# US 2019/0169686 A1 · Hybrid nanopores and uses thereof for detection of analytes

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- Number: US 2019/0169686 A1
- Document ID: US20190169686A1
- Status: Published application
- Kind: A1
- Application number: 16/214,710
- Filed: 2018-12-10
- Published: 2019-06-06
- Earliest claimed: 2013-02-07
- Assignees: Yissum Research Development Company of the Hebrew University of Jerusalem
- Applicants: Yissum Research Development Company of the Hebrew University of Jerusalem Ltd.
- Inventors: Danny PORATH; Oded SHOSEYOV; Yuval NEVO; Ke LIU; Dvir MAROM ROTEM
- Web page: https://patentgrep.com/patent/US20190169686A1

## CPC

- C12Q 1/6869: Methods for sequencing
- G01N 27/44704: {Details; Accessories}
- G01N 27/44756: {Apparatus specially adapted therefor}
- 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)}
- 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
- 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)

## Abstract

The invention relates to a hybrid structure comprising perforated solid substrate having at least one nanopore perforating therethrough, and devices and uses thereof.

## Claims

### Claim 1

A method for analysis of at least one analyte in a sample comprising:
(i) applying a sample comprising at least one analyte or suspected to comprise at least one analyte onto a hybrid structure, wherein the hybrid structure comprises (a) a solid substrate having at least one nanopore perforating therethrough, and (b) at least one ring-like polypeptide situated at a region of said at least one nanopore, said region being selected from an opening of the nanopore and an interior region of said nanopore, wherein the binding of the at least one ring-like polypeptide to the nanopore is different from covalent binding,
(ii) permitting the sample to flow through the nanopore;
(iii) determining at least one of (a) presence or absence of an analyte in the sample, (b) identity of the analyte in the sample and (c) concentration of the analyte in the sample.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the sample comprising at least one ring-like polypeptide and the hybrid structure is formed in parallel with step (i).

### Claim 3 (depends on 1)

The method according to claim 1, comprising monitoring at least one measurable parameter related to the nanopore.

### Claim 4 (depends on 3)

The method according to claim 3, wherein the at least one measurable parameter related to the nanopore is used to obtain at least one data value.

### Claim 5 (depends on 3)

The method according to claim 3, wherein the at least one measurable parameter is at least one of (i) change in current, (ii) time duration of change in the current and any combination thereof.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the ring-like polypeptide is stable protein 1 (SP1).

### Claim 7 (depends on 1)

The method according to claim 1, wherein the analyte is a protein, a polypeptide, an oligopeptide, a peptide, a ganglioside, a lipid, a phospholipid, a carbohydrate, a small molecule or a nucleic acid.

### Claim 8 (depends on 1)

The method according to claim 1, wherein the analyte is a double stranded nucleic acid or a single stranded nucleic acid.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the analyte is DNA or RNA.

### Claim 10 (depends on 1)

The method according to claim 1, wherein the analyte is nucleic acid and is in a linear conformation.

### Claim 11 (depends on 1)

The method according to claim 1, wherein the sample is selected from bodily fluid, an isolated cell population and a cell preparation.

### Claim 12 (depends on 1)

The method according to claim 1, wherein the sample is bodily fluid and is selected from the group consisting of blood, serum, plasma, urine, cerebrospinal fluid, amniotic fluid, tear fluid, nasal wash, mucus, saliva, sputum, broncheoalveolar fluid, throat wash, vaginal fluid and semen.

### Claim at position 13

13-19. (canceled)