Method for Cleaning of Packed Bed Chromatography Columns

1 . A process for chromatographic separation of at least one target biomolecule from at least one contaminant, comprising the steps of:

a) providing an axial chromatography column comprising a consolidated bed of separation matrix particles, said consolidated bed being confined between a bottom support net and a movable top adaptor;

b) separating a target biomolecule from at least one contaminant on the column;

c) raising the adaptor by at least 10% of the height of the consolidated bed;

d) flowing a first cleaning liquid upwards through the bed under conditions sufficient to liquefy the bed, and;

e) repacking the matrix particles of the liquefied bed to create a consolidated bed and lowering the adaptor such that it contacts the packed bed and optionally compresses it, and;

f) separating a target biomolecule from at least one contaminant on the column.

2 . A process for chromatographic separation of at least one target biomolecule from at least one contaminant, comprising the steps of:

a) providing an axial chromatography column comprising a consolidated bed of separation matrix particles, said consolidated bed being confined between a bottom support net and a movable top adaptor;

b) separating a target biomolecule from at least one contaminant on the column;

c) unpacking said consolidated bed and conveying the matrix particles to a separate cleaning vessel;

d) contacting the matrix particles in the cleaning vessel with a first cleaning liquid;

e) conveying the cleaned matrix particles back to the column and repacking the consolidated bed, and;

f) separating a target biomolecule from at least one contaminant on the column.

3 . The process of claim 1 , further comprising after step b) a step b′) of cleaning the column by conveying a second cleaning liquid through the consolidated bed.

4 . The process of claim 1 , further comprising after step f) a step f′) of cleaning the column by conveying a third cleaning liquid through the consolidated bed.

5 . The process of claim 3 , further comprising after step f) a step f′) of cleaning the column by conveying a third cleaning liquid through the consolidated bed and further comprising repeating steps b) and b′) and/or steps f) and f′) at least once.

6 . The process according to claim 1 , further comprising a step of storing the column for at least one week between steps b) and f).

7 . The process according to claim 1 , further comprising agitating the liquefied bed in step d).

8 . The process according to claim 7 , wherein the agitation comprises vibrating or sonicating the liquefied bed or the matrix particles.

9 . The process according to claim 7 , wherein the agitation comprises agitation by jets from one or more nozzles.

10 . The process according to claim 7 , wherein the agitation comprises an oscillating flow in step d).

11 . The process according to claim 1 , wherein the separation matrix particles, when equilibrated with pure water, have a density of about 1.0-1.4 g/ml.

12 . The process according to claim 1 , wherein the matrix particles comprise immobilized proteinaceous ligands, such as Protein A, Protein G, Protein L, single chain camelid antibodies or functional varieties thereof.

13 . The process according to claim 1 , wherein the first cleaning liquid is alkaline, with a pH value of at least 10.

14 . The process according to claim 1 , wherein the inner diameter of the chromatography column is at least 2.5 cm.

15 . The process according to claim 1 , wherein the height of the consolidated bed is about 5-50 cm.

16 . The process according to claim 1 , wherein the ratio between the inner diameter of the chromatography column to the height of the consolidated bed is at least 0.4.

17 . The process according to claim 1 , wherein the matrix particles are essentially spherical, with average diameter at least 20 μm.

18 . The process according to claim 1 , wherein the coefficient of variation of the matrix particle size distribution is about 10-50%.

19 . The process according to claim 1 , wherein the matrix particles comprise a polysaccharide support.

20 . The process according to claim 1 , wherein the upward flow velocity in step d) is about 100-500 cm/h and the viscosity of the first cleaning liquid is about 1.0-1.5 mPas.

21 . The process according to claim 1 , further comprising, between steps d) and e), a step of removing a sample of the cleaned matrix particles and analyzing the sample with respect to purity.

22 . The process according to claim 1 , further comprising, after step f), a step of recovering the second target biomolecule and, optionally after further purification, using it in a pharmaceutical formulation.

23 . A method for cleaning and/or sanitization of a packed bed chromatography column, comprising the steps of:

a) providing an axial chromatography column comprising a consolidated bed of separation matrix particles, said consolidated bed being confined between a bottom support net and a movable top adaptor;

b) raising the adaptor by at least 10% of the height of the consolidated bed;

c) flowing a cleaning liquid upwards through the bed under conditions sufficient to liquefy the bed, and;

d) repacking the matrix particles of the liquefied bed by flowing a packing fluid downwards through the column to create a consolidated bed and lowering the adaptor such that it contacts the packed bed and optionally compresses it.