Surface neutralization of apatite

Patent metadata

Field	Value
Publication number	US-10676502-B2
Application number	US-201815879239-A
Country	US
Kind code	B2
Filing date	Jan 24, 2018
Priority date	Jan 15, 2010
Publication date	Jun 9, 2020
Grant date	Jun 9, 2020

Abstract

The present invention discloses methods of neutralizing apatite surfaces, for example during chromatography and before protein elution.

First claim

What is claimed is:

1. A mixture comprising:
i) an aqueous solution comprising:
a) 5 mM to 50 mM of a sulphonated amine compound;
b) 100 mM or less of an alkali metal ion, or
an alkali earth metal ion; and
c) a pH of 6.5 to 9;
ii) a target molecule; and
iii) an apatite solid surface,
wherein said apatite solid surface is in contact with the aqueous solution, and at least 90% of said target molecule is adsorbed onto the apatite solid surface.

2. The mixture of claim 1 , wherein the target molecule is a protein.

3. The mixture of claim 2 , wherein the target molecule comprises an antibody, a monoclonal antibody, a human antibody, a humanized antibody, or a fragment thereof.

4. The mixture of claim 3 , wherein the mixture comprises serum albumin, and wherein said serum albumin is dissolved in the aqueous solution.

5. The mixture of claim 1 , wherein the apatite is selected from the group consisting of hydroxyapatite and fluorapatite.

6. The mixture of claim 1 , wherein the apatite is ceramic hydroxyapatite or ceramic fluorapatite.

7. The mixture of claim 1 , wherein the apatite is a non-ceramic apatite.

8. The mixture of claim 1 , wherein the aqueous solution comprises a pH of from 7 to 9.

9. The mixture of claim 1 , wherein the aqueous solution has from 25 mM to 100 mM of the alkali metal ion.

10. The mixture of claim 1 , wherein the aqueous solution has 60 mM or less of alkali earth metal ion.

11. The mixture of claim 10 , wherein the alkali earth metal ion is a calcium ion.

12. The mixture of claim 11 , wherein the calcium ion of the aqueous solution has a concentration consisting of 0.25 mM or less.

13. The mixture of claim 1 , wherein the aqueous solution comprises 25 mM Tris-HCL, 25 mM NaCl, pH 8.4.

14. The mixture of claim 1 , wherein the aqueous solution comprises phosphate ions.

15. The mixture of claim 14 , wherein the phosphate ions are at a concentration of 5 mM.

16. The mixture of claim 1 , wherein the aqueous solution has a volume consisting of at least 4 times a volume of the apatite solid surface.

17. The mixture of claim 1 , wherein the sulphonated amine compound is selected from PIPES, MES, MOPS, ACES, MOPSO, and HEPES.

18. The mixture of claim 1 , wherein the concentration of the sulphonated amine compound is between 5-100 mM.

19. The mixture of claim 18 , wherein the sulphonated amine compound is selected from PIPES, MES, MOPS, ACES, MOPSO, and HEPES.

Assignees

• Bio Rad Laboratories

Inventors

• Cummings Larry J
• Snyder Mark A

Classifications

• B01J20/281

  Sorbents specially adapted for preparative, analytical or investigative chromatography · CPC title

• B01J20/048

  containing phosphorus, e.g. phosphates, apatites, hydroxyapatites · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title

• B01D15/3847

  Multimodal interactions · CPC title

• B01D15/424 Primary

  Elution mode · CPC title