# US 10,767,160 B2 · Methods for modulating production profiles of recombinant proteins

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- Number: US 10,767,160 B2
- Document ID: US10767160B2
- Status: Granted
- Kind: B2
- Application number: 15/743,714
- Filed: 2016-07-07
- Published: 2018-07-19
- Granted: 2020-09-08
- Earliest claimed: 2015-07-17
- Assignees: Ares Trading S.A.
- Applicants: Ares Trading S.A.
- Inventors: Anais Muhr; David Bruhlmann; Martin Jordan; Herve Broly; Matthieu Stettler
- Web page: https://patentgrep.com/patent/US10767160B2

## CPC

- C12N 5/0018: {Culture media for cell or tissue culture (media for specific animal cell type C12N5/06)}
- C07K 16/18: against material from animals or humans
- C07K 16/28: against receptors, cell surface antigens or cell surface determinants
- C12N 15/67: General methods for enhancing the expression
- C07K 2317/21: from primates, e.g. man
- C07K 2317/24: containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
- C12N 2501/999: Small molecules not provided for elsewhere
- C12N 2511/00: Cells for large scale production
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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

The invention is in the field of cell culture. Particularly the invention relates to methods of culturing a host cell expressing a recombinant protein in a cell culture medium comprising an effective amount of a triazine dye or supplemented with an effective amount of a triazine dye, whereby production of said protein is increased relative to cells grown without said triazine dye or any other inducer.

## Claims

### Claim 1

A method of increasing production of a recombinant protein, said method comprising culturing a mammalian host cell expressing said protein in a cell culture medium comprising a triazine dye, wherein the triazine dye is selected from the group consisting of Cibacron Blue 3GA and Reactive Red 120 and wherein the triazine dye is present at a concentration of 0.01 μM to 150 μM, wherein production of the recombinant protein is thereby increased.

### Claim 2 (depends on 1)

The method of claim 1, wherein the host cell is Chinese Hamster Ovary (CHO) cells.

### Claim 3 (depends on 1)

The method of claim 1, wherein the recombinant protein is selected from the group consisting of an antibody or antigen binding fragment thereof, a human antibody or antigen-binding portion thereof, a humanized antibody or antigen-binding portion thereof, a chimeric antibody or antigen-binding portion thereof, a recombinant fusion protein, a growth factor, a hormone, and a cytokine.