Compositions comprising purified recombinant IL-13 antibody

Patent metadata

Field	Value
Publication number	US-10597447-B2
Application number	US-201815902139-A
Country	US
Kind code	B2
Filing date	Feb 22, 2018
Priority date	Sep 13, 2013
Publication date	Mar 24, 2020
Grant date	Mar 24, 2020

Abstract

Purified recombinant polypeptides isolated from Chinese hamster ovary host cells, including antibodies, such as therapeutic antibodies, and methods of making and using such polypeptides are provided.

First claim

What is claimed is:

1. A composition comprising an anti-IL13 monoclonal antibody purified from Chinese hamster ovary host cells, wherein the composition comprises the anti-IL13 antibody and a residual amount of hamster phospholipase B-like 2 (PLBL2), wherein the residual amount of hamster PLBL2 is less than 20 ng/mg, wherein the anti-IL13 antibody comprises three heavy chain CDRs, CDR-H1 having the amino acid sequence of SEQ ID NO.: 1, CDR-H2 having the amino acid sequence of SEQ ID NO.: 2, and CDR-H3 having the amino acid sequence of SEQ ID NO.: 3, and three light chain CDRs, CDR-L1 having the amino acid sequence of SEQ ID NO.: 4, CDR-L2 having the amino acid sequence of SEQ ID NO.: 5, and CDR-L3 having the amino acid sequence of SEQ ID NO.: 6.

2. The composition of claim 1 , wherein the anti-IL13 antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO.: 7.

3. The composition of claim 1 , wherein the anti-IL13 antibody comprises a light chain variable region having the amino acid sequence of SEQ ID NO.: 9.

4. The composition of claim 2 , wherein the anti-IL13 antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO.: 10.

5. The composition of claim 3 , wherein the anti-IL13 antibody comprises a light chain having the amino acid sequence of SEQ ID NO.: 14.

6. The composition of claim 1 , wherein the anti-IL13 antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO.: 7 and a light chain variable region having the amino acid sequence of SEQ ID NO.: 9.

7. The composition of claim 6 , wherein the anti-IL13 antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO.: 10 and a light chain having the amino acid sequence of SEQ ID NO.: 14.

8. The composition of claim 1 , wherein the amount of hamster PLBL2 was quantified using an immunoassay or a mass spectrometry assay.

9. The composition of claim 8 , wherein the immunoassay is a total Chinese hamster ovary protein ELISA or a hamster PLBL2 ELISA.

10. The composition of claim 8 , wherein the mass spectrometry assay is LC-MS/MS.

11. A purified anti-IL13 monoclonal antibody preparation isolated from Chinese hamster ovary host cells, wherein the preparation is purified by a process comprising a hydrophobic interaction chromatography (HIC) step thereby producing a purified preparation, wherein the purified preparation comprises the anti-IL13 antibody and a residual amount of hamster PLBL2, wherein the residual amount of hamster PLBL2 is less than 20 ng/mg, wherein the anti-IL13 antibody comprises three heavy chain CDRs, CDR-H1 having the amino acid sequence of SEQ ID NO.: 1, CDR-H2 having the amino acid sequence of SEQ ID NO.: 2, and CDR-H3 having the amino acid sequence of SEQ ID NO.: 3, and three light chain CDRs, CDR-L1 having the amino acid sequence of SEQ ID NO.: 4, CDR-L2 having the amino acid sequence of SEQ ID NO.: 5, and CDR-L3 having the amino acid sequence of SEQ ID NO.: 6.

12. The anti-IL13 antibody preparation of claim 11 , wherein the HIC step comprises PHENYL SEPHAROSE™ 6 Fast Flow (High Sub) resin.

13. The anti-IL13 antibody preparation of claim 12 , wherein the HIC step comprises operating a PHENYL SEPHAROSE™ 6 Fast Flow (High Sub) resin-containing column in flow-through mode.

14. The anti-IL13 antibody preparation of claim 13 , wherein the HIC step comprises an equilibration buffer and a wash buffer, wherein each of the equilibration buffer and the wash buffer comprise 50 mM sodium acetate pH 5.0.

