Method for chromatography reuse

Patent metadata

Field	Value
Publication number	US-10940401-B2
Application number	US-201815920237-A
Country	US
Kind code	B2
Filing date	Mar 13, 2018
Priority date	Sep 5, 2013
Publication date	Mar 9, 2021
Grant date	Mar 9, 2021

Abstract

The present invention provides methods for cleaning or regenerating a chromatography materiel for reuse. The methods of the invention can be used for cleaning or regenerating chromatography columns for reuse in the large-scale manufacture of multiple polypeptide products.

First claim

What is claimed is:

1. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing two or more material volumes of elution buffer through the material, wherein the elution buffer comprises about 0.15 M acetic acid and is about pH 2.9;
b) statically holding the material in elution buffer for a time ranging from about 10 minutes to about 30 minutes;
c) passing about two or more material volumes of elution buffer through the material; and
d) passing about two or more material volumes of regeneration buffer through the material, wherein the regeneration buffer comprises about 0.1 N NaOH and is about pH 13;
wherein the buffers are passed through the material at about 15 material volumes/hour.

2. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing about two material volumes of elution buffer through the material, wherein the elution buffer comprises about 0.15 M acetic acid and is about pH 2.9;
b) statically holding the material in elution buffer for about 30 minutes;
c) passing about two material volumes of elution buffer through the material; and
d) passing about four material volumes of regeneration buffer through the material, wherein the regeneration buffer comprises about 0.1 N NaOH and is about pH 13;

wherein the buffers are passed through the material at about 15 material volumes/hour.

3. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing about two material volumes of elution buffer through the material, wherein the elution buffer comprises about 0.15 M acetic acid and is about pH 2.9,
b) statically holding the material in elution buffer for about 30 minutes,
c) passing about two material volumes of elution buffer through the material, and
d) passing about two and one-half material volumes of regeneration buffer through the material, wherein the regeneration buffer comprises about 0.1 N NaOH and is about pH 13,
e) statically holding the material in regeneration buffer for about 30 minutes,
f) passing about two and one-half material volumes of regeneration buffer through the material;
wherein the buffers are passed through the material at about 15 material volumes/hour.

4. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing about two material volumes of equilibration buffer through the material, wherein the equilibration buffer comprises about 25 mM Tris and about 25 mM NaCl and is about pH 7.1;
b) statically holding the material in equilibration buffer for about 30 minutes;
c) passing about two material volumes of equilibration buffer through the material;
d) passing about two material volumes of elution buffer through the material, wherein the elution buffer comprises about 0.15 M Acetic acid and is about pH 2.8;
e) statically holding the material in elution buffer for about 30 minutes;
f) passing about two material volumes of elution buffer through the material;
g) passing about two material volumes of regeneration buffer through the material, wherein the regeneration buffer comprises 0.1 N NaOH, pH 13;
h) statically holding the material in regeneration buffer for about 30 minutes;
i) passing about two material volumes of regeneration buffer through the material.

5. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing about four material volumes of equilibration buffer through the material, wherein the equilibration buffer comprises about 25 mM Tris and about 25 mM NaCl and is pH 7.1;
b) performing six cycles of the steps comprising
i) passing about three material volumes of elution buffer through the material, wherein the elution buffer comprises about 0.15 M Acetic acid and is about pH 2.8;
ii) statically holding the material in elution buffer for about 10 minutes;
iii) passing about one material volume of elution buffer through the material;
iv) passing about three material volumes of regeneration buffer through the material, wherein the regeneration buffer comprises about 0.1 N NaOH and is about pH 13;
v) statically holding the material in regeneration buffer for about 10 minutes;

vi) passing about one material volume of regeneration buffer through the material.

6. A method to clean a chromatography material for reuse, the method comprising six cycles of the steps of
a) passing about three material volumes of elution buffer through the material, wherein the elution buffer comprises about 0.15 M Acetic acid and is about pH 2.8;
b) statically holding the material in elution buffer for about 15 minutes;
c) passing about one material volume of elution buffer through the material;
d) passing about three material volumes of regeneration buffer through the material, wherein the regeneration buffer comprises about 0.1 N NaOH and is about pH 13;
e) statically holding the material in regeneration buffer for about 15 minutes;
f) passing about one material volume of regeneration buffer through the material;
g) passing about three material volumes of storage buffer through the material, wherein the storage buffer comprises about 100 mM sodium acetate, about 2% benzyl alcohol, and is about pH 5.0;
e) statically holding the material in storage buffer for about 15 minutes;
f) passing about one material volume of storage buffer through the material;
wherein the buffers are passed through the material at about 15 material volumes/hour.

7. The method of claim 5 , wherein the chromatography material is in a chromatography column.

8. The method of claim 5 , wherein the chromatography material is an affinity material.

9. The method of claim 8 , wherein the affinity material is a protein A affinity material.

10. The method of claim 5 , wherein the chromatography material is used for large-scale production of a polypeptide.

11. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing about three material volumes of equilibration buffer through the material, wherein the equilibration buffer comprises about 40 mM sodium acetate and is about pH 5.5;
b) passing about two material volumes of about 0.5 N NaOH through the material
c) statically holding the material in about 0.5 N NaOH for about 10 minutes;
d) passing about one material volume of about 0.5 N NaOH through the material; and
e) statically holding the material in about 0.5 N NaOH for about 10 minutes;
f) passing about one material volume of about 0.5 N NaOH through the material.

12. The method of claim 11 , wherein the chromatography material is an ion exchange material.

13. The method of claim 12 herein the ion exchange material is a cation exchange material.

14. The method of claim 11 , wherein the chromatography material is used for large-scale production of an antibody.

15. A method to clean a chromatography material for reuse, the method comprising the steps of
a) passing about three material volumes of equilibration buffer through the material, wherein the equilibration buffer comprises about 50 mM Tris, 85 mM sodium acetate and is about pH 8.8 and about 8.6 mS/cm;
b) passing about two material volumes of about 0.5 N NaOH through the material
c) statically holding the material in about 0.5 N NaOH for about 10 minutes;
d) passing about one material volume of about 0.5 N NaOH through the material; and
e) statically holding the material in about 0.5 N NaOH for about 10 minutes;
f) passing about one material volume of about 0.5 N NaOH through the material.

16. The method of claim 15 , wherein th

Assignees

• Genentech Inc

Inventors

• Mahajan Ekta
• So Joanna
• Werber Jay

Classifications

• C07K1/22

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

• B01D15/3809

  of the antigen-antibody type, e.g. protein A, G or L chromatography · CPC title

• C07K1/18

  Ion-exchange chromatography · CPC title

• B01D15/203 Primary

  Equilibration or regeneration · CPC title

• B01J49/60

  Cleaning or rinsing ion-exchange beds · CPC title