Methods for inactivating viruses during a protein purification process
View Patent ↗The present application relates to novel and improved methods of achieving virus inactivation during a protein purification process.
1. A method of inactivating one or more viruses in a Protein A eluate as the Protein A eluate flows from a Protein A affinity chromatography step to a flow-through purification step, the method comprising mixing the Protein A eluate with one or more virus inactivating agents using one or more in-line static mixers, wherein the flow of the Protein A eluate through the static mixer is in the laminar flow range comprising a Reynolds number below 100, wherein complete virus inactivation is achieved in less than 10 minutes or less than 5 minutes or less than 2 minutes or less than 1 minute.
2. A method for inactivating one or more viruses, the method comprising:
a. subjecting a sample comprising a target protein to a Protein A affinity chromatography process, thereby to obtain an eluate; and
b. continuously transferring the eluate to an in-line static mixer to mix one or more virus inactivating agents with the eluate for a duration of time which is equal to or less than 10 minutes, wherein the flow of the eluate through the static mixer is in the laminar flow range comprising a Reynolds number below 100, thereby to inactivate one or more viruses.
3. The method of claim 2 , wherein the Protein A affinity chromatography process is performed in batch mode.
4. The method of claim 2 , wherein the Protein A affinity chromatography process is performed in a continuous mode.
5. The method of claim 4 , wherein the process comprises continuous multi-column chromatography.
6. The method of claim 2 , wherein the one or more virus inactivating agents is an acid.
7. The method of claim 2 , wherein the target protein is an antibody.
8. The method of claim 2 , further comprising the step of continuously transferring output from step (b) into a flow-through purification process step.
9. The method of claim 8 , wherein the flow-through purification step comprises two or more matrices selected from activated carbon, anion exchange media, cation exchange media and virus filter.
10. The method of claim 1 , wherein the flow-through purification step comprises two or more matrices selected from activated carbon, anion exchange media, cation exchange media and virus filter.