Target Cell Separation Using Filter-Retained Ligand Microparticles
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Solution Overview
Problem
Current cell separation technologies fail to efficiently isolate target cells based on their surface antigens without using magnetic particles or antibody-coated beads, leading to impurities and reduced cell viability, which is crucial for cell therapies.
Innovation Solution
A method using non-porous microparticles with covalently bound capture ligands that specifically bind to cell surface molecules, followed by incubation, washing, and mechanical dissociation to achieve high purity and viability of target cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If magnetic particle separation is used to isolate target cells, then cell separation efficiency is improved, but residual magnetic nanoparticles remain in the final cell suspension and require bulky, expensive equipment
Solution Approach 1:
The patent extracts the harmful magnetic nanoparticles from the separation system by using non-magnetic porous particles instead. The capture ligands are immobilized on the particle surface, and after separation, the particles are removed by filtration, ensuring no magnetic residues remain in the final cell suspension.
Solution Approach 2:
The patent employs disposable non-magnetic porous particles that are used once and then discarded through filtration. This eliminates the need for expensive magnetic separation equipment and ensures complete removal of particles from the final product, as each new cartridge contains fresh particles without carryover contamination.
2Productivity
If chemical elution buffers are used to release target cells from magnetic particles, then cell elution is achieved, but cell viability is reduced
Solution Approach 1:
The patent replaces chemical elution with mechanical separation. Non-magnetic porous particles are used that can be physically filtered out, allowing cells to be released without exposure to harsh chemical buffers. The particles are removed by filtration through a filter with appropriate pore size, eliminating the need for chemical elution and preserving cell viability.
3Manufacturing precision
If non-porous microparticles with covalently immobilized capture ligands are used, then separation purity is improved, but the complexity of particle functionalization increases
Solution Approach 1:
The patent uses porous particles with controlled pore sizes that prevent particle passage through filtration while allowing cell-microparticle complexes to be retained and then dissociated. The porosity enables efficient capture ligand immobilization on the internal surface area, achieving high separation purity without requiring overly complex functionalization procedures.
4Manufacturing precision
If extensive washing steps are performed to remove residual buffers and antibodies, then product purity is improved, but processing time increases
Solution Approach 1:
The patent extracts the need for extensive washing steps by using non-magnetic porous particles that are completely removed by filtration. Since the particles themselves are discarded rather than eluted, residual buffers and antibodies are minimized at the source, reducing the need for multiple washing steps and decreasing overall processing time.
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 method ensures high purity and viability of target cells by eliminating the need for chemical elution buffers and magnetic particles, reducing processing time and maintaining cell health.
Implementation Method 1
contacting a sample comprising a suspension of viable target cells that display a molecule on the cell surface with non-porous microparticles that have a density of about 1.45 g/cm3 or greater, a diameter of about 10 μm to 200 μm, and a capture ligand covalently immobilized to the microparticle surface that is capable of specifically binding to said molecule
Implementation Method 2
separating non-bound substances in said sample from said target cell/microparticle complex by washing said non-bound substances through a filter while retaining said target cell/microparticle complex
Implementation Method 3
mechanically dissociating said target cell/microparticle complex and eluting said viable target cells through said filter while retaining said microparticles with said capture ligand covalently immobilized to the microparticle surface
Data Source
AI summary
The invention relates to a method for separating viable target cells from a sample comprising the steps of contacting a sample comprising a suspension of viable target cells that display a molecule on the cell surface with non-porous microparticles that have a density of about 1.45 g/cm3 or greater, a diameter of about 10 μm to 200 μm, and a capture ligand covalently immobilized to the microparticle surface that is capable of specifically binding to said molecule; incubating said sample without substantial agitation to form a target cell/microparticle complex; separating non-bound substances in said sample from said target cell/microparticle complex by washing said non-bound substances through a filter while retaining said target cell/microparticle complex; mechanically dissociating said target cell/microparticle complex and eluting said viable target cells through said filter while retaining said microparticles with said capture ligand covalently immobilized to the microparticle surface as well as a cartridge, kit-of-parts, an apparatus configured to be used in the method and a medicament comprising viable target cells obtainable by the method.