NK Cell Purification via Magnetic Sedimentation
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Solution Overview
Problem
Current cell separation methods are time-consuming, laborious, and inefficient, particularly in separating desired cells from whole blood samples, as they often require initial erythrocyte removal and involve centrifugation steps, which can damage cells and result in low recovery and purity.
Innovation Solution
A magnetically enforced sedimentation method using erythrocyte aggregation reagents like hydroxypropylmethylcellulose and magnetic particles with antigen recognizing moieties to separate cells directly from whole blood samples without initial erythrocyte removal or centrifugation, allowing for rapid and minimal stress cell separation with high recovery and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional cell separation methods are used, then cells can be separated from whole blood, but the process is time-consuming and laborious
Solution Approach 1:
The patent combines erythrocyte aggregation (using reagents like dextran or hydroxyethyl starch) with magnetic particle-based leukocyte depletion into a single simultaneous process. The magnetic particles bind to leukocytes while erythrocytes aggregate and sediment, allowing both functions to occur concurrently rather than sequentially, thereby reducing total separation time and increasing productivity
Solution Approach 2:
The patent replaces the mechanical centrifugation step with a magnetic field-based separation system. Instead of using centrifugal force to separate cells, magnetic particles conjugated to antibodies against leukocyte surface markers are used to selectively bind and remove leukocytes through magnetic forces, eliminating the need for time-consuming centrifugation while maintaining separation effectiveness
2Productivity
If centrifugation steps are used for cell separation, then cells can be separated from plasma, but cell damage occurs and recovery is reduced
Solution Approach 1:
The patent substitutes mechanical centrifugation with magnetic field-based separation. Magnetic particles conjugated to specific antibodies bind to target leukocytes through antigen-antibody interactions, and the magnetic field selectively removes these bound cells without applying mechanical stress to other cell types, thereby preventing cell damage and improving recovery
Solution Approach 2:
The patent introduces magnetic particles as intermediary carriers that mediate the separation process. These particles are conjugated to antibodies that specifically recognize leukocyte surface markers, allowing selective binding and removal of leukocytes without direct mechanical intervention, thus protecting cells from damage while maintaining high recovery rates
3Manufacturing precision
If erythrocyte removal is performed before cell separation, then leukocyte separation is improved, but additional steps are required and time is lost
Solution Approach 1:
The patent merges erythrocyte removal and leukocyte separation into a single simultaneous operation. Erythrocyte aggregation reagents (such as dextran or hydroxyethyl starch) are added to the whole blood sample along with magnetic particles conjugated to anti-leukocyte antibodies. Both processes occur concurrently: erythrocytes aggregate and sediment while magnetic particles bind to and remove leukocytes, eliminating the need for sequential steps and reducing overall complexity
Solution Approach 2:
The patent performs preliminary erythrocyte aggregation and sedimentation before the magnetic separation step, but in a way that prepares the sample for simultaneous processing rather than requiring separate operations. The aggregation reagent pre-treats the sample to facilitate subsequent magnetic particle binding and leukocyte removal, streamlining the overall process while maintaining high purity
4Manufacturing precision
If multiple separation steps are used, then cell purity is improved, but the process becomes more laborious and time-consuming
Solution Approach 1:
The patent combines multiple separation functions into a single operational step: erythrocyte aggregation, leukocyte binding via magnetic particles, and simultaneous sedimentation all occur in one mixture. This integrated approach achieves high cell purity without requiring multiple sequential operations, thereby reducing labor intensity and simplifying the overall process while maintaining or improving separation effectiveness
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 accelerates cell separation, reduces stress on cells, and improves recovery and purity by simultaneously aggregating erythrocytes and immobilizing other cellular components using magnetic forces, enabling efficient cell preparation for further processing without the need for centrifugation.
Implementation Method 1
The sedimentation of cells in a sample is accelerated if simultaneously (1) the erythrocytes are aggregated by an reagent (e.g. rouleaux-forming agent), i.e. an erythrocyte aggregation reagent such as hydroxypropylmethylcellulose (HPMC) and (2) one or more cellular components
Implementation Method 2
non-ionic polymers such as polysaccharides and synthetic polymers promote red blood cells, i.e. erythrocytes, aggregation when infused in vivo or added to suspensions of erythrocytes in buffer or plasma in vitro
Implementation Method 3
one or more cellular components, i.e. other blood cells and/or thrombocytes, are bound specifically by magnetic particles, wherein a magnetic field gradient is applied to the magnetic particles, immobilizing the cells bound to the magnetic particles
Data Source
AI summary
The present invention provides methods and compositions for separating cells from a sample containing erythrocytes. The method is for recovering desired cells from a sample containing the desired cells, erythrocytes and undesired cells comprising: a) contacting the sample with a composition, said composition comprising: i) an erythrocytes aggregation reagent ii) at least one antigen recognizing moiety coupled to a magnetic particle, wherein said particle with said at least one antigen recognizing moiety specifically binds to at least one antigen specific for one or more undesired cellular components; b) applying simultaneously i) gravity sedimentation for sedimentation of erythrocytes and ii) a magnetic field gradient to said sample for immobilizing said magnetic particle generating a pellet and a supernatant phase, and c) recovering the desired cells from the supernatant phase. Compositions for the use within the present method are also disclosed.