Low acidic species compositions and methods for producing and using the same

Patent metadata

Field	Value
Publication number	US-9315574-B2
Application number	US-201414584619-A
Country	US
Kind code	B2
Filing date	Dec 29, 2014
Priority date	Oct 18, 2013
Publication date	Apr 19, 2016
Grant date	Apr 19, 2016

Abstract

The instant invention relates to low acidic species (AR) compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, and methods, e.g., cell culture and/or protein purification methods, for producing such low AR compositions. Methods for using such compositions to treat a disorder, e.g., a disorder in which TNFα is detrimental, are also provided.

First claim

The invention claimed is:

1. A method for producing a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, the method comprising:
contacting a first sample comprising the immunoglobulin, wherein the sample comprises more than 10% total acidic species of the immunoglobulin, to a first chromatography media in the presence of a loading buffer to produce a first chromatography sample,
wherein the first chromatography media is selected from the group consisting of an ion exchange chromatography media, an affinity chromatography media and a hydrophobic interaction chromatography (HIC) media,
wherein the first chromatography sample comprises a composition of the immunoglobulin comprising less than 10% total acidic species of the immunoglobulin, wherein the acidic species of the immunoglobulin correspond to the peaks that elute earlier than the main peak in a WCX-10 HPLC chromatogram of the immunoglobulin, 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.

2. The method of claim 1 , wherein producing the first chromatography sample comprises washing the first chromatography media with a wash buffer.

3. The method of claim 2 , wherein the wash buffer is the same as the loading buffer.

4. The method of claim 1 , wherein the first chromatography sample is a flow through chromatography sample which comprises less than 10% total acidic species of the immunoglobulin.

5. The method of claim 1 , wherein producing the first chromatography sample comprises eluting the first chromatography sample, thereby producing a first eluted sample which comprises less than 10% total acidic species of the immunoglobulin.

6. The method of claim 5 , further comprising contacting the first eluted sample to a second chromatography media and eluting the sample, thereby producing a second eluted sample, wherein the second chromatography media is selected from the group consisting of an ion exchange chromatography media, an affinity chromatography media and a hydrophobic interaction chromatography (HIC) media.

7. The method of claim 6 , further comprising contacting the second eluted sample to a third chromatography media and eluting the sample, thereby producing a third eluted sample, wherein the third chromatography media is selected from the group consisting of an ion exchange chromatography media, an affinity chromatography media and a hydrophobic interaction chromatography (HIC) media.

8. The method of claim 7 , wherein the first chromatography media is an affinity chromatography media, the second chromatography media is an ion exchange chromatography media and the third chromatography media is a hydrophobic interaction chromatography (HIC) media.

9. The method of claim 7 , wherein the first chromatography media is an affinity chromatography media, the second chromatography media is an ion exchange chromatography media and the third chromatography media is an ion exchange chromatography media.

10. The method of claim 9 , wherein the affinity chromatography media is a protein A chromatography media, the ion exchange chromatography media used as the second chromatography media is an anion exchange (AEX) chromatography media and the ion exchange chromatography media used as the third chromatography media is a cation exchange (CEX) chromatography media.

11. The method of claim 1 , wherein the first chromatography media is an ion exchange chromatography media selected from the group consisting of an anion exchange (AEX) chromatography adsorbent material, a cation exchange (CEX) chromatography adsorbent material, a cation exchange mixed mode media, and an anion exchange mixed mode media.

12. The method of claim 11 , wherein the cation exchange (CEX) adsorbent material is selected from the group consisting of a CEX resin and a CEX membrane adsorber.

13. The method of claim 11 , wherein the anion exchange (AEX) adsorbent material is selected from the group consisting of an AEX resin and an AEX membrane adsorber.

14. The method of claim 1 , wherein the first chromatography media is an affinity chromatography media.

15. The method of claim 14 , wherein the affinity chromatography media is a Protein A chromatography media.

16. The method of claim 1 , wherein the first chromatography media is a hydrophobic interaction chromatography (HIC) media.

17. The method of claim 1 , wherein the first chromatography media is a mixed mode media comprising ion exchange and hydrophobic interaction functional groups.

18. The method of claim 1 , wherein the first chromatography media is a CEX adsorbent material or a mixed mode media, and the pH of the loading buffer is lower than the isoelectric point of adalimumab.

19. The method of claim 1 , wherein the immunoglobulin is adalimumab and wherein the total acidic species of adalimumab comprise a first acidic region (AR1) and a second acidic region (AR2).

20. The composition of claim 19 , wherein the first acidic region (AR1) and the second acidic region (AR2) comprise charge variants, structure variants and fragmentation variants.

21. The composition of claim 20 , wherein the charge variants comprise one or more of deamidation variants, glycation variants, afucosylation variants, MGO variants or citric acid variants, the structure variants comprise one or more of glycosylation variants or acetonation variants, and the fragmentation variants comprise one or more of Fab fragment variants, C-terminal truncation variants or variants missing a heavy chain variable domain.

22. The method of claim 1 , wherein the composition comprises 9% or less, 8% or less, 7% or less, 6% or less, 5% or less, 4.5% or less, 4% or less, 3.5% or less, 3% or less, 2.5% or less, or 2% or less total acidic species of the immunoglobulin comprising the 6 CDR domains of adalimumab.

23. The method of claim 1 , wherein the immunoglobulin is adalimumab.

24. The method of claim 1 , wherein the composition comprises 1.4% to 10% total acidic species of the immunoglobulin comprising the 6 CDR domains of adalimumab.

25. The method of claim 1 , wherein the composition comprises 1.4% or less total acidic species of the immunoglobulin comprising the 6 CDR domains of adalimumab.

26. A method for producing a composition comprising an immunoglobulin comprising the 6 CDR domains of adalimumab, the method comprising:
(a) contacting a first sample comprising the immunoglobulin, wherein the sample comprises more than 10% total acidic species of the immunoglobulin, to an affinity chromatography media in a first loading buffer, and eluting a first eluted sample from the affinity chromatography media;
(b) contacting the first eluted sample to a second chromatography media in a second loading buffer and eluting a second eluted sample from the second chromatography media; and
(c) contacting the second eluted sample to a third chromatography media in a third loading buffer and eluting a third eluted sample from the third chromatography media;
wherein the third eluted sample comprises a composition of the immunoglobulin comprising less than 10% total acidic species of the immunoglobulin, wherein the acidic species of the immunoglobulin correspond to the peaks that elute earlier than the main peak in a W

Assignees

• Abbvie Inc

Inventors

• Ramasubramanyan Natarajan
• Yang Lihua
• Herigstad Matthew Omon
• Yang Hong
• Subramanian Kartik
• Zeng Xiaobei
• Dong Diane D
• Lim Wen Chung
• Gifford Kathreen A
• Kaymakcalan Zehra
• Chumsae Christopher

Classifications

• C07K16/241 Primary

  Tumor Necrosis Factors · CPC title

• A61K47/26

  Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin · CPC title

• C07K2317/10

  characterized by their source of isolation or production · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title