Method of weak partitioning chromatography
View Patent ↗This invention relates to methods of using weak partitioning chromatography for the purification of a product from a load fluid containing one or more impurities. Further, the invention relates to methods of weak partitioning chromatography defined by operating conditions which cause a medium to bind least 1 mg of product per mL of medium, or alternatively, defined by a partition coefficient of at least 0.1.
1. A method of recovering a purified product from a load fluid including one or more impurities, comprising the steps of:
passing the load fluid through a medium at operating conditions which cause the medium to bind at least 1 mg of product per mL of medium; and
recovering the purified product in the column effluent during the load cycle and any essentially isocratic wash.
2. A method of recovering a purified product from a load fluid including one or more impurities, comprising the steps of:
passing the load fluid through a medium at operating conditions defined by a partition coefficient of at least 0.1; and
recovering the purified product in the column effluent during the load cycle and any essentially isocratic wash.
3. A method of recovering a purified product from a load id including one or more impurities, comprising the steps of:
passing the load fluid through a medium at operating conditions which cause the medium to bind from at least 1 to about 70 mg of product per mL of medium and defined by a partition coefficient of 0.3 to 20; and
recovering the purified product in the column effluent during the load cycle and any essentially isocratic wash.
4. The method of claim 1 , wherein the operating conditions are identified in a screening step.
5. The method of claim 1 , wherein the operating conditions comprise pH levels and ionic strengths.
6. The method of claim 5 , wherein the operating conditions further comprise salt concentrations.
7. The method of claim 5 , wherein the operating conditions further comprise excipient concentrations.
8. The method of claim 7 , wherein the operating conditions further comprise phosphate concentrations, calcium concentrations, arginine concentrations, glycine concentrations, and HEPES concentrations.
9. The method of claim 1 , wherein the operating conditions comprise counterligand levels and pH levels.
10. The method of claim 9 , wherein the operating conditions further comprise imidazole concentrations.
11. The method of claim 4 , wherein the screening step uses batch binding studies.
12. The method of claim 4 , wherein the screening step uses column binding studies.
13. The method of claim 12 , wherein the column binding studies are gradient elution studies.
14. The method of claim 12 , wherein the column binding studies are isocratic elution studies.
15. The method of claim 1 , wherein the operating conditions cause the medium to bind at least 5 mg of product per mL of medium.
16. The method of claim 1 , wherein the operating conditions cause the medium to bind at least 10 mg of product per mL of medium.
17. The method of claim 1 , wherein the operating conditions cause the medium to bind at least 20 mg of product per mL of medium.
18. The method of claim 1 , wherein the operating conditions cause the medium to bind at least 30 mg of product per mL of medium.
19. The method of claim 1 , wherein the operating conditions cause the medium to bind at least 40 mg of product per mL of medium.
20. The method of claim 1 , wherein the operating conditions cause the medium to bind at least 50 mg of product per mL of medium.