Methods of inactivating viral contaminants
The present invention relates to a method for preparing an antibody-containing solution free of viral contaminants starting from cultured cells are described. The method include a step of subjecting the antibody containing solution to a mix of solvent and detergent or to high pH.
1. A method for the removal of impurities from a clarified harvest antibody material, the method comprising: (a) culturing host cells which express an antibody, (b) harvesting the antibody material in the cell culture and (c) subjecting it to a clarification step, wherein said method further comprises (d) treating the resulting clarified harvest antibody material with high pH to reach a high target pH and (e) subjecting the high pH treated clarified harvest antibody material to a filtration step to remove impurities comprising host cell proteins, nucleic acids and/or cell debris wherein said high target pH is about 10.2 to about 11.0.
2. The method of claim 1 , wherein said high target pH is about 10.8.
3. The method of claim 1 , wherein said high target pH is about 11.
4. The method of claim 1 , wherein said high target pH is reached using a buffer selected from the group consisting of: Tris, Histidine L-Arginine, phosphate and NaOH.
5. The method of claim 1 , wherein said high target pH is about 10.2.
6. The method of claim 1 , wherein said high target pH is about 10.4.
7. The method of claim 1 , wherein the resulting clarified harvest antibody material is treated with pH of about 10.5 to about 11.0.
8. The method of claim 1 , wherein the resulting clarified harvest antibody material is treated with pH of about 10.8.
9. A method for the removal of impurities from a clarified harvest antibody material, the method comprising: (a) culturing host cells which express an antibody, (b) harvesting the antibody material in the cell culture and (c) subjecting it to a clarification step, wherein said method further comprises (d) subjecting said clarified harvest antibody material to protein A chromatography, (e) treating a resulting protein A eluate with high pH to reach a high target pH and (f) subjecting the high pH treated protein A eluate to a filtration step to remove impurities comprising host cell proteins, nucleic acids and/or cell debris, wherein said high target pH is about 10.2 to about 11.0.
10. The method of claim 9 , wherein step (e) comprises treating the resulting protein A eluate with said high pH to reach said high target pH and then neutralizing the high target pH protein A eluate.
11. The method of claim 9 , wherein in step (e) treating the resulting protein A eluate with said high pH comprises incubating the resulting protein A eluate with NaOH 0.5M to reach a high target pH of about 11 and further comprises holding the protein A eluate at the high target pH for at least 60 min at room temperature.
12. The method of claim 9 , wherein in step (e) treating the resulting protein A eluate with said high pH comprises incubating the resulting protein A eluate with NaOH 0.5M to reach a high target pH of about 11 and further comprises holding the protein A eluate at the high target pH for at least 60 min at room temperature, and then neutralizing the high target pH protein A eluate.
13. The method of claim 9 , wherein in step (e) treating the resulting protein A eluate with said high pH comprises incubating the resulting protein A eluate with NaOH 0.5M to reach a high target pH 11.2 and further comprises holding the protein A eluate at the high target pH for at least 60 min at room temperature.
14. The method of claim 9 , wherein in step (e) treating the resulting protein A eluate with said high pH comprises incubating the resulting protein A eluate with NaOH 0.5M to reach a high target pH 11.2 and further comprises holding the protein A eluate at the high target pH for at least 60 min at room temperature, and then neutralizing the high target pH protein A eluate.