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EP-3697515-B1 - METHODS FOR CHROMATOGRAPHY RESIN SLURRY DETERMINATION

EP3697515B1EP 3697515 B1EP3697515 B1EP 3697515B1EP-3697515-B1

Inventors

  • MAO, Nathan
  • SHIERLY, ERIC
  • GALLEA, JOSEPH
  • FENG, RAYMOND
  • DISSANAYAKE, TIKIRI
  • SCHILLING, BERNHARD
  • CARVER, Scott

Dates

Publication Date
20260513
Application Date
20181015

Claims (14)

  1. A method of determining a slurry concentration, comprising: adding a slurry comprising a resin to a first chromatography column; pumping a volume of liquid through the first chromatography column at a constant flow rate; controlling the flow rate to match a consolidation rate employed by a second chromatography column to form a consolidated resin in the first chromatography column, wherein the volume of liquid is pumped through the first chromatography column by an automated pump system, wherein the automated pump system is programed to cycle on and off for a predetermined period of time or for multiple periods of time; upon completion of forming the consolidated resin, measuring a bed height of the consolidated resin in the first chromatography column; and determining a slurry concentration to pack the second chromatography column, wherein the packed second chromatography column has a packing factor within 5% of an ideal packing factor.
  2. The method of claim 1, wherein the liquid is deionized water, distilled water, purified water, a saline solution, or an organic solvent.
  3. The method of claim 1, wherein the step of adding the slurry further comprises adding water to the first chromatography column and allowing the slurry to settle for about five minutes to about one hour prior to pumping.
  4. The method of claim 1, wherein the second chromatography column comprises a consolidation rate and wherein the flow rate of the first chromatography column is equivalent to the consolidation rate of the second chromatography column.
  5. The method of claim 1, wherein the automated pump system comprises a syringe operatively coupled to a pump.
  6. The method of claim 1, further comprising, pausing the automated pump to stop the flow of liquid through the first chromatography column and allowing the resin to settle, and restarting the pump to flow liquid through the first chromatography column before completion of the pumping step.
  7. The method of claim 6, wherein the pumping is paused for more than one time period.
  8. The method of claims 6 or 7, wherein the pumping is paused for a time period of 1 to 60 minutes.
  9. The method of claim 1, wherein the step of setting the automated pump further comprises providing a flow rate based on which type of resin is utilized in the slurry and the consolidation rate employed by the second chromatography column.
  10. The method of claim 9, wherein the flow rate ranges from about 20 cm/hr to about 1500 cm/hr.
  11. The method of claim 1, wherein: (a) the method further comprises mixing the slurry prior to adding the slurry to the chromatography column; and (b) the liquid is deionized water, distilled water, purified water, a saline solution, or an organic solvent.
  12. The method of claim 1, wherein: (a) the method further comprises mixing the slurry to uniformly distribute the resin prior to adding the slurry to the chromatography column; (b) the automated pump system comprises a pump having a controller, wherein the automated pump system is configured to pump a volume of water through the first chromatography column and the controller is configured to control a flow rate of the volume of water to match a consolidation rate employed by a second chromatography column that will pack the resin; and (c) the method further comprises the step of inputting the determined slurry concentration into the automated pump system after the step of determining the slurry concentration to pack the second chromatography column.
  13. The method of claim 12, wherein: (a) the flow rate ranges from about 30 cm/hr to about 60 cm/hr; and (b) the second chromatography column comprises a consolidation rate and the flow rate employed by the first chromatography column is equivalent to the consolidation rate of the second chromatography column.
  14. The method of any one of the preceding claims, where the packed second chromatography column has a bed height within 1 cm of a target bed height.

Description

FIELD OF THE INVENTION The present invention relates to methods for resin slurry determination for a slurry of a chromatography resin for use during a column packing. More specifically, the present invention provides a more accurate and consistent method for determining a slurry concentration for packing a chromatography column, for example by mode of axial compression. BACKGROUND OF THE INVENTION Resin slurry determination is a method commonly used to determine the amount of chromatography media present in aqueous-based slurries. The resin slurry percentage or slurry concentration is used in calculations during chromatography column packing operations to determine how much aqueous slurry should be put in a column to ensure proper operation and target bed heights. The slurry concentration is the major variable that must be determined and input into automatic packing columns, for example, AxiChrom® columns, to consistently pack to the correct height and packing factor. Indeed, automatic packing columns, such as AxiChrom® columns, provide packing operations that are highly automated, but rely on an accurate slurry concentration to perform reproducibly. Methods for automatic methods for column packing are known from WO2008/109192 or WO2009/145715. Current methods of resin slurry determination include gravity settling, centrifugation, and flow consolidation. The current methods obtain a representative sample from the resin slurry bulk and submit the sample to a force that induces resin settling which in turn allows for analysis of the ratio of settled resin to supernatant (the slurry concentration). However, there are several disadvantages with these methods. For instance, in the current methods, discrepancies between slurry concentration results and achieved Packing Factor (defined as the ratio of consolidated bed height to final packed bed height) and column height are often observed. In addition, centrifugation results have been shown to have high variability between different resin types and different ratios of the same standard if various wait times are employed after centrifugation. Gravity settling is considered more consistent, but can take considerable time to complete and may require a correction factor to be empirically determined and applied which increases method uncertainty. Recently, GE Healthcare has provided a recommended consolidation method specifically for the automated packing column, AxiChrom®. This method involves obtaining a representative sample from the resin bulk, loading it into a 10 mm diameter by 100 mm Tricorn column, and manually operating a syringe to create consolidation force on the sample. After final consolidation is achieved, resin slurry is determined from the ratio of resin to supernatant. However, this method is inconsistent largely due to variation in how hard the plunger on the syringe is depressed. Accordingly, there remains a need for a more accurate and consistent method for determining a slurry concentration for packing a chromatography column, for example, automated packing chromatography columns. SUMMARY OF THE INVENTION Methods are provided for determining a resin slurry percentage or slurry concentration for a slurry of a chromatography column. The methods of the present invention provide highly accurate slurry concentrations that in turn provide for accurate target bed heights when packing a resin in a chromatography column, for example, in an automated packing column. The present invention is defined in the appended claims. Also disclosed, but not part of the present invention, is a method of determining a slurry concentration, including adding a slurry comprising a resin to a first chromatography column; pumping a volume of liquid, for example, deionized water, distilled water, purified water, a saline solution, or an organic solvent, through the chromatography column at a constant flow rate to form a consolidated resin, where the volume of liquid is pumped through the chromatography column by an automated pump system; upon completion of the pumping step, measuring a bed height of the consolidated resin; and determining a slurry concentration from the measured bed height. In some embodiments, the method may further include a step of adding water to the chromatography column and allowing the slurry to settle for about five minutes to about one hour prior to pumping. In other embodiments, the method may further include utilizing the determined slurry concentration to pack a second chromatography column. In this aspect, the second chromatography column includes a consolidation rate and the flow rate is equivalent to the consolidation rate. In still another embodiment, the automated pump system may include a syringe operatively coupled to a pump. In some embodiments the pump system is programed to cycle on and off for a predetermined period of time or for multiple periods of time. Also disclosed, but not part of the present invention, is a method for determining