Tandem AEX-CEX Chromatography Without In-Line Buffer Adjustment
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Solution Overview
Problem
Current downstream protein purification processes, particularly for monoclonal antibodies and their derivatives, are inefficient, costly, and time-consuming due to the need for in-line adjustments of pH and conductivity in chromatography steps, leading to increased operation costs and skid footprints.
Innovation Solution
A combined chromatography process using cation exchange chromatography (CEX) and anion exchange chromatography (AEX) in tandem, operated in flow-through mode without adjustments to pH or conductivity, processing the columns as a single unit to reduce process costs, time, and skid footprint.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Extent of automation
If in-line adjustments of buffer conditions (pH and conductivity) are performed between chromatography steps, then the purification process can be automated and controlled, but the operation time and skid footprint increase
Solution Approach 1:
The patent combines multiple chromatography steps (AEX and CEX) into a single integrated unit operation where the effluent from one column directly feeds into the next column without intermediate buffer adjustments. This merging of steps eliminates the time required for in-line buffer adjustments while maintaining automation capability.
Solution Approach 2:
The patent implements continuous flow-through mode operation where the chromatography columns are connected in series and operated continuously without interruption for buffer adjustments. The loading buffer flows continuously through both columns, and the chase buffer is applied continuously, eliminating idle time between operations.
2Manufacturing precision
If in-line adjustments of buffer conditions are performed for each unit operation, then the chromatography process can be optimized for each step, but the skid footprint and operation cost increase
Solution Approach 1:
The patent merges the AEX and CEX chromatography steps into a single integrated unit operation with direct column connection, eliminating the need for separate buffer adjustment equipment and reducing the overall skid footprint while maintaining optimized separation conditions.
Solution Approach 2:
The loading buffer and chase buffer serve multiple functions across both chromatography steps, reducing the number of separate buffer systems needed and simplifying the overall equipment configuration required for the purification process.
3Ease of manufacture
If traditional batch purification processes are used with hold tanks between unit operations, then buffer adjustments can be made for each step, but the process efficiency and productivity decrease
Solution Approach 1:
The patent transitions from batch processing with hold tanks to continuous flow-through mode operation, where the chromatography columns are operated continuously without interruption. This continuous operation eliminates the inefficiencies of batch processing while maintaining buffer adjustment flexibility through the loading and chase buffer system.
Solution Approach 2:
The patent removes the hold tank intermediate steps from the purification process, directly connecting the chromatography columns in series. This extraction of intermediate holding steps eliminates the time loss associated with batch processing while preserving the ability to make necessary buffer adjustments.
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 contaminants such as host cell proteins, DNA, and residual protein A, achieving high throughput and minimizing buffer transition peaks, thereby enhancing the purification efficiency and reducing operational expenses.
Implementation Method 1
The chromatography medium of the second chromatography column is different from the chromatography medium of the first chromatography column. In one embodiment, the first chromatography column is a cation exchange chromatography column and the second chromatography column is an anion exchange chromatography column.
Implementation Method 2
In one embodiment, the first chromatography column is a cation exchange chromatography column and the second chromatography column is an anion exchange chromatography column.
Implementation Method 3
Methods of purifying proteins using chromatography
Data Source
AI summary
This disclosure provides a method of purifying proteins using a combined AEX-CEX chromatography in flow-through mode under an operating condition wherein no automation, engineering control, and/or in-line adjustment is used between the AEX and the CEX. The AEX and CEX (or CEX and AEX) can be connected directly.