Process for reducing ligand leakage from affinity chromatography matrices
AssigneeBoehringer Ingelheim Pharma GmbH & Co. KG
InventorsLothar Halmer, Margit Holzer
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Abstract
The present invention relates to a process for (pre)treating affinity chromatography matrices, preferably protein A matrices, with at least one surfactant for reducing ligand leakage. By treating the affinity chromatography matrix according to the invention it is possible to achieve an affinity chromatography matrix of constant quality in terms of ligand leakage, which is an important prerequisite for its suitability or use in biopharmaceutical processes. The present invention further relates to methods for determining the ligand leakage.
Claims52
A process for treating an affinity chromatography matrix in order to reduce the ligand leakage comprising treating the affinity chromatography matrix with at least one surfactant, wherein the duration of treatment with the surfactant is at least 4 hours.
A process for treating an affinity chromatography matrix comprising the steps of:
- a. pre-treating the affinity chromatography matrix comprising:
- i. treating the affinity chromatography matrix with a chaotropic substance;
- ii. treating the affinity chromatography matrix with an acidic elution buffer having a pH of about 2.0 to about 4.0;
- and
- iii. treating the affinity chromatography matrix with a neutral equilibration buffer having a pH of about 6.5 to about 8.5;
- b. treating the affinity chromatography matrix with 5 to 15 bed volumes of at least one surfactant for at least 4 hours at a process temperature ranging from about 25° C. to 55° C.;
- and, optionally,
- c. post-treating the affinity chromatography matrix comprising:
- i. treating the affinity chromatography matrix with a chaotropic substance;
- ii. treating the affinity chromatography matrix with an acidic elution buffer having a pH of about 2.0 to about 4.0;
- and
- iii. treating the affinity chromatography matrix with a neutral equilibration buffer having a pH of about 6.5 to about 8.5.
A process for treating an affinity chromatography matrix comprising the steps of:
- a. pre-treating the affinity chromatography matrix comprising:
- i. treating the affinity chromatography matrix with a chaotropic substance;
- ii. treating the affinity chromatography matrix with an acidic elution buffer having a pH of about 2.0 to about 4.0;
- and
- iii. treating the affinity chromatography matrix with a neutral equilibration buffer having a pH of about 6.5 to about 8.5;
- b. treating the affinity chromatography matrix with 5 to 15 bed volumes of at least one surfactant for at least 16 hours at a process temperature less than 25° C.;
- and, optionally,
- c. post-treating the affinity chromatography matrix comprising:
- i. treating the affinity chromatography matrix with a chaotropic substance;
- ii. treating the affinity chromatography matrix with an acidic buffer having a pH of about 2.0 to about 4.0;
- and
- iii. treating the affinity chromatography matrix with a neutral equilibration buffer having a pH of about 6.5 to about 8.5.
A process for treating an affinity chromatography matrix comprising treating the affinity chromatography matrix with a solution comprising:
- (a) a chaotropic substance;
- (b) an acidic elution buffer having a pH of about 2.0 to about 4.0;
- (c) a neutral equilibration buffer having a pH of about 6.5 to about 8.5;
- and (d) 5 to 15 bed volumes of at least one surfactant, wherein the affinity chromatography matrix is treated for at least 4 hours at a process temperature ranging from about 25° C. to 55° C.
A process for treating an affinity chromatography matrix comprising treating the affinity chromatography matrix with a solution comprising:
- (a) a chaotropic substance;
- (b) an acidic buffer having a pH of about 2.0 to about 4.0;
- (c) a neutral equilibration buffer having a pH of about 6.5 to about 8.5;
- and (d) 5 to 15 bed volumes of at least one surfactant, wherein the affinity chromatography matrix is treated with 5 to 15 bed volumes of at least one surfactant for at least 16 hours at a process temperature less than 25° C.
Dependencies are inferred from the claim text.