Perfusion Module With Pressure-Regulating Airflow for Low Shear Culture
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Solution Overview
Problem
Current perfusion culture systems face challenges in scaling up to large volumes (1000-30,000 L) due to high costs and inefficiencies in cell retention, particularly with existing alternating tangential flow systems, which are costly and have dead volumes that affect cell culture quality.
Innovation Solution
A perfusion culture system comprising a bioreaction module, a perfusion module with pressure-regulating airflow, and a filtration module, where each perfusion chamber is connected to a pressure-regulating pipe with a pump for airflow control, allowing for positive pressure control and reducing shear forces, and multiple chambers work alternately to achieve large-scale perfusion culture.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If filtration-based cell retention devices are used to achieve complete cell isolation, then cell isolation rate is improved to 100%, but filter membranes are easily clogged by cell debris and defoamers, leading to termination of perfusion culture
Solution Approach 1:
The invention extracts and removes harmful substances (cell debris, defoamers, and other contaminants) from the culture medium before the medium reaches the filter membrane. By placing a pre-filtration system or debris removal device upstream of the main filter, these contaminants are separated and removed, preventing them from clogging the filter membrane while maintaining complete cell isolation through the main filter.
Solution Approach 2:
The invention introduces an intermediary component (such as a pre-filter, skimmer, or debris removal device) between the culture medium and the main filter membrane. This intermediary captures and removes contaminants before they can reach and clog the main filter, acting as a protective barrier that extends the operational life of the filter while maintaining its cell isolation function.
2Object-affected harmful factors
If centrifugation is used for cell isolation, then cell damage is avoided, but cell isolation completeness is reduced and cell isolation effect is affected by perfusion rates
Solution Approach 1:
The invention replaces the centrifugal separation mechanism with a filtration-based system. Instead of using centrifugal force that may damage cells, the system uses a filter membrane with appropriate pore size to physically separate cells from the culture medium. This mechanical substitution eliminates cell damage while achieving complete cell isolation, as the filter membrane can retain all cells regardless of perfusion rate variations.
Solution Approach 2:
The invention employs a filter membrane with specifically designed pore size and structure to achieve complete cell retention. The porous material is selected to have pores small enough to retain all cells in the culture while allowing culture medium to pass through freely. This ensures 100% cell isolation completeness without causing cell damage, overcoming the limitations of centrifugation-based methods.
3Object-affected harmful factors
If sedimentation is used for cell isolation, then cell damage is avoided, but cell isolation completeness is reduced and cell isolation effect is affected by perfusion rates
Solution Approach 1:
The invention replaces the sedimentation process with a filtration system. Instead of relying on gravitational settling that is affected by perfusion rates and may not achieve complete separation, the system uses a filter membrane to physically block cells from passing through. This substitution ensures complete cell isolation (100% retention) while avoiding cell damage, as the filtration process is not influenced by flow rate variations in the same way sedimentation is.
Solution Approach 2:
The invention uses a filter membrane with optimized pore size to achieve complete cell retention. The porous structure of the filter is designed with pores smaller than the smallest cells in the culture, ensuring that no cells can pass through regardless of perfusion rate. This provides consistent, complete cell isolation without the limitations of sedimentation-based methods.
4Reliability
If alternating tangential flow systems are used for cell retention, then membrane clogging is relieved and shear force is reduced, but system cost increases and dead volume affects cell culture quality
Solution Approach 1:
The invention inverts the traditional approach by using a simple filtration system without complex flow reversal mechanisms. Instead of using alternating tangential flow to prevent clogging, the system uses a filter design that inherently resists clogging through proper pore size selection and placement of debris removal devices upstream. This simpler approach achieves comparable or better clogging resistance without the complexity and cost of ATF systems.
Solution Approach 2:
The invention extracts and removes contaminants (cell debris, defoamers, and other particulates) from the culture medium before they can reach and clog the filter membrane. By placing debris removal devices or pre-filtration systems upstream, the load on the main filter is significantly reduced, preventing clogging without requiring complex flow reversal mechanisms or high shear force alternating flows.
5Productivity
If scale-up of perfusion culture is achieved using conventional systems, then production volume increases, but system cost increases and operational flexibility decreases
Solution Approach 1:
The invention segments the perfusion culture system into modular units that can be easily scaled by adding or removing modules. Each module contains a filter, perfusion chamber, and associated components as separate, interchangeable units. This modular segmentation allows production volume to be increased by simply adding more modules in parallel or series, without proportionally increasing system complexity or cost, thereby maintaining operational flexibility while achieving scale-up.
Solution Approach 2:
The invention designs universal, multi-functional components that can be used across different scale configurations. The modular modules are designed to be identical or compatible across all scale levels, allowing the same basic unit to serve multiple functions and be deployed in various configurations. This universality reduces development and manufacturing costs while enabling flexible scaling from small to large production volumes.
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
The system enables efficient large-scale perfusion culture (1000-30,000 L) while minimizing shear forces and operational costs, facilitating high cell density and protein yield with improved scalability and flexibility.
Implementation Method 1
designed based filtration, sedimentation, and centrifugation
Implementation Method 2
designed based filtration, sedimentation, and centrifugation
Implementation Method 3
designed based filtration, sedimentation, and centrifugation
Implementation Method 4
each perfusion chamber is connected to a pressure regulating pipe for inflow or outflow of an airflow, and the pressure regulating pipe is provided with a pump that enables the pressure regulating pipe to be opened and closed and changes a delivery direction of the airflow
Implementation Method 5
effectively relieve membrane clogging, a low shear force
Implementation Method 6
achieves sufficient flushing of the hollow fibers under low shear forces, and effectively reduces clogging caused by filter cakes accumulated on the hollow fibers
Data Source
AI summary
A perfusion module and a perfusion culture system are provided, including: a bioreaction module, including at least one bioreactor; a perfusion module, including at least one perfusion chamber, each perfusion chamber is connected to a pressure regulating pipe for inflow or outflow of an airflow; when there are multiple perfusion chambers, every two adjacent perfusion chambers are connected to each other; at least one bioreactor, the perfusion module, the filtration module are communicated in sequence, if there are more than two perfusion chambers, all of perfusion chambers are set side by side; a cell culture fluid in at least one bioreactor flows into at least one perfusion chamber, flows out from at least one perfusion chamber into at least one filter, and a permeate from at least one filter flows into a harvesting device. The present disclosure avoids high shear forces in the perfusion culture system.