Overload Ion Exchange Chromatography Column Regeneration
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Solution Overview
Problem
Overload cation-exchange chromatography leads to column fouling during sanitization due to high load density, resulting in increased pressure and impurity carry-over, which is not effectively minimized in existing methods.
Innovation Solution
A method involving the use of regeneration buffers containing sodium acetate or Tris and sodium acetate, followed by sanitization with 0.5 N NaOH, and storage in 0.1 N NaOH buffer to clean and reuse ion exchange chromatography materials, particularly in overload mode for polypeptide purification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If overload chromatography is used to intensify the process and improve resin utilization, then productivity is improved, but column fouling occurs during sanitization leading to increased pressure
Solution Approach 1:
The patent applies preliminary action by performing a regeneration step before sanitization. The regeneration buffer (comprising sodium acetate and/or Tris and sodium acetate) is passed through the column to remove bound host-cell protein and impurities before the sanitization phase. This preliminary cleaning action prevents column fouling during subsequent sanitization, thereby preventing pressure increases while maintaining the high productivity benefits of overload chromatography.
2Productivity
If overload chromatography is used to improve resin utilization, then productivity is improved, but impurity carry-over occurs between runs
Solution Approach 1:
The regeneration buffer is passed through the column before sanitization to perform preliminary removal of bound impurities and host-cell protein. This ensures the column is properly cleaned and prepared, preventing impurity carry-over to the next chromatography run while maintaining high resin utilization during the overload process.
Solution Approach 2:
The regeneration buffer acts as an intermediary cleaning step between the overload chromatography run and the sanitization phase. It serves as a mediator that removes bound impurities and prepares the column for effective sanitization, thereby preventing impurity carry-over while allowing the column to be reused with high productivity.
3Productivity
If high load density is used in overload mode, then purification efficiency is improved, but column fouling increases during sanitization
Solution Approach 1:
The regeneration buffer is passed through the column before sanitization to perform preliminary removal of bound host-cell protein and impurities. This preliminary action prevents column fouling during the subsequent sanitization phase, allowing the column to maintain its performance and be reused effectively while continuing to operate at high load density for efficient purification.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach effectively reduces pressure increases during sanitization and prevents impurity carry-over, enabling the reuse of ion exchange chromatography materials while maintaining purification efficiency.
Implementation Method 1
Overload (OL) cation-exchange chromatography has been shown to have equivalent impurity removal capabilities to bind and clutch mode
Implementation Method 2
the column fouls (high column pressure, >80 psi) during the sanitization phase (0.5N Sodium Hydroxide)
Implementation Method 3
storing the ion exchange chromatography material in a storage buffer, wherein the storage buffer comprises about 0.1 N NaOH
Data Source
AI summary
The present invention provides methods for cleaning or regenerating a chromatography material for reuse. The methods of the invention can be used for cleaning or regenerating ion exchange chromatography columns for reuse in the large-scale manufacture of multiple polypeptide products where the ion exchange chromatography column in used in overload mode.