Direct capture using large bead chromatography media

Liquid chromatography column designed to be radially flown through by the process liquid and comprising a packed bed of beads of between approximately 120 micrometer and approximately 1 millimeter diameter and these beads are hydrophilic, which beads are designed to capture biologics from mammalian CHO cells or cell culture or cell fermentation harvests having a diameter of 0.1-10% of bead diameter from the process liquid, the packed bed is held between an outer frit and an inner frit of the column and the packed bed is torus shaped, wherein the following is applied:

• the outer frit has a first surface area and the inner frit has a second surface area which is smaller than the first surface area;
• the ratio between the first surface area and the second surface area is equal to the inlet-frit surface area/outlet-frit surface area (I/O) ratio of the column and is less than 5:1 such that the first surface area is not more than 5 times the second surface area;
• the outer frit and the inner frit have an hydrophilic surface and each the outer frit and the inner frit comprising at least two directly on the top of each other laid layers or sheets of woven wires of stainless steel, or, the outer frit and the inner frit being made from stainless steel and have a thickness of at least 0.3 mm, the outer frit being adjacent an axially extending external housing wall of the column, the inner frit being disposed closer to a central longitudinal axis of the column, a core member being disposed in the space delimited by the inner frit, the core member having an external wall delimiting an inner flow channel with the inner frit, the inner flow channel having a width of at least 0.5 mm, the inner flow channel tapering in an axial direction from one end to an opposite end of the column along the core member;
• the column being designed such that the liquid to be processed flowing to first being distributed across the surface of the outer frit, subsequently passing through the outer frit, subsequently flowing radially inward through the packed bed to arrive at the inner frit, subsequently passing through the inner frit to enter into the inner flow channel and subsequently passing in axial direction along the core member and finally exiting the column.

Set of mutually identical liquid chromatography columns each column of the set designed to be radially flown through by the process liquid and each column of the set comprising a packed bed of beads of between approximately 120 micrometer and approximately 1 millimeter diameter and these beads are hydrophilic, which beads are designed to capture biologics from mammalian CHO cells or cell culture or cell fermentation harvests having a diameter of 0.1-10% of bead diameter from the process liquid, the packed bed is held between an outer frit and an inner frit of the column and the packed bed is torus shaped, wherein the following is applied for each column of the set:

• the outer frit has a first surface area and the inner frit has a second surface area which is smaller than the first surface area;
• the ratio between the first surface area and the second surface area is equal to the inlet-frit surface area/outlet-frit surface area (I/O) ratio of the column and is less than 5:1 such that the first surface area is not more than 5 times the second surface area;
• the outer frit being adjacent an axially extending external housing wall of the column, the inner frit being disposed closer to a central longitudinal axis of the column, a core member being disposed in the space delimited by the inner frit, the core member having an external wall delimiting an inner flow channel with the inner frit, the inner flow channel having a width of at least 0.5 mm, the inner flow channel tapering in an axial direction from one end to an opposite end of the column along the core member;
• each column of the set being designed such that the liquid to be processed flowing to first being distributed across the surface of the outer frit, subsequently passing through the outer frit, subsequently flowing radially inward through the packed bed to arrive at the inner frit, subsequently passing through the inner frit to enter into the inner flow channel and subsequently passing in axial direction along the core member and finally exiting the column
• wherein the set of columns is designed such that the process liquid is fed to a subset of at least two of the set of identical columns directly from the source without any intermediate filtration or other clarification equipment, nor any harvest holding-step or holding vessel and the process liquid, depleted by the subset of columns from the product of interest, is leaving said subset of columns after having flown radially through each of the columns of the subset of columns,
• the columns of the subset of at least two of the set of identical columns are connected in series with each other and the subset is designed such that the process liquid is supplied to the most upstream column of the subset, flows successively through the in series connected columns and leaves the subset through the most downstream column and after time elapse, if the most upstream column is become saturated with product, the most upstream column is disconnected from the subset and removed from the series connection and a replacement column from the set is added to the subset such that it is connected in series downstream from the most downstream column of the subset and this process is repeated and the column removed from the subset is off-line processed to reset the packed bed of this column such that this column is made ready to become available for the subset to become saturated again by connecting this column in series as the most downstream column of the subset.