Method for purifying antibodies using PBS

Patent metadata

Field	Value
Publication number	US-10329323-B2
Application number	US-201514809211-A
Country	US
Kind code	B2
Filing date	Jul 25, 2015
Priority date	Jul 25, 2014
Publication date	Jun 25, 2019
Grant date	Jun 25, 2019

Abstract

Disclosed here includes a method for purifying a biologic composition, comprising diafiltering the biologic composition into a composition comprising phosphate buffered saline (PBS) to obtain a purified composition. The method disclosed here can be particularly useful for removing one or more impurities from the biologic composition, such as bis(2-hydroxyethyl)amino-tris(hydroxymethyl)methane (Bis-tris).

First claim

What is claimed is:

1. A method for purifying a monoclonal antibody, comprising:
(a) passing a first composition comprising the monoclonal antibody through an affinity chromatography column to obtain a second composition comprising the monoclonal antibody;
(b) lowering the pH of the second composition to obtain a third composition;
(c) washing the third composition with a solvent-detergent to obtained a fourth composition;
(d) washing the fourth composition to remove the solvent-detergent to obtain a fifth composition;
(e) passing the fifth composition through a cation exchange chromatography column to obtain a sixth composition comprising the monoclonal antibody;
(f) subjecting the sixth composition to nano-filtration to obtain a seventh composition;
(g) passing the seventh composition through a mixed mode chromatography column to obtain an eighth composition comprising the monoclonal antibody and Bis-tris; and
(h) diafiltering the eighth composition into a composition comprising PBS to obtain a ninth composition comprising the monoclonal antibody but substantially free from Bis-tris,
wherein the monoclonal antibody is ch14.18.

2. The method of claim 1 , wherein the concentration of the Bis-tris in step (g) in the biologic composition is 10 to 50 mM Bis-tris at a pH of 6.3 to 6.7.

3. The method of claim 1 , wherein the diafiltering of step (h) removes at least 50% of the Bis-tris from the biologic composition.

4. The method of claim 1 , wherein the diafiltering of step (h) removes at least 70% of the Bis-tris from the biologic composition.

5. The method of claim 1 , wherein the concentration of the PBS in step (h) is 10 to 50 mM Sodium Phosphate and 100 to 200 mM NaCl.

6. The method of claim 1 , wherein the monoclonal antibody is concentrated to a concentration before being diafiltered into the composition comprising PBS in step (h).

7. The method of claim 1 , wherein the mixed mode chromatography column of step (g) has a resin with a combination of anion exchange and hydrophobic interaction.

8. The method of claim 7 , wherein the monoclonal antibody is eluted in step (g) from the mixed mode chromatography column with a combination of anion exchange and hydrophobic interaction using a composition comprising Bis-tris.

9. The method of claim 1 , wherein at least three volume units of the eighth composition is diafiltered into one volume unit of the composition comprising PBS in step (h).

10. The method of claim 1 , wherein at least five volume units of the eighth composition is diafiltered into one volume unit of the composition comprising PBS in step (h).

Assignees

• The Us Secretary Department Of Health & Human Services
• United Therapeutics Corp

Inventors

• Shaban Samir H
• Koleck Mary P
• Meh David A
• Farquharson Gerald M
• Atolagbe Timothy O
• Mitra George

Classifications

• B01D15/1871

  placed in series · CPC title

• B01D15/24

  relating to the treatment of the fractions to be distributed · CPC title

• C07K1/36 Primary

  by a combination of two or more processes of different types · CPC title

• B01D15/125

  Pre-filtration · CPC title

• C07K1/34

  by filtration, ultrafiltration or reverse osmosis · CPC title