Countercurrent Tangential Chromatography for Scalable Protein Purification
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Solution Overview
Problem
The biotechnology industry faces challenges in scaling up chromatography processes due to the limitations of column chromatography, including smaller column sizes, longer processing times, increased resin costs, and the inability to implement completely disposable downstream processes, which hinder the efficiency and cost-effectiveness of monoclonal antibody and protein purification.
Innovation Solution
A scalable and disposable chromatography system utilizing interconnected tangential flow filters and static mixers with countercurrent flow, where chromatography resin flows in a single pass, and buffers are pumped in a countercurrent direction to create concentration gradients, reducing buffer volume and increasing efficiency, while the permeate solution from the elution operation is collected as the purified product stream.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If column chromatography is used with larger column sizes to increase processing scale, then the scale of operation increases, but the columns cannot exceed 2 meter diameter due to physical limitations and the processing time increases to up to 24 hours
Solution Approach 1:
The invention divides the single large column into multiple smaller columns (first column, second column, third column) that operate in parallel. Each column processes a portion of the feed stream simultaneously, achieving large-scale processing without the physical limitations of a single oversized column. The columns are connected through a manifold system that distributes and collects streams efficiently.
Solution Approach 2:
The invention implements continuous operation where feed stream, wash stream, and elution stream are processed continuously through multiple columns in parallel. The system maintains continuous flow and processing without batch interruptions, eliminating the 24-hour cycle time of traditional batch column chromatography while maintaining large processing capacity.
2Reliability
If traditional column chromatography is used, then resin can be reused for multiple cycles, but the system cannot be completely disposable and requires cleaning validation
Solution Approach 1:
The invention enables complete disposability of the chromatography system by using multiple smaller columns that can be discarded after a single use. The parallel configuration and manifold design allow the entire system to be replaced rather than cleaned and validated, eliminating cleaning validation requirements and labor-intensive maintenance while maintaining reliable purification performance.
3Quantity of substance
If larger columns are used to process higher loads from improved upstream technology, then the load capacity increases, but the number of cycles required increases to at least 6 full cycles for a 20,000 L bioreactor
Solution Approach 1:
The invention segments the total load processing across multiple parallel columns, allowing simultaneous processing of different portions of the feed stream. This parallel architecture enables the system to handle large loads from 20,000 L bioreactors in a single continuous pass rather than requiring 6 sequential cycles, dramatically improving productivity while maintaining high load capacity.
Solution Approach 2:
The manifold system provides multi-functionality by serving as both a distribution system for feed stream and a collection system for purified product. The same manifold infrastructure supports multiple columns operating in parallel, enabling the system to process large loads efficiently without requiring separate processing trains for different operational modes.
4Productivity
If membrane chromatography is adopted as an alternative to column chromatography, then some improvements are achieved, but it still cannot be completely disposable and requires significant capital equipment investment
Solution Approach 1:
The invention uses simple, disposable chromatography columns connected through a manifold system rather than expensive, complex membrane chromatography equipment. The columns can be discarded after single use, eliminating the need for expensive membrane modules and associated cleaning/validation infrastructure, significantly reducing capital equipment investment while maintaining high process efficiency through parallel operation.
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 enables larger-scale operation, faster processing, reduced resin costs, and the potential for completely disposable systems, significantly enhancing the efficiency and cost-effectiveness of protein purification processes.
Implementation Method 1
The tangential flow filters separate the purified product stream from the resin slurry
Implementation Method 2
permeate solutions from later stages are recycled back into previous stages
Implementation Method 3
This creates concentration gradients in the permeate solutions of the tangential flow filters in the countercurrent direction to resin flow
Implementation Method 4
buffers are pumped into the module in a countercurrent direction to the flow of resin
Implementation Method 5
The static mixers enhance mixing between the resin slurry and buffer solutions
Data Source
Figure 1
Figure 2A~2B
Figure 3
AI summary
This invention relates to a breakthrough in the art of chromatography allowing 1) larger scale of operation; 2) faster processing time; 3) disposability; 4) reduction of media/resin expenses; and 5) a reduction of capital equipment investment. In this invention, the chromatography column is replaced by a module that consists of two or more interconnected tangential flow filters and static mixers. The chromatography resin flows through this module in a single pass, while similar operations to a regular chromatographic process are performed on the resin (binding, washing, elution, regeneration, and equilibration). The buffers for these operations are pumped into the module in a countercurrent direction to the flow of resin, and permeate solutions from later stages are recycled back into previous stages. This creates concentration gradients in the permeate solutions of the tangential flow filters in the countercurrent direction to resin flow, thus saving buffer volume and increasing process efficiency.