# US 2010/0167346 A1 · Method of producing heterogeneous protein

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- Number: US 2010/0167346 A1
- Document ID: US20100167346A1
- Status: Published application
- Kind: A1
- Application number: 12/733,052
- Filed: 2008-08-06
- Published: 2010-07-01
- Earliest claimed: 2007-08-07
- Assignees: Chugai Seiyaku Kabushiki Kaisha
- Inventors: Hisahiro Tabuchi; Tomoya Sugiyama
- Web page: https://patentgrep.com/patent/US20100167346A1

## CPC

- C07K 16/28: against receptors, cell surface antigens or cell surface determinants
- C07K 14/47: from mammals
- C12N 9/1096: {transferring nitrogenous groups (2.6)}
- C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione
- C12Y 206/01002: Alanine transaminase (2.6.1.2), i.e. alanine-aminotransferase
- Y02P 20/52: using catalysts, e.g. selective catalysts
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12Y: ENZYMES
- Y02P: CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS

## Abstract

The present invention provides a method capable of producing a natural or recombinant protein in high yield. The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses alanine aminotransferase and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide.

## Claims

### Claim 1

A method of producing a desired polypeptide, comprising culturing a cell which strongly expresses alanine aminotransferase and has a transferred DNA encoding the desired polypeptide and thereby allowing the cell to produce said desired polypeptide.

### Claim 2 (depends on 1)

The method of claim 1, wherein the cell which strongly expresses alanine aminotransferase is a cell into which a DNA encoding the alanine aminotransferase has been transferred.

### Claim 3 (depends on 1)

The method of claim 1, wherein the cells that strongly express alanine aminotransferase further express a taurine transporter strongly.

### Claim 4 (depends on 3)

The method of claim 3, wherein the cells that strongly express a taurine transporter are cells into which DNA encoding a taurine transporter has been transferred.

### Claim 5 (depends on 2)

The method of claim 2, wherein the cell is Chinese hamster ovary cells.

### Claim 6 (depends on 1)

The method of claim 1, wherein the desired polypeptide is an antibody.

### Claim 7 (depends on 1)

The method of claim 1, wherein the DNA encoding the alanine aminotransferase is any one of the following (a) to (e):
(a) a DNA encoding a polypeptide having the amino acid sequence as shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58 or 60;
(b) a DNA encoding a polypeptide which has an amino acid sequence derived from the amino acid sequence as shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58 or 60 by substitution, deletion, addition and/or insertion of one or more amino acid residues and yet has alanine aminotransferase activity;
(c) a DNA encoding a polypeptide having 70% or more amino acid sequence homology with the amino acid sequence as shown in SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58 or 60 and yet having alanine aminotransferase activity;
(d) a DNA having the nucleotide sequence as shown in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57 or 59;
(e) a DNA which hybridizes to a DNA complementary to a DNA having the nucleotide sequence as shown in SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57 or 59 under stringent conditions and yet encodes a polypeptide having alanine aminotransferase activity.

### Claim 8 (depends on 1)

A method of preparing a pharmaceutical composition containing a polypeptide prepared by the method of claim 1.

### Claim 9

A cell which has a transferred DNA encoding alanine aminotransferase and a transferred DNA encoding a desired polypeptide.

### Claim 10 (depends on 9)

The cell according to claim 9, which further has a transferred DNA encoding a taurine transporter.

### Claim 11

A cell which has a transferred DNA encoding alanine aminotransferase and a transferred DNA encoding a taurine transporter.

### Claim 12

A method of producing a desired polypeptide, comprising culturing in an α-ketoglutarate-containing medium a cell which strongly expresses alanine aminotransferase and has a transferred DNA encoding the desired polypeptide and thereby allowing the cell to produce said desired polypeptide.