EP-4736979-A1 - CHROMATOGRAPHY COLUMN PACKING MEDIUM RECOVERY

EP4736979A1EP 4736979 A1EP4736979 A1EP 4736979A1EP-4736979-A1

Abstract

Systems include a chromatography column tube having an inlet and outlet and port assembly arranged in a wall of the column tube between flow distributors that together form a chamber within the tube that is filled with packing medium are described. The port assembly facilitates the removal of resin from the pre-packed column and the port assembly does not affect fluid flow in the normal use of the column for chromatographic separation or ability to maintain sanitary conditions within the column. Also described are methods that include attaching tubing to a pump and to a column inlet and a column outlet, opening the port assembly, attaching tubing to the port assembly and to a second reservoir, and pumping aqueous solution from a reservoir into the chamber and out through the port assembly into the second reservoir, thereby removing packing medium from the column along with the flowing aqueous solution.

Inventors

WARD, Travis R.
PEYSER, JAMES RONALD
PENTIA, Dana
FRANKEL, Evan

Assignees

Repligen Corporation

Dates

Publication Date: 20260506
Application Date: 20170612

Claims (12)

A method of recovering packing medium from a chamber between flow distributors within a pre-packed chromatography column tube having an inlet and an outlet and a port assembly arranged in a wall of the column tube between first and second flow distributors, the method comprising: accessing an end of a plug extending outside a port of the port assembly, and removing the plug from the port; attaching tubing between the port assembly and a recovery tank; and collecting, in the recovery tank, packing medium from within the chromatography column.
The method of claim 1, wherein removing the plug from the port further comprises removing the plug from a port extending through an opening in the chromatography column tube wall; optionally wherein removing the plug from the port further comprises removing a sanitary clamp from the port assembly to release the plug from the port assembly.
The method of claim 1 or 2, further comprising: attaching tubing between a reservoir tank and the inlet and the outlet; and pumping solution from within the reservoir tank and into the chamber within the chromatography column through the inlet and outlet, optionally wherein the recovery tank is the reservoir tank or the recovery tank is separate from the reservoir tank.
The method of any of claims 1-3, further comprising pumping packing medium out of the chromatography column tube through the port assembly and into the recovery tank; pumping packing medium containing solution out of the chromatography column tube through the port assembly and into the recovery tank, thereby removing packing medium from the column along with the solution; and/or filtering and collecting the packing medium through a mesh in the recovery tank.
The method of any one of claims 1-4, further comprising pumping a gas into the chromatography column chamber in an amount and at a volume sufficient to suspend the packing medium in the column; pumping a gas into the chromatography column chamber and out through the port assembly and into the recovery tank, thereby removing packing medium from the column; pumping the solution from the reservoir tank to fill the chromatography column chamber and suspend any remaining packing medium particles therein; and/or pumping gas and the solution to fill the chromatography column chamber until a desired amount of the packing medium is pumped into the recovery tank.
A chromatography column comprising: a column tube comprising a cylindrical wall having a first end and a second end; a first flow distributor arranged within a first end of the column tube, and a second flow distributor arranged within a second end of the column tube, the first flow distributor and the second flow distributor defining a chamber therebetween; and a port assembly extending through an opening in a wall of the column tube and in fluid communication with the chamber; wherein the port assembly comprises a port configured to permit removal of a medium therethrough from within the chamber, and a plug removably extending through the port and having an end sealed against an end of the port extending away from the opening in the column tube wall.
The chromatography column of claim 6, wherein: the port is removably extended through the opening in the column tube wall; the end of the plug sealed against the end of the port is sealed with a gasket and secured within the port assembly by a clamp, and is configured to facilitate removal of the plug from the port.
The chromatography column of claim 6 or 7, wherein the port is configured to be secured within the opening in the column tube wall.
The chromatography column of any one of claims 6-8, wherein the port comprises a beveled flange within the inside surface of the column tube wall.
The chromatography column of claim 9, wherein the flange of the port projects less than 6 mm past an inside surface of the column tube wall.
The chromatography column of claim 9, wherein the flange of the port is configured to avoid zones without fluid accessibility in a fluid inside the column.
The chromatography column of any one of claims 6-11, wherein the chromatography is prepacked with a packing medium contained within the chamber, the packing medium being packed into the column through one of the first and second ends thereof, and removed from the column via the opening in the wall of the column.