15. The anti-IL13 antibody preparation of claim 14 , wherein the flow-through is monitored by absorbance at 280 nanometers and the flow-through is collected between 0.5 OD to 1.5 OD.

16. The anti-IL13 antibody preparation of claim 14 , wherein the flow-through is collected for a maximum of 8 column volumes.

17. The anti-IL13 antibody preparation of claim 11 , wherein the process further comprises an affinity chromatography step and/or an ion exchange chromatography step.

18. The anti-IL13 antibody preparation of claim 17 , wherein the affinity chromatography is protein A chromatography.

19. The anti-IL13 antibody preparation of claim 17 , wherein the ion exchange chromatography is anion exchange chromatography.

20. A purified anti-IL13 monoclonal antibody preparation isolated from Chinese hamster ovary cells, wherein the antibody preparation is purified by a process comprising a first Protein A affinity chromatography step, a second anion exchange chromatography step, and a third hydrophobic interaction chromatography (HIC) step thereby producing a purified preparation, wherein the purified preparation comprises the anti-IL13 antibody and a residual amount of hamster PLBL2, wherein the amount of hamster PLBL2 is less than 20 ng/mg, wherein the anti-IL13 antibody comprises three heavy chain CDRs, CDR-H1 having the amino acid sequence of SEQ ID NO.: 1, CDR-H2 having the amino acid sequence of SEQ ID NO.: 2, and CDR-H3 having the amino acid sequence of SEQ ID NO.: 3, and three light chain CDRs, CDR-L1 having the amino acid sequence of SEQ ID NO.: 4, CDR-L2 having the amino acid sequence of SEQ ID NO.: 5, and CDR-L3 having the amino acid sequence of SEQ ID NO.: 6.

21. The anti-IL13 antibody preparation of claim 20 , wherein the affinity chromatography step comprises MABSELECT SURE™ resin, the anion exchange chromatography step comprises Q SEPHAROSE™ Fast Flow resin, and the HIC step comprises PHENYL SEPHAROSE™ 6 Fast Flow (high sub) resin.

22. The anti-IL13 antibody preparation of claim 21 , wherein:
the affinity chromatography step comprises operating a MABSELECT SURE™ resin-containing column in bind-elute mode;
the anion exchange chromatography step comprises operating a Q SEPHAROSE™ Fast Flow resin-containing column in bind-elute mode; and
the HIC step comprises operating a PHENYL SEPHAROSE™ 6 Fast Flow (High Sub) resin-containing column in flow-through mode.

23. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 15 ng/mg.

24. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 10 ng/mg.

25. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 8 ng/mg.

26. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 5 ng/mg.

27. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 3 ng/mg.

28. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 2 ng/mg.

29. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 1 ng/mg.

30. The composition of claim 1 , wherein the residual amount of hamster PLBL2 is less than 0.5 ng/mg.

31. The purified anti-IL13 monoclonal antibody preparation of claim 11 , wherein the anti-IL13 antibody comprises a heavy chain variable region having the amino acid sequence of SEQ ID NO.: 7 and a light chain variable region having the amino acid sequence of SEQ ID NO.: 9.

32. The purified anti-IL13 monoclonal antibody preparation of claim 11 , wherein the anti-IL13 antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO.: 10 and a light chain having the amino acid sequence of SEQ ID NO.: 14.

33. The purified anti-IL13 monoclonal antibody preparatio

Assignees

• Genentech Inc

Inventors

• Yu X Christopher
• Fisher Susan C
• Sanchez Ailen M
• Vanderlaan Martin

Classifications

• A61P37/08

  Antiallergic agents (antiasthmatic agents A61P11/06; ophthalmic antiallergics A61P27/14) · CPC title

• A61P37/06

  Immunosuppressants, e.g. drugs for graft rejection · CPC title

• C07K1/18 Primary

  Ion-exchange chromatography · CPC title

• A61P31/04

  Antibacterial agents · CPC title

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title