# US 10,481,148 B2 · Cell line stably expressing Nav1.5 and method of screening therapeutic agent using the cell line

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- Number: US 10,481,148 B2
- Document ID: US10481148B2
- Status: Granted
- Kind: B2
- Application number: 15/444,335
- Filed: 2017-02-28
- Published: 2018-03-22
- Granted: 2019-11-19
- Earliest claimed: 2016-09-22
- Assignees: The Catholic University of Korea Industry-Academic Cooperation Foundation
- Applicants: The Catholic University of Korea Industry-Academic Cooperation Foundation
- Inventors: Jin-Sung Choi; Jeong Hee Choi
- Web page: https://patentgrep.com/patent/US10481148B2

## CPC

- G01N 33/5014: {for testing toxicity}
- C07K 14/705: Receptors; Cell surface antigens; Cell surface determinants {(tumour specific antigens C07K14/4748)}
- G01N 33/48728: {Investigating individual cells, e.g. by patch clamp, voltage clamp (investigating individual particles in general G01N15/10)}
- G01N 33/6872: {Intracellular protein regulatory factors and their receptors, e.g. including ion channels}
- G01N 2333/705: Assays involving receptors, cell surface antigens or cell surface determinants
- G01N 2500/04: Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
- G01N 2500/10: involving cells
- 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)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)

## Abstract

Provided are a cell line stably expressing voltage-gated sodium channel (Nav1.5), wherein a SCN5A gene (NCBI accession no. NM_198056) encoding Nav1.5 and a puromycin selection marker are inserted into a chromosome of a host cell, and a method of measuring cardiotoxicity of a test substance comprising: (1) contacting a test substance to the cell line of claim 1; and (2) measuring magnitude of voltage-gated sodium channel (Nav1.5) in the cell line in contact with the test substance.

## Claims

### Claim 1

A cell line stably expressing voltage-gated sodium channel (Nav1.5), the cell line having accession number KCTC 13095BP, wherein a SCN5A gene consisting of NCBI accession no. NM_198056 encoding Nav1.5 and a puromycin selection marker are inserted into a chromosome of the cell.

### Claim 2 (depends on 1)

The cell line of claim 1, wherein the cell is a human embryonic kidney 293 (HEK293) cell, and the cell line has sodium current magnitude in a range from about 5.5 nA to about 19.6 nA.

### Claim 3 (depends on 2)

The cell line of claim 2, wherein the cell line has average sodium current quantity of about 8.8±0.8 nA.

### Claim 4 (depends on 1)

A method of measuring cardiotoxicity of a test substance, the method comprising:
(1) contacting a test substance to the cell line of claim 1; and
(2) measuring the magnitude of the voltage-gated sodium channel Nav1.5 in the cell line in contact with the test substance.

### Claim 5 (depends on 1)

A method of screening an inhibitor for cardiotoxicity, the method comprising:
(1) contacting a test substance to the cell line of claim 1;
(2) measuring the magnitude of the voltage-gated sodium channel Nav1.5 in the cell line in contact with the test substance; and
(3) selecting the test substance having an increased magnitude of Nav1.5 current as compared to the magnitude of Nav1.5 current in a control sample.