Reducing pH excursions in ion exchange chromatography using displacing counter ions

Patent metadata

Field	Value
Publication number	US-9442050-B2
Application number	US-201313763064-A
Country	US
Kind code	B2
Filing date	Feb 8, 2013
Priority date	Feb 14, 2012
Publication date	Sep 13, 2016
Grant date	Sep 13, 2016

Abstract

Provided are methods and compositions for reducing or eliminating pH excursions in cation and anion exchange chromatography.

First claim

What is claimed is:

1. A method of suppressing formation of pH excursions on a charged solid support having associated hydrogen or hydroxide ions, the method comprising
providing a solid support in association with an equilibration buffer, a wash buffer, or a loading buffer and subsequently;
contacting the solid support with a solution comprising displacing counter ions with a charge opposite the net charge of the solid support, wherein the displacing counter ion is a positively-charged molecule thereby replacing hydrogen ions associated with a negatively charged solid support, or wherein the displacing counter ion is a negatively-charge molecule thereby replacing hydroxide ions associated with a positively charged solid support; and subsequently
eluting a target molecule from the solid support, wherein the eluting comprises increasing conductivity of an elution solution in contact with the solid support.

2. The method of claim 1 , wherein the solid support is a cation exchange support and the equilibration buffer has a pH from 4 to 7.

3. The method of claim 1 , wherein the solid support is an anion exchange support and the equilibration buffer has a pH from 6.5 to 9.

4. The method of claim 1 , wherein the solid support comprises negative charges and hydrogen ions are associated with the negative charges.

5. The method of claim 4 , wherein the solid support is a cation-exchange solid support.

6. The method of claim 4 , wherein the solid support is a mixed mode solid support having cation-exchange functionality.

7. The method of claim 6 , wherein the mixed mode solid support having cation-exchange functionality further comprises hydrophobic functionality.

8. The method of claim 5 , wherein the displacing counter ion is a positively-charged molecule selected from an amine, amine derivative, Tris, and histidine.

9. The method of claim 5 , wherein the displacing counter ion is at a concentration of about 100 to about 500 mM.

10. The method of claim 1 , wherein the solid support comprises positive charges and hydroxide ions are associated with the positive charges.

11. The method of claim 10 , wherein the solid support is an anion-exchange solid support.

12. The method of claim 10 , wherein the solid support is a mixed mode solid support having anion-exchange functionality.

13. The method of claim 12 , wherein the mixed mode solid support having anion-exchange functionality further comprises hydrophobic functionality.

14. The method of claim 10 , wherein the displacing agent is a negatively-charged molecule selected from glutamate, aspartate, phosphate, and citrate.

15. The method of claim 14 , wherein the displacing counter ion is at a concentration of about 100 to about 500 mM.

16. The method of claim 1 , wherein the eluting further comprises changing the pH of the elution solution.

17. The method of claim 1 , wherein the eluting does not substantially change the pH of the elution solution.

18. The method of claim 1 , wherein the solid support comprises negative charges associated with hydrogen ions and the eluting further comprises increasing the pH of the elution solution.

19. The method of claim 1 , wherein the solid support comprises positive charges associated with hydroxide ions and the eluting further comprises decreasing the pH of the elution solution.

20. The method of claim 1 , wherein the solution comprising displacing counter ions further comprises a buffer.

21. The method of claim 20 , wherein the buffer is a zwitterionic buffer.

22. The method of claim 21 , wherein the solid support comprises negative charges and the buffer is a negatively-charged buffer.

23. The method of claim 21 , wherein the solid support comprises positive charges and the buffer is a positively-charged buffer.

24. The method of claim 1 , wherein the target molecule is a biomolecule.

25. The method of claim 24 , wherein the biomolecule is a protein.

26. The method of claim 25 , wherein the protein is an antibody.

27. The method of claim 1 , wherein the positively-charged molecule is a buffering ion.

Assignees

• Gagnon Peter S
• Bio Rad Laboratories Inc

Inventors

• Gagnon Peter S

Classifications

• B01D15/327

  with hydrophobic interaction · CPC title

• B01D15/362

  Cation-exchange · CPC title

• G01N1/34 Primary

  Purifying; Cleaning {(processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003)} · CPC title

• Y10T436/25375

  Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.] · CPC title

• B01D15/422

  Displacement mode · CPC title