Phase-Change Hydrogel Cell Separation via Counter-Current Centrifugation
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Solution Overview
Problem
Current magnetic separation techniques for isolating target cells for immunotherapies are time-consuming, detrimental to cell viability, and require extensive processing to meet FDA standards, including the removal of magnetic particles, which can take several days and compromise cell health.
Innovation Solution
The method involves specific binding of target cells to phase-change hydrogel compositions, followed by counter-current centrifugation for separation, allowing for rapid and viable cell isolation without the need for magnetic particles or manual manipulation, using biological moieties like antibodies or their mimetics to interact with cell surface markers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If magnetic separation techniques are used to isolate target cells, then cell surface marker-based separation is achieved, but processing time increases significantly (upwards of two hours) and cell viability deteriorates due to shear forces
Solution Approach 1:
The patent employs temperature-responsive hydrogels that undergo phase transition between gel and sol states. At physiological temperature (37°C), the hydrogel is in gel state and binds to target cells via biological moieties. Upon cooling to room temperature, the hydrogel transitions to sol state, releasing the bound cells. This phase transition mechanism enables rapid cell separation and release without requiring prolonged processing or harsh conditions that would compromise cell viability.
Solution Approach 2:
The patent replaces the mechanical magnetic separation system with a thermally-driven hydrogel system. Instead of using magnetic fields and magnetic beads that exert shear forces on cells, the invention uses temperature-controlled hydrogel phase transitions to bind and release cells. This substitution eliminates the mechanical stress inherent in magnetic separation while maintaining marker-specific binding capability through biological moieties attached to the hydrogel.
2Measurement precision
If magnetic particles are used for cell separation, then target cells can be isolated by cell surface markers, but extensive culturing (four to five days) is required to dilute magnetic particles to meet FDA standards, and cell viability is compromised
Solution Approach 1:
The patent uses disposable, biodegradable hydrogel particles instead of persistent magnetic beads. These hydrogels are designed to be used once and then discarded or degraded, eliminating the need for prolonged culturing to dilute magnetic particles. The hydrogels can be easily removed from the cell preparation through temperature-induced phase transition or enzymatic degradation, providing a clean cell product suitable for clinical applications without extended culture requirements.
Solution Approach 2:
The patent changes the physical state parameter of the hydrogel from gel to sol through temperature reduction. This parameter change allows the hydrogel to release bound cells and be easily separated from the cell preparation. Unlike magnetic particles that require dilution through extended culturing, the hydrogel's sol-gel transition provides rapid and complete release, maintaining cell viability and eliminating the four-to-five-day culturing requirement to meet FDA standards.
3Measurement precision
If magnetic separation is performed to achieve specific cell isolation, then target cells are separated by markers, but the process requires manual manipulation and dilution steps that compromise cell health
Solution Approach 1:
The patent implements a self-service system where the temperature-responsive hydrogel automatically binds and releases cells based on temperature changes, without requiring manual intervention for particle removal or cell dilution. The hydrogel performs the separation function autonomously through its inherent phase transition properties, eliminating the need for operator-performed dilution steps and reducing manual manipulation that could compromise cell health.
Solution Approach 2:
The patent introduces temperature as an intermediary control parameter that mediates the binding and release of cells to and from the hydrogel. By controlling temperature, the system automatically regulates hydrogel phase state, which in turn controls cell attachment and release. This intermediary mechanism replaces manual manipulation steps with a simple, non-invasive temperature control process that maintains cell integrity.
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 fast, specific, and viable separation of target cells, maintaining cell integrity and reducing processing time, eliminating the need for magnetic particles, and allowing direct use in immunotherapies, while being suitable for all applications requiring cell surface marker-based separation.
Implementation Method 1
hydrogel technology that can reversibly exist in polymerized and depolymerized states depending on outside conditions
Implementation Method 2
separating said cell-hydrogel complexes from unbound cells and other components of the biological sample in a centrifugation vessel via their sedimentation velocity in a counter-current centrifuge
Implementation Method 3
specific binding of target cells to phase-change hydrogel compositions... binding of the target cells to the hydrogel via said biological moieties
Data Source
Figure 1
AI summary
The present invention relates to methods for the specific separation of target cells from a biological sample, comprising specific binding of the target cells to phase- change hydrogel compositions and separation of respective cell-hydrogel complexes by counter-current centrifugation.