Method for purifying active polypeptides or immunoconjugates

Patent metadata

Field	Value
Publication number	US-10072083-B2
Application number	US-201715423928-A
Country	US
Kind code	B2
Filing date	Feb 3, 2017
Priority date	Jul 30, 2010
Publication date	Sep 11, 2018
Grant date	Sep 11, 2018

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 composition comprising a purified polypeptide comprising an anti-CD22 antibody or antigen binding fragment thereof and a Pseudomonas exotoxin (PE) or variant thereof 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 composition 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.

3. The composition of claim 1 , wherein the PE or variant thereof has the amino acid sequence of SEQ ID NO: 22.

4. The composition of claim 1 , wherein the antibody or antigen binding fragment thereof comprises a VH and a VL sequence.

5. The composition of claim 4 , wherein the VH sequence is selected from the group consisting of SEQ ID NOs: 6-11.

6. The composition of claim 4 , wherein the VL sequence is selected from the group consisting of SEQ ID NOs: 2 and 12-15.

7. The composition 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.

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

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

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

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

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

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

14. A pharmaceutical composition comprising the composition of claim 1 and a pharmaceutically acceptable carrier.

15. A unit dosage form of a purified polypeptide in the range of 0.1 mg to 6 mg, wherein the polypeptide comprises an anti-CD22 antibody or antigen binding fragment thereof and a PE or variant thereof having an amino acid sequence selected from the group consisting of SEQ ID NOs: 16-22, wherein the unit dosage form comprises less than 25% deamidated species of the polypeptide.

16. The unit dosage form of claim 15 , wherein the polypeptide comprises the V H -PE38 subunit of SEQ ID NO: 1 and the V L subunit of SEQ ID NO: 2.

17. A formulation comprising the composition of claim 1 and at least one excipient selected from the group consisting of sodium chloride, potassium dihydrogen phosphate, disodium hydrogen phosphate, sodium hydroxide, and water.

18. The unit dosage form of claim 15 , wherein the PE or variant thereof has the amino acid sequence of SEQ ID NO: 22.

19. The composition of claim 1 , wherein the anti-CD22 antibody or antigen binding fragment thereof and PE or variant thereof is at a concentration of 1.0 mg/mL at pH 7.4 and further comprises 25 mM sodium phosphate, 4% sucrose, 8% glycine, and 0.02% polysorbate 80.

20. The composition of claim 1 , wherein the formulation is lyophilized.

21. An isolated polypeptide comprising the V H -PE38 subunit of SEQ ID NO: 1 and the V L subunit of SEQ ID NO: 2.

Assignees

• Medimmune Llc

Inventors

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

Classifications

• A61P39/02

  Antidotes · CPC title

• A61K47/02

  Inorganic compounds · CPC title

• C07K2319/55

  containing a fusion with a toxin, e.g. diphteria toxin · CPC title

• B01J41/20

  Anion exchangers for chromatographic processes · CPC title

• C07K2317/94

  Stability, e.g. half-life, pH, temperature or enzyme-resistance · CPC title