Method for purifying active polypeptides or immunoconjugates

Patent metadata

Field	Value
Publication number	US-11136396-B2
Application number	US-202016786634-A
Country	US
Kind code	B2
Filing date	Feb 10, 2020
Priority date	Jul 30, 2010
Publication date	Oct 5, 2021
Grant date	Oct 5, 2021

Abstract

The present invention provides methods for isolating an active polypeptide or immunoconjugate by purification of a solution containing both the active polypeptide or immunoconjugate and an acidic variant thereof, such as a deamidated variant, using anion exchange chromatography. The present invention also provides compositions, formulations, and unit dosage forms comprising the purified polypeptide or immunoconjugate.

First claim

What is claimed is:

1. A method of treating a B cell malignancy comprising administering a composition comprising a purified polypeptide comprising an anti-CD22 antibody or antigen binding fragment thereof and a Pseudomonas exotoxin (PE) having an amino acid sequence selected from the group consisting of SEQ ID NOs: 16-22, wherein the composition comprises less than 25% deamidated species of the polypeptide.

2. The method of claim 1 , wherein the B cell malignancy is hairy cell leukemia.

3. The method of claim 1 , wherein the antibody or antigen binding fragment comprises a Fab, a Fab′, a F(ab′) 2 , a Fd, a single chain Fv or scFv, a disulfide linked Fv, a V-NAR domain, an IgNar, an intrabody, an IgGΔCH2, a minibody, a F(ab′) 3 , a tetrabody, a triabody, a diabody, a single-domain antibody, DVD-Ig, Fcab, mAb 2 , a (scFv) 2 , or a scFv-Fc.

4. The method of claim 1 , wherein the PE has the amino acid sequence of SEQ ID NO: 22.

5. The method of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a V H sequence and a V L sequence.

6. The method of claim 5 , wherein the V H sequence is selected from the group consisting of SEQ ID NOs: 6-11.

7. The method of claim 5 , wherein the V L sequence is selected from the group consisting of SEQ ID NOs: 2 and 12-15.

8. The method of claim 1 , wherein the polypeptide comprises the V H -PE38 subunit of SEQ ID NO: 1 and the V L subunit of SEQ ID NO: 2.

9. The method of claim 1 , wherein the composition comprises less than 20% of the deamidated species.

10. The method of claim 1 , wherein the composition comprises less than 10% of the deamidated species.

11. The method of claim 1 , wherein the composition comprises less than 5% of the deamidated species.

12. The method of claim 1 , wherein the composition comprises less than 3% of the deamidated species.

13. The method of claim 1 , wherein the composition comprises less than 2% of the deamidated species.

14. The method of claim 1 , wherein the composition comprises less than 1% of the deamidated species.

15. The method of claim 1 , wherein the composition further comprises a pharmaceutically acceptable carrier.

16. The method of claim 1 , wherein the composition comprises a formulation comprising 25 mM sodium phosphate, 4% sucrose, 8% glycine, and 0.02% polysorbate 80.

17. The method of claim 1 , wherein the composition is administered at a dose of 20-50 μg/kg per day.

18. The method of claim 1 , wherein the composition is administered by parenteral administration.

19. The method of claim 18 , wherein the composition is administered by intravenous administration.

Assignees

• Medimmune Ltd

Inventors

• Linke Thomas
• Wang William K
• Shah Ambarish
• Sathish Hasige
• Hunter Alan
• Thompson Christopher

Classifications

• B01D15/166 Primary

  Fluid composition conditioning, e.g. gradient · CPC title

• B01D15/363

  Anion-exchange · CPC title

• B01J41/14

  obtained by reactions only involving unsaturated carbon-to-carbon bonds · CPC title

• C07K17/02

  Peptides being immobilised on, or in, an organic carrier · CPC title

• B01J41/20

  Anion exchangers for chromatographic processes · CPC title