# US 9,434,990 B2 · Apparatus and method for molecular separation, purification, and sensing

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- Number: US 9,434,990 B2
- Document ID: US9434990B2
- Status: Granted
- Kind: B2
- Application number: 13/437,817
- Filed: 2012-04-02
- Published: 2013-10-03
- Granted: 2016-09-06
- Earliest claimed: 2012-04-02
- Assignees: Lux Bio Group, Inc.
- Applicants: Gordon Holt
- Inventors: Gordon Holt
- Web page: https://patentgrep.com/patent/US9434990B2

## CPC

- G01N 33/48721: {Investigating individual macromolecules, e.g. by translocation through nanopores (Coulter counters in general G01N15/12; fabrication methods for nanoscale apertures B81B1/00; sequencing of nucleic acids C12Q1/68)}
- B82Y 15/00: Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
- B82Y 40/00: Manufacture or treatment of nanostructures
- C12Q 1/68: involving nucleic acids
- C12Q 1/6869: Methods for sequencing
- C12Q 1/6874: involving nucleic acid arrays, e.g. sequencing by hybridisation
- G01N 27/44791: {Microapparatus (sample containers with integrated microfluidic structures B01L3/5027)}
- B82Y 5/00: Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
- B82Y 30/00: Nanotechnology for materials or surface science, e.g. nanocomposites
- G01N 27/3271: {Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood (amperometry per se G01N27/49; aspects concerning the enzyme reagent C12Q1/001)}
- G01N 27/403: Cells and electrode assemblies
- G01N 33/5438: {Electrodes}
- Y10T 436/143333: Saccharide [e.g., DNA, etc.]
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- B82Y: SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

Described are devices and methods for forming one or more nanomembranes including electroactive nanomembranes within a nanowell or nanotube, or combinations thereof, in a support material. Nanopores/nanochannels can be formed by the electroactive nanomembrane within corresponding nanowells. The electroactive nanomembrane is capable of controllably altering a dimension, a composition, and/or a variety of properties in response to electrical stimuli. Various embodiments also include devices/systems and methods for using the nanomembrane-containing devices for molecular separation, purification, sensing, etc.

## Claims

### Claim 1

A method comprising:
providing a nanochannel within a support material, wherein the nanochannel comprises one or more sidewall electrodes;
disposing a nanomembrane inside the nanochannel, wherein the nanomembrane encircles a nanopore, has a hydrophobic surface, and is in direct contact with at least a portion of one sidewall electrode of the nanochannel;
tuning a size of the nanopore by tuning dimensionality of the nanomembrane with an electrical stimulus on the nanomembrane;
tethering a lipid bilayer on the hydrophobic surface, the lipid bilayer spanning the nanopore;
disposing a protein nanopore into the lipid bilayer;
wherein the size of the nanopore is tuned such that the lipid bilayer spanning the nanopore only accommodates a single protein nanopore.

### Claim 2 (depends on 1)

The method of claim 1, further comprising stabilizing the lipid bilayer.

### Claim 3 (depends on 1)

The method of claim 1, wherein the protein nanopore comprises an α-hemolysin or a mycobacterium smegmatis porin A.

### Claim 4 (depends on 1)

The method of claim 1, further comprising disrupting the lipid bilayer by attracting, repulsing, or electrophoresing charged molecules to or from the lipid bilayer, by an electromotive force on the lipid bilayer or by a temperature change.

### Claim 5 (depends on 1)

The method of claim 1, wherein forming the nanomembrane is by a method selected from a group consisting of spraying, vapor-phase deposition, sputtering, spin coating, precipitation, in situ polymerization, and a combination thereof.

### Claim 6 (depends on 1)

The method of claim 1, wherein a mechanical, a chemical or electrical properties of the nanomembrane is uniform.

### Claim 7 (depends on 1)

The method of claim 1, wherein the nanochannel further comprises one or more bottom electrodes exposed to the nanochannel.

### Claim 8 (depends on 1)

The method of claim 1, wherein the nanomembrane comprises two layers of distinct properties.

### Claim 9 (depends on 1)

The method of claim 1, wherein the electrical stimulus comprises an electrical current, a voltage, or an electrical waveform.

### Claim 10 (depends on 1)

The method of claim 1, wherein at least a portion of the nanomembrane is electrically conductive.