore;
• and
• wherein the stochastic sensing device can detect changes in signal magnitude, signal duration and frequency of signal change across the membrane upon exposure to one or more targets in a sample.