Cell Separation by Nucleic Acid Labeling in Heterogeneous Samples
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing cell isolation methods struggle with sensitivity when dealing with heterogeneous samples containing similar cell populations, such as fetal and maternal erythroblasts, often leading to loss of cell integrity and inefficiency in separation.
Innovation Solution
An isothermal amplification method is used to label target nucleic acids within specific cell populations, allowing for the separation of these cells based on the presence of a unique nucleic acid marker, using fluorescent or magnetic labels to facilitate detection and isolation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional cell isolation methods (Immunomagnetic Cell Separation, FACS, Density Gradient Centrifugation) are used on heterogeneous samples with similar cell populations, then cell separation is achieved, but sensitivity is lost and cell integrity is compromised
Solution Approach 1:
The method segments the cell population identification process by targeting specific nucleic acid sequences unique to each cell type. Instead of relying on surface markers or physical properties that may be similar across cell populations, the invention divides the detection space at the molecular level, using cell-type-specific gene expression profiles to distinguish between fetal and maternal cells, thereby maintaining sensitivity without compromising cell integrity.
Solution Approach 2:
The invention introduces nucleic acid amplification and labeling as an intermediary step between cell sampling and separation. By converting the invisible molecular differences (gene expression) into visible labels (fluorescent or magnetic markers), the method enables precise identification and separation of similar cell populations while keeping cells intact and viable throughout the process.
2Manufacturing precision
If conventional cell separation methods are applied to differentiate similar cell populations (e.g., fetal and maternal erythroblasts), then separation is attempted, but the ability to differentiate is lost due to similar physical and chemical properties
Solution Approach 1:
The invention replaces mechanical and physical separation methods (based on size, density, or surface markers) with a molecular recognition system. By using nucleic acid hybridization and amplification to detect cell-type-specific genes, the method substitutes physical differentiation with molecular differentiation, enabling precise separation of cells that have identical physical properties but different gene expression profiles.
Solution Approach 2:
The method changes the detection parameter from physical properties (size, density, surface antigens) to molecular properties (gene expression patterns). By amplifying and labeling specific nucleic acid sequences that are unique to fetal or maternal cells, the invention transforms an undetectable difference into a detectable signal, enabling efficient separation based on molecular rather than physical parameters.
3Productivity
If reagents and conditions are used to separate similar cell populations, then separation may be achieved, but cell integrity is lost
Solution Approach 1:
The invention enables cells to 'self-label' through intracellular nucleic acid amplification. The amplification reaction occurs inside the cell using the cell's own machinery and reagents, creating labels that are inherently associated with the cell without requiring external attachment steps that could damage the cell membrane or structure. This self-service approach maintains cell integrity while achieving efficient separation.
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 effectively separates similar cell populations while preserving cell integrity, enabling efficient isolation of fetal cells from maternal samples for non-invasive prenatal diagnosis.
Implementation Method 1
using fluorescent or magnetic labels to facilitate detection and isolation
Implementation Method 2
using fluorescent or magnetic labels to facilitate detection and isolation
Implementation Method 3
An isothermal amplification method is used to label target nucleic acids within specific cell populations
Data Source
Figure 1
Figure 2A
Figure 2B
AI summary
The present invention provides an in vitro method for separating a first population of cells from a second population of cells comprised in a sample, wherein the first population of cells is characterized in that they comprise at least one target nucleic acid molecule that is not present in the second population of cells.