Description

CROSS-REFERENCE TO RELATED APPLICATION This application claims priority to U.S. Patent Application 62/348,760, filed on June 10, 2016, entitled: "Chromatography Column Packing Medium Recovery,". TECHNICAL FIELD This invention relates to methods and system for removing and recovering column packing medium from chromatograph columns. BACKGROUND Column chromatography is a separation and/or purification technique in which a stationary "bed" of a packing medium is contained within a rigid tube. The packing medium can be in the form of particles of a solid ("stationary phase") or a solid support material coated with a liquid stationary phase. Either way, the packing medium typically fills the inside volume of the column tube. In separation chromatography, as a liquid sample ("mobile phase") passes through the column, different compounds in the sample can associate differentially with the stationary phase (e.g., packing medium) such that they are slowed relative to the mobile phase and move through the column at different speeds. Thus, those compounds that associate more with the stationary phase move more slowly through the column than those that associate less, and this speed differential results in the compounds being separated from one another as they pass through and exit the column. Features of the stationary phase that promote differential association can be, e.g., ionic charge (ion exchange chromatography), hydrophobicity (hydrophobic interaction chromatography), and porosity (size exclusion chromatography). In yet another type of column chromatography, affinity chromatography, the packing medium includes binding agents, such as antigens, antibodies, or ligands, that specifically bind to one or more desired compounds or molecules in the liquid sample. Thus, as the liquid sample flows through the packing medium only the desired compounds or molecules remain in the column. A subsequent flow through the packing medium of an eluting liquid separates the desired compounds or molecules from the binding agents attached to the packing medium, or separates the binding agents from the packing medium. Either way, the desired compounds or molecules are rinsed out of the column and collected in the eluting fluid. Affinity chromatography can be used in a number of applications, including nucleic acid purification, protein purification from cell free extracts, and purification from blood. The main components of a chromatography column are the column or tube, which is often made of a metal, glass, or highly rigid plastic material, and a pair of flow distributors, which are typically inserted into the two ends of the tube to form a space or chamber in the tube between the flow distributors into which the packing medium is loaded. Chromatography columns packed with separation media on site usually have an adjustable top distributor that allows for adjusting the top flow adaptor position or removal of the adaptor to recover the separation media for regeneration or reprocessing in the event that the column has lost performance. Performance can be lost due to process deviations such as inadvertently pumping the solution-filled separation media dry or through repeated process cycles and slow deterioration of performance due to feed stream fouling of the separation media. Chromatography columns can be pre-packed and used upon demand for development and commercial bioprocess manufacturing. These columns are produced and fixed in bed height such that flow distributors are irreversibly aligned within the chromatography tube. While pre-packed columns have the advantage over conventional on site packed columns of ease of use and overall economic advantage, the inability to recover and reprocess the separation media is a disadvantage. SUMMARY The invention is based, at least in part, on the discovery that if you provide a packing medium recovery port on a chromatography column arranged between the two flow distributors, you can then simply and quickly remove the packing medium in the chromatography column. In addition, by using a specific design of the packing mediumrecovery port, you can avoid disruption of the chromatography flow path and the formation of dead zones within the column. The present discovery includes the recognition that recovering media from a prepacked column that has lost performance must be done in a way that does not damage or contaminate the separation media. Any port included in a pre-packed column must not interfere with the chromatographic separation properties or the ability to maintain a sanitary condition of the column when used repeatedly. The present systems and methods provide pre-packed chromatography columns that allow recovery of the separation media in an intact and unadulterated form and that do not impair the separation performance of the column or increase the risk of increasing an unwanted bioburden during normal use of the column. Accordingly, the present disclosure describes a series of chroma