Purification method which prevents denaturation of an antibody
The present invention provides a method of purifying an antibody by protein A affinity chromatography. More specifically, the present invention provides a technique relating to an elution buffer solution which provides a good antibody recovery rate without denaturation.
1. A method of purifying an antibody comprising
a) placing a starting material containing said antibody on a protein A affinity chromotography column,
b) desorbing said antibody from said column using a buffer solution adjusted to pH 4.0 to 5.0 comprising arginine and/or a derivative thereof, and
c) recovering the antibody, wherein the concentration of said arginine and/or derivative thereof provides an antibody recovery rate equivalent to that of pH 3.5 sodium citrate buffer solution.
2. The method of claim 1 , wherein said antibody recovery does not induce association or aggregation of the antibody.
3. The method of claim 1 , wherein said starting material is selected from the group consisting of a monoclonal antibody solution, a mouse myeloma cell solution, a humanized antibody-producing cell solution and a human-type antibody-producing cell solution.
4. The method of claim 1 , wherein said starting material is humanized antibody-producing cells, human-type antibody-producing cells, a humanized antibody-producing cell solution and a human-type antibody-producing cell solution.
5. The method of claim 1 , wherein said starting material is CHO cells.
6. The method of claim 1 , wherein said buffer solution is adjusted to pH 4.3 to 4.7.
7. The method of claim 1 , wherein the concentration of said arginine and or arginine derivative is from 0.1 to 4.0 M.
8. The method of claim 7 , wherein the concentration of said arginine and or arginine derivative is from 1 to 2 M.
9. The method of claim 1 , wherein said antibody is selected from the group consisting of a natural human antibody, a humanized antibody, a human-type antibody, an antibody prepared by genetic recombination and a monoclonal antibody.
10. The method of claim 1 , wherein said antibody is selected from the group consisting of a natural human antibody, a humanized antibody and a human-type antibody.
11. A method of purifying an antibody comprising
a) placing a starting material containing said antibody on a protein A affinity chromotography column,
b) desorbing said antibody from said column using a buffer solution adjusted to pH 4.0 to 5.0 comprising arginine and/or an arginine derivative, and
c) recovering the antibody,
wherein said arginine derivative is selected from the group consisting of acetyl arginine, agmatine, L-arginic acid, N-α-butyroyl-L-arginine, and N-α-pyvaloyl arginine.
12. The method of claim 11 , wherein said arginine derivative is N-α-butyroyl-L-arginine.
13. A method of producing a purified antibody comprising:
a) placing a starting material containing an antibody on a protein A affinity chromotography column,
b) desorbing said antibody from said column using a buffer solution adjusted to pH 4.0 to 5.0 comprising arginine and/or an arginine derivative, and
c) recovering the antibody,
wherein said arginine derivative is selected from the group consisting of acetyl arginine, agmatine, L-arginic acid, N-α-butyroyl-L-arginine, and N-α-pyvaloyl arginine, wherein the concentration of said arginine and/or derivative thereof provides an antibody recovery rate equivalent to that of pH 3.5 sodium citrate buffer solution.
14. The method of claim 11 , wherein said arginine derivative is N-α-butyroyl-L-arginine.