Cell Purification Column Using Affinity Capture
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Solution Overview
Problem
Current methods for purifying biological cells, such as those using magnetic beads, often result in cell damage, low yields, and impure populations due to physical stress and lengthy processing times, failing to efficiently capture and recover viable cells.
Innovation Solution
A column-based method where cells are captured and purified using a solid phase with a bed of medium, allowing for rapid and reversible capture and elution of viable cells, utilizing unidirectional or bidirectional flow, and employing capture entities like antibodies or aptamers to target specific cells, while maintaining cell viability and purity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If magnetic beads are used to capture and purify cells, then cell capture capability is improved, but cell viability deteriorates due to physical stress from magnetic capture and cell clumping
Solution Approach 1:
The patent replaces the mechanical magnetic capture system with a chromatography-based capture system using antibodies or affinity ligands immobilized on column media. This substitution eliminates the need for magnetic fields and mechanical manipulation of bead-cell complexes, thereby preventing cell damage while maintaining capture capability through specific biological interactions.
Solution Approach 2:
The patent introduces an intermediary capture mechanism using antibodies or affinity ligands as mediators between the column media and target cells. This intermediary approach allows specific cell capture through molecular recognition rather than direct mechanical or magnetic forces, preserving cell integrity while achieving high capture efficiency.
2Reliability
If magnetic bead methods are used for cell purification, then cell capture is achieved, but processing time increases and cell yield decreases due to lengthy processing and repeated manipulation
Solution Approach 1:
The patent implements continuous flow chromatography where cells pass through the column in a continuous stream, allowing capture to occur during the flow process rather than requiring batch processing with repeated loading and washing steps. This continuous action significantly reduces processing time while maintaining capture efficiency.
Solution Approach 2:
The patent enables rapid cell capture by optimizing flow rates and column parameters to allow cells to pass through quickly while still achieving effective capture. The method skips the lengthy incubation and repeated washing steps required by magnetic bead methods, rushing through the capture process in a single pass or minimal passes.
3Reliability
If magnetic beads are used for cell purification, then cell separation is achieved, but cell purity deteriorates due to cell clumping and contamination
Solution Approach 1:
The patent replaces mechanical mixing and magnetic separation that cause cell clumping with a gentle flow-through chromatography system. Cells flow individually through the column and are captured one-by-one or in small groups, preventing the formation of large clumps and ensuring homogeneous cell populations with high purity.
4Manufacturing precision
If repeated magnetic capture is performed to purify cells, then purification efficiency is improved, but cell viability deteriorates due to repeated physical stress
Solution Approach 1:
The patent achieves high purification efficiency in a single continuous flow-through pass through the column, eliminating the need for repeated capture and release cycles. The continuous process maintains cell viability by avoiding repeated mechanical stress while achieving the same or better purification through optimized column parameters and flow rates.
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 method effectively captures and recovers viable cells with high purity and efficiency, reducing cell damage and processing time, enabling rapid cell isolation and utilization as a stationary phase for chromatography, while maintaining cell integrity and viability.
Implementation Method 1
the solid phase can be further comprised of a capture entity capable of capturing cells present in the biological sample
Implementation Method 2
The sample can be passed through the column using unidirectional or bidirectional flow
Data Source
AI summary
This invention relates to devices and methods for purifying, detecting and using biological cells. A variety of cell types including viable tumor, stem, immune and sperm cells can be purified from a complex biological sample using a column, including a pipette tip column. Methods of the invention can aid research, diagnosis and treatment of cancer. Purified viable cells can be detected on the column or eluted from the column and detected. Cells on a column can be used as a stationary phase for liquid chromatography. Cells may be removed, recovered and analyzed.