Chromatography equipment characterization

• US 8,357,301 B2
• Filed: 06/22/2010
• Issued: 01/22/2013
• Est. Priority Date: 06/24/2009
• Status: Active Grant

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Belousov, Anton, Dams, Thomas, Gerwat, Benjamin
• Primary Examiner(s): Therkorn, Ernest G.
• Application Number:
  US13/380,382
• Publication Number:
  US 20120202978A1
• Time in Patent Office: 945 Days
• Field of Search: 210/635, 210/656, 210/657, 210/659, 210/143, 210/198.2, 702/30, 702/81, 702/84, 702/104, 530/413
• US Class Current: 210/656
• CPC Class Codes: C07K 1/16 by chromatography
• CPC Class Codes: G01N 30/50 Conditioning of the sorbent material or stationary liquid
• CPC Class Codes: G01N 30/86 Signal analysis
• CPC Class Codes: G01N 30/88 Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86

Abstract

Herein is reported a method for determining whether a re-useable chromatography column packing, which is used at least for the second time in a purification step of a purification of a polypeptide, has reduced separation efficacy in said purification step of said purification of said polypeptide, comprising the following steps: a) identifying and determining the experimental data of an inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing, b) determining the parameters of a function of formula I by fitting the experimental data of the inert change of the physicochemical parameter of the at least second use, c) determining the difference between the experimental data of the inert change of the physicochemical parameter of the at least second use and the function of formula I with the parameters determined in step b), d) calculating the difference between the maximum value and the minimum value of the difference determined in step c) and normalizing said difference, e) determining reduced separation efficacy of said re-useable chromatography column packing when the absolute value of the difference calculated in step d) is more than 0.1.

Claims

1. A method for determining whether a re-useable chromatography column packing, which is used at least for the second time in a purification step of a purification of a polypeptide, has reduced separation efficacy in said purification step of said purification of said polypeptide, wherein said method comprises the following steps:

a) identifying and determining the experimental data of an inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing of the at least second use,

b) determining the parameters of a function of formula I by fitting the experimental data of the inert change of the physicochemical parameter of the at least second use,

c) determining the difference between the experimental data of the inert change of the physicochemical parameter of the at least second use and the function of formula I with the parameters determined in step b),

d) calculating the difference between the maximum value and the minimum value of the difference determined in step c) and normalizing said difference,

e) determining reduced separation efficacy of said re-useable chromatography column packing when the absolute value of the difference calculated in step d) is more than 0.1,

wherein the function of formula I is

2. The method according to claim 1, wherein said inert change is recorded during the purification by a standard conductivity measuring device.

3. The method according to claim 1 wherein said inert change is recorded during the purification by a standard adsorption measuring device.

4. The method according to claim 1, wherein said inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing is a change effected by the change of the concentration of a substance in the mobile phase that does not interact with the re-useable column packing.

5. The method according to claim 1, wherein said inert change is a change in conductivity, as measured by a standard conductivity measuring device.

6. The method according to claim 1, wherein said inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing is a change of from 100% of a solution containing a denaturing agent to 100% of a solution not containing said denaturing agent, or vice versa.

7. The method according to claim 6, wherein said denaturing agent is selected from sodium hydroxide, guanidinium chloride, urea or organic solvent.

8. The method according to claim 1, wherein said inert change is a sigmoid change.

9. The method according to claim 1, wherein said inert change is a change over time.

10. A method for the chromatographic purification of a polypeptide, wherein at least one chromatography step using a re-useable chromatography column packing is contained, wherein said method comprises the following steps:

a) identifying and determining the experimental data of an inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing of the at least second use,

b) determining the parameters of a function of formula I by fitting the experimental data of the inert change of the physicochemical parameter of the at least second use,

c) determining the difference between the experimental data of the inert change of the physicochemical parameter of the at least second use and the function of formula I with the parameters determined in step b),

d) calculating the difference between the maximum value and the minimum value of the difference determined in step c) and normalizing said difference,

wherein the function of formula I is

11. The method according to claim 10, wherein said inert change is recorded during the purification by a standard conductivity measuring device.

12. The method according to claim 10, wherein said inert change is recorded during the purification by a standard adsorption measuring device.

13. The method according to claim 10, wherein said inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing is a change effected by the change of the concentration of a substance in the mobile phase that does not interact with the re-useable column packing.

14. The method according to claim 10, wherein said inert change is a change in conductivity, as measured by a standard conductivity measuring device.

15. The method according to claim 10, wherein said inert change of at least one physicochemical parameter of a mobile phase passing through said re-useable chromatography column packing is a change of from 100% of a solution containing a denaturing agent to 100% of a solution not containing said denaturing agent, or vice versa.

16. The method according to claim 15, wherein said denaturing agent is selected from sodium hydroxide, guanidinium chloride, urea or organic solvent.

17. The method according to claim 10, wherein said inert change is a sigmoid change.

18. The method according to claim 10, wherein said inert change is a change over time.