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US 12,735,712 B2Granted
Electrochemical detection nanostructure, systems and uses thereof
AssigneeAuburn University
InventorsChristopher J. Easley, Subramaniam Somasundaram
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Abstract
Described herein are DNA-nanostructures that can be used in an assay to detect and/or quantify an analyte of interest. Aspects of the DNA-nanostructure can include a single DNA molecule composed of hairpin structural motifs, an anchor recognition moiety, and a signal moiety, where the anchor recognition moiety and the signal moiety are in effective proximity to each other such that the tethered diffusion of the signal molecule can be altered based upon binding status of the anchor recognition moiety. Also described herein are methods of making and using the DNA-nanostructures.
Claims16
A method for detecting an analyte, the method comprising:
- contacting a sample containing the analyte to an anchor recognition moiety coupled to a DNA nanostructure, wherein the DNA nanostructure comprises:
- a single continuous DNA molecule comprising:
- a first hairpin structural motif, a second hairpin structural motif, a first segment of single stranded DNA, and a second segment of single stranded DNA, wherein the first hairpin structural motif and the second hairpin structural motif are attached to each other via the first segment of single stranded DNA, wherein the second segment of single stranded DNA is attached to the second hairpin structural motif such that the second segment of single stranded DNA forms a single stranded tether region at one end of the single continuous DNA molecule;
- the anchor recognition moiety, wherein the anchor recognition moiety is covalently coupled to a region of the single continuous DNA molecule between the first hairpin structural motif and the second hairpin structural motif;
- and
- a signal moiety coupled to an end of the single continuous DNA molecule opposite from the tether region, wherein the signal moiety and the anchor recognition moiety on the single continuous DNA molecule are in effective proximity to each other such that binding of the analyte to the anchor recognition moiety changes a tethered diffusion rate of the signal moiety,
- wherein a terminal base of the second segment of single stranded DNA of the DNA nanostructure is coupled to a surface of an electrode;
- and
- detecting a change in the tethered diffusion rate of the signal moiety relative to the surface of the electrode when the analyte binds to the anchor recognition moiety wherein said detecting the change in the tethered diffusion rate of the signal moiety comprises detecting a change in an electrochemical current at the surface of the electrode.
6 more elements
A method for detecting an analyte, the method comprising:
- contacting a sample containing the analyte to an anchor recognition moiety coupled to a DNA nanostructure, wherein the DNA nanostructure comprises:
- a single continuous DNA molecule comprising:
- a first hairpin structural motif, a second hairpin structural motif, a first segment of single stranded DNA, and a second segment of single stranded DNA, wherein the first hairpin structural motif and the second hairpin structural motif are attached to each other via the first segment of single stranded DNA, wherein the second segment of single stranded DNA is attached to the second hairpin structural motif such that the second segment of single stranded DNA forms a single-stranded tether region at one end of the single continuous DNA molecule;
- the anchor recognition moiety, wherein the anchor recognition moiety is covalently coupled to the single continuous DNA molecule and extends from the single continuous DNA molecule;
- and
- a signal moiety coupled to an end of the single continuous DNA molecule opposite from the tether region, wherein the signal moiety and the anchor recognition moiety on the single continuous DNA molecule are in effective proximity to each other such that binding of the analyte to the anchor recognition moiety changes a tethered diffusion rate of the signal moiety,
- wherein a terminal base of the second segment of single stranded DNA of the DNA nanostructure is coupled to the surface of the electrode;
- and
- detecting a change in the tethered diffusion rate of the signal moiety relative to the surface of the electrode when the analyte binds to the anchor recognition moiety wherein said detecting the change in the tethered diffusion rate of the signal moiety comprises detecting the change in an electrochemical current at the surface of the electrode.
6 more elements
A method for detecting an analyte, the method comprising:
- contacting a sample containing the analyte to an anchor recognition moiety coupled to a DNA nanostructure, wherein the DNA nanostructure comprises:
- a single continuous DNA molecule comprising:
- a first hairpin structural motif, a second hairpin structural motif, a first segment of single stranded DNA, and a second segment of single stranded DNA, wherein the first hairpin structural motif and the second hairpin structural motif are attached to each other via the first segment of single stranded DNA, wherein the second segment of single stranded DNA is attached to the second hairpin structural motif such that the second segment of single stranded DNA forms a single-stranded tether region at one end of the single continuous DNA molecule;
- the anchor recognition moiety, wherein the anchor recognition moiety is covalently coupled to the single continuous DNA molecule and extends from the single continuous DNA molecule;
- and
- a signal moiety, wherein the signal moiety is coupled to an end of the single continuous DNA molecule opposite from the tether region, wherein the signal moiety and the anchor recognition moiety on the single continuous DNA molecule is in effective proximity to each other, wherein the DNA nanostructure is coupled to the surface of the electrode by a terminal base of the second segment of single stranded DNA of the DNA nanostructure such that binding of the analyte to the anchor recognition moiety changes a tethered diffusion rate of the signal moiety and a change in the tethered diffusion rate of the signal moiety changes an electrochemical current at the surface of the electrode;
- and
- detecting a change in the tethered diffusion rate of the signal moiety relative to the surface of the electrode when the analyte binds to the anchor recognition moiety wherein said detecting the change in the tethered diffusion rate of the signal moiety comprises detecting the change in an electrochemical current at the surface of the electrode.
5 more elements
Dependencies are inferred from the claim text.