Method for protein purification
The present invention provides a method for manufacturing antibodies or a fragment thereof with reduced levels of antibody reduction related impurities.
1. A method for manufacturing an antibody or fragment thereof comprising:
expressing said antibody or fragment thereof in a host cell;
recovering a mixture containing the antibody or fragment thereof, host cell debris, and impurities; and
purifying said antibody or fragment thereof from the mixture wherein said purification comprises:
activated carbon filtration, followed by
protein A chromatography.
2. The method according to claim 1 , wherein said purification comprises at least one further chromatography step.
3. The method according to claim 2 , wherein said further chromatography steps comprise cation exchange chromatography and/or anion exchange chromatography.
4. The method according to claim 1 , wherein the recovered antibody or fragment thereof comprises native disulfide bonds.
5. The method according to claim 1 , wherein the recovered antibody or fragment thereof comprises native inter-chain disulfide bonds.
6. The method according to claim 1 , wherein said host cell is a mammalian host cell and the mixture containing the antibody is the cell culture supernatant.
7. The method according to claim 6 , wherein said purification process additionally comprises depth filtration of the cell culture supernatant prior to activated carbon filtration.
8. The method according to claim 2 , wherein depth filtration and activated carbon filtration are operated in-line.
9. The method according to claim 1 , wherein the antibody or fragment recovered from the protein A chromatography comprises less than 10%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1% antibody reduction related impurities.
10. The method according to claim 1 , wherein the antibody or fragment recovered from the protein A chromatography comprises less antibody reduction related impurities than the antibody or fragment recovered from the same process without an activated carbon filtration step.
11. A method for the prevention of disulfide bond reduction during purification of an antibody or fragment thereof expressed in a host cell comprising:
activated carbon filtration step, followed by
protein A chromatography.
12. A method for purifying an antibody or fragment thereof from a liquid mixture containing contaminants comprising:
filtering said liquid mixture with an activated carbon filter, followed by
protein A chromatography.
13. The method according to claim 12 , wherein said recovered antibody or fragment thereof comprises native disulfide bonds.
14. The method according to claim 13 , wherein the recovered protein comprises native inter-chain disulfide bonds.
15. A method for decreasing the amount of reducing agents during purification of an antibody or antibody fragment thereof comprising:
activated carbon filtration, followed by
protein A chromatography.