Process of affinity chromatography
AssigneeKashiv Biosciences, LLC
InventorsOm Narayan, Tarun Kumar Gupta, Mayankkumar Thakkar
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Abstract
A process for purification of antibody or fusion protein by affinity chromatography wherein the elution is performed with high salt concentration which reduce turbidity in protein mixture during neutralization steps. The present invention provides an improved process of purifying antibodies through affinity chromatography using high salt-based elution.
Claims18
A process of purifying a protein mixture, wherein the process consisting of:
- a) equilibrating the affinity column with an equilibration buffer, wherein the equilibration buffer has a pH of about 6.5 to about 7.5;
- b) loading the protein mixture onto the affinity column in an amount of about 10 g/l to about 50 g/l;
- c) washing the affinity column with a first wash buffer at a pH of about 6.5 to about 7.5;
- d) optionally performing second and third wash, at a pH of about 6.5 to about 7.5;
- e) eluting the protein mixture with an elution buffer, wherein the elution buffer has a concentration of about 100 mM to about 200 mM and a pH of 3.0 to about 3.5;
- f) performing a viral inactivation and neutralization of eluted protein mixture to obtain a neutralized protein mixture.
A process of purifying a protein mixture, the protein mixture comprises omalizumab, and impurities, wherein the process consisting essentially of the following steps:
- a) equilibrating the affinity column with an equilibration buffer, wherein the equilibration buffer has a pH of about 6.5 to about 7.5;
- b) loading the protein mixture onto the affinity column in an amount of about 10 g/l to about 50 g/l;
- c) washing the affinity column with a first wash buffer at a pH of about 6.5 to about 7.5;
- d) optionally performing second and third wash at a pH of about 6.5 to about 7.5;
- e) eluting the protein mixture with an elution buffer, wherein the elution buffer has a concentration of about 100 mM to about 200 mM and a pH of 3.0 to about 3.5;
- f) performing a viral inactivation and neutralization of eluted protein mixture to obtain a neutralized protein mixture.
Dependencies are inferred from the claim text.