Hydrophobic interaction protein chromatography under no-salt conditions

• US 9,994,609 B2
• Filed: 03/15/2014
• Issued: 06/12/2018
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Biogen MA Inc. (Biogen, Inc.)
• Original Assignee: Biogen MA Inc. (Biogen, Inc.)
• Inventors: Ghose, Sanchayita, Tao, Yinying
• Primary Examiner(s): Kolker, Daniel E
• Assistant Examiner(s): Rogers, James L
• Application Number:
  US14/775,774
• Publication Number:
  US 20160024145A1
• Time in Patent Office: 1,550 Days
• Field of Search: None
• CPC Class Codes: C07K 1/20 Partition-, reverse-phase or hydrophobic interaction chromatography
• CPC Class Codes: C07K 16/065 Purification, fragmentation

Abstract

Various aspects and embodiments of the present disclosure relate to the purification antibodies by hydrophobic interaction chromatography under no-salt conditions.

Claims

1. A method of purifying an antibody, the method comprising:

subjecting an antibody in solution to hydrophobic interaction chromatography in flow through mode using a matrix comprising a hydroxylated methacrylic polymer containing hexyl groups, wherein the solution does not contain an ammonium sulfate salt and has a pH of about 5.0 to about 7.0.

2. The method of claim 1, wherein matrix has a binding capacity for lysozyme of between 30 mg/ml and 50 mg/ml.

3. The method of claim 1, wherein the pH of the solution is about 5.0 to about 6.5, about 5.0 to about 6.0, about 5.5 to about 7.0, or about 6.0 to about 7.0.

4. The method of claim 1, wherein the antibody is a monoclonal antibody or a polyclonal antibody.

5. The method of claim 4, wherein the antibody is a human antibody, a mouse antibody or a chimeric antibody.

6. A method of purifying an antibody, the method comprising:

providing an antibody in a solution that does not contain an ammonium sulfate salt and has a pH of about 5.0 to about 7.0;

loading the solution onto a matrix comprising a hydroxylated methacrylic polymer containing hexyl groups; and

collecting a flow through fraction that contains the antibody.

7. The method of claim 6, wherein matrix has a binding capacity for lysozyme of between 30 mg/ml and 50 mg/ml.

8. A method of purifying an antibody, the method comprising:

subjecting an antibody in solution to hydrophobic interaction chromatography in flow through mode using a hydroxylated methacrylic polymer matrix containing hexyl groups, wherein the solution does not contain a kosmotropic salt and has a pH of about 5.0 to about 7.0.

9. The method of claim 8, wherein the pH of the solution is about 5.0 to about 6.5, about 5.0 to about 6.0, about 5.5 to about 7.0, or about 6.0 to about 7.0.

10. The method of claim 8, wherein the antibody is a monoclonal antibody or a polyclonal antibody.

11. The method of claim 10, wherein the antibody is a human antibody, a mouse antibody or a chimeric antibody.

12. The method of claim 8, wherein matrix has a binding capacity for lysozyme of between 30 mg/ml and 50 mg/ml.

13. A method of purifying an antibody, the method comprising:

providing an antibody in a solution that does not contain a kosmotropic salt and has a pH of about 5.0 to about 7.0;

loading the solution onto a hydroxylated methacrylic polymer matrix containing hexyl groups; and

collecting a flow through fraction that contains the antibody.

14. The method of claim 13, wherein matrix has a binding capacity for lysozyme of between 30 mg/ml and 50 mg/ml.

15. The method of claim 13, wherein the pH of the solution is about 5.0 to about 6.5, about 5.0 to about 6.0, about 5.5 to about 7.0, or about 6.0 to about 7.0.

16. The method of claim 13, wherein the antibody is a monoclonal antibody or a polyclonal antibody.

17. The method of claim 16, wherein the antibody is a human antibody, a mouse antibody or a chimeric antibody.