# US 2010/0168403 A1 · Protein refolding agent and refolding method

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2010/0168403 A1
- Document ID: US20100168403A1
- Status: Published application
- Kind: A1
- Application number: 12/659,247
- Filed: 2010-03-02
- Published: 2010-07-01
- Earliest claimed: 2008-03-19
- Inventors: Shunichlro Yamaguchi; Shuii Mjura
- Web page: https://patentgrep.com/patent/US20100168403A1

## CPC

- C07K 1/1136: {by reversible modification of the secondary, tertiary or quarternary structure, e.g. using denaturating or stabilising agents}
- 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

A refolding agent and refolding method which makes it possible to produce high-purity proteins in high productivity. The refolding agent includes a phosphorus-containing compound (A) and an oxycarbonyl group-containing compound (B). The refolding method includes the step of treating the unfolded protein with the refolding agent. As the compound (A), there may be at least one selected from inorganic phosphoric acids, alkyl phosphate esters, sugar phosphate esters, and salts of these, and as the compound (B), there may be at least one selected from formic acid, acetic acid, propionic acid, lactic acid, tartaric acid, and salts of these.

## Claims

### Claim 1

A refolding agent for an unfolded protein which is a phosphorus-containing refolding agent (C) comprising a compound (A) having a group represented by the formula (1):

### Claim 2 (depends on 1)

The refolding agent according to claim 1 wherein, in a refolding step, the concentration thereof at a compound (A) concentration, in the system, is 0.2 to 6 moles/L.

### Claim 3 (depends on 1)

The refolding agent according to claim 1
wherein the compound (A) is at least one species selected from the group consisting of inorganic phosphoric acids, alkyl phosphate esters, sugar phosphate esters, and salts of these.

### Claim 4 (depends on 1, 2)

The refolding agent according to claims 1 or 2
wherein the compound (A) is at least one species selected from the group consisting of phosphoric acid, pyrophosphoric acid, polyphosphoric acids, adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, and salts of these.

### Claim at position 5

5-8. (canceled)

### Claim 9 (depends on 1, 2)

The refolding agent according to claims 1 or 2
wherein the unfolded protein has a molecular weight of 1,000 to 300,000.

### Claim 10 (depends on 1, 2)

The refolding agent according to claims 1 or 2
wherein the unfolded protein is a protein unfolded with guanidine hydrochloride and/or urea.

### Claim 11 (depends on 1, 2)

A method of refolding an unfolded protein
which comprises the step of treating the unfolded protein with the phosphorus-containing refolding agent (C) according to claims 1 or 2,
wherein, in this step, the concentration thereof at a compound (A) concentration, in the system, is 0.2 to 6 moles/L.

### Claim 12

(canceled)

### Claim 13 (depends on 11)

The method according to claim 11
wherein a surfactant (E) is used in the step of treating with the refolding agent.

### Claim 14 (depends on 11)

The method according to claim 11
wherein a pH adjusting agent (F) and/or protein stabilizing agent (G) is used in the step of treating with the refolding agent.

### Claim 15 (depends on 11)

The method according to claim 11
wherein, in the step of treating with the phosphorus-containing refolding agent (C) or the oxycarbonyl group-containing refolding agent (D), the protein concentration in the system is 0.2 to 30 mg/mL.

### Claim 16 (depends on 11)

A method of protein production
which comprises the step of refolding by the method according to claim 11.

### Claim 17 (depends on 11)

A protein obtained by the production method according to claim 11.