Cell culture methods to reduce acidic species

Patent metadata

Field	Value
Publication number	US-2016145331-A1
Application number	US-201615008895-A
Country	US
Kind code	A1
Filing date	Jan 28, 2016
Priority date	Apr 20, 2012
Publication date	May 26, 2016
Grant date	—

Abstract

The instant invention relates to the field of protein production and purification, and in particular to compositions and processes for controlling the amount of acidic species expressed by host cells, as well as to compositions and processes for controlling the amount of acidic species present in purified preparations.

First claim

What is claimed is:

1 . A composition comprising adalimumab, wherein the composition comprises less than 10% total acidic species of adalimumab,
wherein the acidic species of adalimumab do not include process-related impurities selected from the group consisting of host cells and lysed host cells, and wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab.

2 . The composition of claim 1 , wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.

3 . The composition of claim 1 , wherein the acidic species of adalimumab comprise a first acidic region (AR1) and a second acidic region (AR2).

4 . The composition of claim 1 , wherein the composition comprises 9% or less acidic species of adalimumab.

5 . The composition of claim 1 , wherein the composition comprises 6%-8% acidic species of adalimumab.

6 . The composition of claim 1 , wherein the adalimumab is produced in a mammalian host cell grown in cell culture.

7 . The composition of claim 6 , wherein the mammalian host cell is selected from the group consisting of a CHO cell, an NSO cell, a COS cell, and an SP2 cell.

8 . The composition of claim 7 , wherein the mammalian host cell is a CHO cell.

9 . The composition of claim 8 , wherein the composition is lyophilized.

10 . A pharmaceutical composition suitable for administration to a subject comprising the composition of claim 8 and a pharmaceutically acceptable carrier.

11 . The pharmaceutical composition of claim 10 , wherein adalimumab is present in the pharmaceutical composition at a concentration of 0.1-250 mg/ml.

12 . The pharmaceutical composition of claim 10 , wherein the pharmaceutical composition comprises one or more excipient.

13 . The pharmaceutical composition of claim 12 , wherein the one or more excipient is selected from the group consisting of a buffer, an isotonic agent, a surfactant or a combination thereof.

14 . The pharmaceutical composition of claim 13 , wherein the pharmaceutical composition comprises the surfactant polysorbate 80.

15 . The pharmaceutical composition of claim 13 , wherein the pharmaceutical composition comprises an amino acid buffer.

16 . The pharmaceutical composition of claim 15 , wherein the amino acid is histidine.

17 . The pharmaceutical composition of claim 13 , wherein the pharmaceutical composition comprises the isotonic agent mannitol.

18 . A method for treating a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 10 , thereby treating the subject.

19 . A composition comprising adalimumab, wherein the composition comprises less than 10% total acidic species of adalimumab,
wherein the acidic species of adalimumab do not include process-related impurities selected from the group consisting of host cells and lysed host cells, wherein the acidic species of adalimumab correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of adalimumab, and wherein the WCX-10 HPLC chromatogram is generated using a first mobile phase of 10 mM Sodium Phosphate dibasic (pH 7.5) and a second mobile phase of 10 mM Sodium Phosphate dibasic, 500 mM Sodium Chloride (pH 5.5), and wherein the WCX-10 HPLC chromatogram is generated using detection at 280 nm.

20 . The composition of claim 19 , wherein the acidic species of adalimumab comprise a first acidic region (AR1) and a second acidic region (AR2).

21 . The composition of claim 19 , wherein the composition comprises 9% or less acidic species of adalimumab.

22 . The composition of claim 19 , wherein the composition comprises 6%-8% acidic species of adalimumab.

23 . The composition of claim 19 , wherein the adalimumab is produced in a mammalian host cell grown in cell culture.

24 . The composition of claim 23 , wherein the mammalian host cell is selected from the group consisting of a CHO cell, an NSO cell, a COS cell, and an SP2 cell.

25 . The composition of claim 24 , wherein the mammalian host cell is a CHO cell.

26 . A pharmaceutical composition suitable for administration to a subject comprising the composition of claim 25 and a pharmaceutically acceptable carrier.

27 . The pharmaceutical composition of claim 26 , wherein adalimumab is present in the pharmaceutical composition at a concentration of 0.1-250 mg/ml.

28 . The pharmaceutical composition of claim 26 , wherein the pharmaceutical composition comprises one or more excipient.

29 . The pharmaceutical composition of claim 28 , wherein the one or more excipient is selected from the group consisting of a buffer, an isotonic agent, a surfactant or a combination thereof.

30 . A method for treating a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 26 , thereby treating the subject.

Assignees

• Abbvie Inc

Inventors

• Subramanian Kartik
• Zeng Xiaobei
• Dong Diane D
• Lim Wen Chung
• Gifford Kathreen A
• Chumsae Christopher

Classifications

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K1/20

  Partition-, reverse-phase or hydrophobic interaction chromatography · CPC title

• C07K1/18

  Ion-exchange chromatography · CPC title

• C07K16/241 Primary

  Tumor Necrosis Factors · CPC title

• C12P21/00

  Preparation of peptides or proteins (single cell protein C12N1/00) · CPC title