Method for purifying active polypeptides or immunoconjugates

Patent metadata

Field	Value
Publication number	US-2017145100-A1
Application number	US-201715423928-A
Country	US
Kind code	A1
Filing date	Feb 3, 2017
Priority date	Jul 30, 2010
Publication date	May 25, 2017
Grant date	—

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, wherein the composition comprises less than between about 25% and about 1% 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 Pseudomonas exotoxin or variant thereof has an amino acid sequence selected from the group consisting of SEQ ID NOs: 16-22.

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

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

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

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

8 . 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.

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

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

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

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

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

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

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

16 . 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 Pseudomonas exotoxin (PE) or variant thereof, wherein the unit dosage form comprises less than between about 25% and about 1% deamidated species of the polypeptide.

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

18 . 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, and sodium hydroxide and water.

Assignees

• Medimmune Llc

Inventors

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

Classifications

• A61P39/02

  Antidotes · CPC title

• C07K2319/55

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

• A61K47/02

  Inorganic compounds · CPC title

• A61K47/6849

  the antibody targeting a receptor, a cell surface antigen or a cell surface determinant · CPC title

• B01D15/363

  Anion-exchange · CPC title