# US 11,111,269 B2 · Hydrophobic interaction protein chromatography under no-salt conditions

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- Number: US 11,111,269 B2
- Document ID: US11111269B2
- Status: Granted
- Kind: B2
- Application number: 16/005,642
- Filed: 2018-06-11
- Published: 2019-05-16
- Granted: 2021-09-07
- Earliest claimed: 2013-03-15
- Assignees: Biogen MA Inc.
- Applicants: Biogen MA Inc.
- Inventors: Sanchayita Ghose; Yinying Tao
- Web page: https://patentgrep.com/patent/US11111269B2

## CPC

- C07K 1/20: Partition-, reverse-phase or hydrophobic interaction chromatography
- C07K 16/065: {Purification, fragmentation}
- 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

Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

## Claims

### Claim 1

A method of purifying an antibody, the method comprising:
subjecting an antibody in solution to hydrophobic interaction chromatography in flow through mode using a matrix containing hydrophobic ligands with a mobile phase buffer, wherein the mobile phase buffer does not contain a kosmotropic salt and has a pH of about 5.0 to about 7.0.

### Claim 2

A method of purifying an antibody, the method comprising:
providing an antibody in a solution;
loading the solution onto a matrix containing hydrophobic ligands;
adding a mobile phase buffer that has a pH of about 5.0 to about 7.0, wherein the mobile phase buffer does not contain a kosmotropic salt; and
collecting a flow through fraction that contains the antibody.

### Claim 3 (depends on 1, 2)

The method of claim 1 or 2, wherein the pH of the mobile phase buffer is about 5.0 to about 6.5, about 5.0 to about 6.0, about 5.5 to about 7.0, or about 6.0 to about 7.0.

### Claim 4 (depends on 1)

The method of claim 1, wherein the matrix comprises a hydroxylated methacrylic polymer, agarose or sepharose.

### Claim 5 (depends on 1)

The method of claim 1, wherein the hydrophobic ligands are ether groups, polypropylene glycol groups, phenyl groups, butyl groups, hexyl groups or octyl groups.

### Claim 6 (depends on 1)

The method of claim 1, wherein the antibody is a monoclonal antibody or a polyclonal antibody.

### Claim 7 (depends on 6)

The method of claim 6, wherein the antibody is a human antibody, a mouse antibody or a chimeric antibody.

### Claim 8 (depends on 1)

The method of claim 1 further comprising collecting a flow through fraction that contains the antibody.

### Claim 9 (depends on 1)

The method of claim 1, wherein the hydrophobic ligands comprise straight chain alkyl ligands or aryl ligands.

### Claim 10 (depends on 1)

The method of claim 1, wherein the matrix is a polymeric matrix.

### Claim 11 (depends on 1)

The method of claim 1, wherein the kosmotropic salt is selected from the group consisting of ammonium sulfate, sodium citrate, and potassium phosphate.

### Claim 12 (depends on 2)

The method of claim 2, wherein the kosmotropic salt is selected from the group consisting of ammonium sulfate, sodium citrate, and potassium phosphate.