US-12625130-B2 - Nanopore sensor devices

US12625130B2US 12625130 B2US12625130 B2US 12625130B2US-12625130-B2

Abstract

An example of a nanopore sensor device includes one or more cis wells; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a lipid/solid-state membrane having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; a first concentration of an electrolyte within the one or more cis wells; and a second concentration of the electrolyte within the trans wells, wherein the first concentration is higher than the second concentration.

Inventors

Boyan Boyanov
Jeffrey G. Mandell
Seth M. McDonald

Assignees

ILLUMINA, INC.

Dates

Publication Date: 20260512
Application Date: 20220301

Claims (20)

1 . A nanopore sensor device, comprising: one or more cis wells; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a membrane selected from the group consisting of a lipid bilayer, a solid-state membrane, and a liquid film material, and having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; a first concentration of an electrolyte within the one or more cis wells; a second concentration of the electrolyte within the trans wells, wherein the first concentration is higher than the second concentration; a stimulus source coupled to each of the plurality of trans electrodes either individually or via multiplexing, wherein the stimulus source is to cause current to flow through the nanopore; and a controller coupled to the stimulus source, the controller configured to individually/selectively address the plurality of trans electrodes to cause an ionic current through the nanopore of an addressed trans electrode of the plurality of trans wells; wherein the ionic current comprises an amount of anions of the electrolyte translocating through the nanopore to the addressed trans well that is higher than an amount of cations of the electrolyte translocating through the nanopore from the addressed trans well.
2 . The nanopore sensor device as defined in claim 1 , wherein the controller is further configured to cause the stimulus source to apply a unipolar electric current between the cis electrode and the addressed trans electrode of the addressed trans well of the plurality of trans wells.
3 . The nanopore sensor device as defined in claim 1 , wherein the nanopore has a plurality of positively charged residues on an inner surface of the nanopore.
4 . The nanopore sensor device as defined in claim 3 , wherein the plurality of positively charged residues on the inner surface are located at a constriction zone of the nanopore.
5 . The nanopore sensor device as defined in claim 1 , wherein: the membrane is the lipid bilayer, and the lipid bilayer includes two opposing layers of phospholipids; or the membrane is the solid-state membrane, and the solid-state membrane is selected from the group consisting of silicon nitride, aluminum oxide, hafnium oxide, tantalum pentoxide, silicon oxide, polyamide, polytetrafluoroethylene, a two-component addition-cure silicone rubber, glass, and graphene; or the liquid film material is a diblock copolymer or a triblock copolymer.
6 . A nanopore sensor device, comprising: one or more cis wells; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a membrane selected from the group consisting of a lipid bilayer, a solid-state membrane, and a liquid film material, and having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; a first concentration of an electrolyte within the one or more cis wells; and a second concentration of the electrolyte within the trans wells, wherein the first concentration is higher than the second concentration; wherein a ratio of the first concentration to the second concentration ranges from about 10:1 to about 3:1.
7 . The nanopore sensor device as defined in claim 6 , further comprising: a stimulus source coupled to each of the plurality of trans electrodes either individually or via multiplexing, wherein the stimulus source is to cause current to flow through the nanopore; and a controller coupled to the stimulus source, the controller configured to individually/selectively address the plurality of trans electrodes to cause an ionic current through the nanopore of an addressed trans electrode of the plurality of trans wells.
8 . The nanopore sensor device as defined in claim 7 , wherein the controller is further configured to cause the stimulus source to apply a unipolar electric current between the cis electrode and the addressed trans electrode of the addressed trans well of the plurality of trans wells.
9 . The nanopore sensor device as defined in claim 6 , wherein the nanopore has a plurality of positively charged residues on an inner surface of the nanopore.
10 . The nanopore sensor device as defined in claim 9 , wherein the plurality of positively charged residues on the inner surface are located at a constriction zone of the nanopore.
11 . A nanopore sensor kit, comprising: a nanopore sensor device, including: one or more cis wells including a fluid inlet; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a membrane selected from the group consisting of a lipid bilayer, a solid-state membrane, and a liquid film material, and having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; and a first concentration of an electrolyte within the one or more cis wells and the plurality of trans well; a second concentration of the electrolyte to be introduced into the one or more cis wells through the fluid inlet such that the one or more cis wells contain the second concentration of the electrolyte and the plurality of trans wells contain the first concentration of the electrolyte at an initial cycle of the nanopore sensor device, wherein the second concentration is higher than the first concentration; a stimulus source coupled to each of the plurality of trans electrodes either individually or via multiplexing, wherein the stimulus source is to cause current to flow through the nanopore; and a controller coupled to the stimulus source, the controller configured to individually/selectively address the plurality of trans electrodes to cause an ionic current through the nanopore of an addressed trans electrode of the plurality of trans wells; wherein the ionic current comprises an amount of anions of the electrolyte translocating through the nanopore to the addressed trans well that is higher than an amount of cations of the electrolyte translocating through the nanopore from the addressed trans well.
12 . A method of detecting an ionic current to analyze a biological compound, comprising: providing a solid-state nanopore within a membrane separating a cis well and a trans well, the solid-state nanopore having a plurality of positively charged residues on an inner surface, and the plurality of positively charged residues being selected from the group consisting of an organic positively charged species, an inorganic positively charged species, or both organic positively charged species and inorganic positively charged species; providing an electrolyte within the cis well and the trans well; and applying an electric current between a cis cathode at least partially exposed to the cis well and a trans anode at least partially exposed to the trans well to generate an ionic current through the nanopore, wherein the plurality of positively charged residues of the nanopore inhibits translocation of cations from the trans well to the cis well during application of the electric current.
13 . The method as defined in claim 12 , wherein the plurality of positively charged residues on the inner surface are located at a constriction zone of the nanopore.
14 . The method as defined in claim 12 , wherein the applied electric current is a unipolar current.
15 . The method as defined in claim 12 , wherein the cis well comprises a higher concentration of the electrolyte than the trans well during application of the electric current.
16 . A method of detecting an ionic current to analyze a biological compound, comprising: providing a nanopore within a membrane separating a cis well and a trans well, the nanopore having a plurality of positively charged residues on an inner surface; providing an electrolyte within the cis well and the trans well; and applying an electric current between a cis cathode at least partially exposed to the cis well and a trans anode at least partially exposed to the trans well to generate an ionic current through the nanopore, wherein the plurality of positively charged residues of the nanopore inhibits translocation of cations from the trans well to the cis well during application of the electric current; and wherein one of: the electrolyte is a redox couple having a negative charge and is incorporated into a redox-inactive buffer that includes an anion having a diameter greater than a diameter of a constriction zone of the nanopore; and the electrolyte is a redox couple having a positive charge and is incorporated into a redox-inactive buffer that includes a cation having a diameter greater than a diameter of a constriction zone of the nanopore.
17 . A nanopore sensor device, comprising: one or more cis wells; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a membrane selected from the group consisting of a lipid bilayer, a solid-state membrane, and a liquid film material, and having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; an electrolyte solution including: a redox-inactive buffer that includes a redox inactive species having a diameter greater than a diameter of a constriction zone of the nanopore; and a redox couple.
18 . The nanopore sensor device as defined in claim 17 , further comprising: a stimulus source coupled to each of the plurality of trans electrodes either individually or via multiplexing, wherein the stimulus source is to cause current to flow through the nanopore; and a controller coupled to the stimulus source, the controller configured to individually/selectively address the plurality of trans electrodes to cause an ionic current through the nanopore of an addressed trans electrode of the plurality of trans wells.
19 . The nanopore sensor device as defined in claim 18 , wherein the ionic current comprises an amount of the redox couple translocating through the nanopore to the addressed trans electrode without an amount of the redox-inactive species.
20 . The nanopore sensor device as defined in claim 18 , wherein the controller is further configured to apply a unipolar electric current between the cis electrode and the addressed trans electrode of the addressed trans well of the plurality of trans wells.

Description

CROSS-REFERENCE TO RELATED APPLICATION This application claims the benefit of U.S. Provisional Application Ser. No. 63/168,646, filed Mar. 31, 2021, the contents of which is incorporated by reference herein in its entirety. BACKGROUND Various polynucleotide sequencing techniques involve performing a large number of controlled reactions on local support surfaces or within predefined reaction chambers. The designated reactions may then be observed or detected, and subsequent analysis may help identify or reveal properties of the polynucleotide involved in the reaction. Another polynucleotide sequencing technique has been developed that utilizes a nanopore, which can provide a channel for an ionic electrical current. A polynucleotide or label/tab of an incorporated nucleotide is driven into the nanopore, changing the resistivity of the nanopore. Each nucleotide (or series of nucleotides) or each label/tab (or series of labels/tags) yields a characteristic electrical signal, and the record of the signal levels corresponds to the sequence of the polynucleotide. In prior nanopore sensor devices (at t=0), the current is equally carried by the electrolyte translocating through the nanopore in opposite directions between a cis well and a trans well. However, such nanopore sequencing devices suffer from low lifetimes. SUMMARY In a first example, a nanopore sensor device comprises one or more cis wells; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a lipid/polymer/solid-state membrane having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; a first concentration of an electrolyte within the one or more cis wells; and a second concentration of the electrolyte within the trans wells, wherein the first concentration is higher than the second concentration. In a second example, a nanopore sensor kit comprises i) a nanopore sensor device, including: one or more cis wells including a fluid inlet; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a lipid/polymer/solid-state membrane having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; and a first concentration of an electrolyte within the one or more cis wells and the plurality of trans well; and ii) a second concentration of the electrolyte to be introduced into the one or more cis wells through the fluid inlet such that the one or more cis wells contain the second concentration of the electrolyte and the plurality of trans wells contain the first concentration of the electrolyte at an initial cycle of the nanopore sensor device, wherein the second concentration is higher than the first concentration. In a third example, a method of detecting an ionic current to analyze a biological compound comprises providing a nanopore within a membrane separating a cis well and a trans well, the nanopore having a plurality of positively charged residues on an inner surface of the nanopore; providing an electrolyte within the cis well and the trans well; and applying an electric current between a cis cathode at least partially exposed to the cis well and a trans anode at least partially exposed to the trans well to generate an ionic current through the nanopore, wherein the plurality of positively charged residues of the nanopore inhibits translocation of cations from the trans well to the cis well during application of the electric current. In a fourth example, a nanopore sensor device comprises one or more cis wells; a cis electrode; a plurality of trans wells, each of the plurality of trans wells separated from the one or more cis wells by a lipid/polymer/solid-state membrane having a nanopore; a plurality of trans electrodes, each of the plurality of trans electrodes associated with one of the plurality of trans wells; an electrolyte solution including a redox-inactive buffer that includes anions having a diameter greater than a diameter of a constriction zone of the nanopore and a redox species. It is to be understood that any features of the any of the examples set forth herein may be combined together in any desirable manner. For example, any combination of features of the first example and/or of the second example and/or of the third example and/or of the fourth example may be used together, and/or may be combined with any of the other examples disclosed herein to achieve the benefits as described in this disclosure, including, for example, controlling the depletion of an electrolyte species. BRIEF DESCRIPTION OF THE DRAWINGS Features of examples of the present disclosure will become apparent by reference to the following detailed description and drawings, in which like reference numerals correspond to similar, though perhaps not identical, compo