Single Fetal Cell Isolation via Negative Selection and Mouth Pipetting
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Solution Overview
Problem
Current methods for isolating fetal cells from maternal blood face challenges in obtaining cells with complete genomes due to low specificity of antibodies and vulnerability of fetal cells, leading to incomplete genome sequences and difficulty in detecting gene mutations.
Innovation Solution
A method involving immunomagnetic bead sorting using antibodies specific to non-target cells, followed by flow cytometry and mouth-controlled pipette isolation, to obtain a single fetal cell with a complete genome, suitable for whole genome amplification and sequencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If immunomagnetic bead sorting uses antibodies specific to non-target cells, then the purity of isolated fetal cells is improved, but the complexity of the isolation process increases
Solution Approach 1:
Instead of using antibodies that directly bind to fetal cells (positive selection), the patent uses antibodies that bind to maternal cells (negative selection), thereby isolating fetal cells by removing unwanted cells. This inversion approach improves purity while managing complexity through clever antibody selection strategy
Solution Approach 2:
The patent introduces magnetic beads as intermediaries that carry antibodies against maternal cells. These beads serve as mediators between the antibody and the maternal cells, enabling easy separation through magnetic fields while maintaining the specificity of antibody binding for pure fetal cell isolation
2Manufacturing precision
If multiple sorting steps are performed to obtain complete genome, then the completeness of genome sequence is improved, but the vulnerability of fetal cells to damage increases
Solution Approach 1:
The patent extracts and removes maternal cells from the blood sample through immunomagnetic bead sorting, isolating fetal cells in a single primary sorting step. This extraction approach achieves complete genome sequences by eliminating contamination from maternal cells without requiring multiple manipulative sorting steps that could damage fragile fetal cells
Solution Approach 2:
The patent performs preliminary enrichment of fetal cells by removing maternal cells before subsequent analysis. This preliminary action ensures that fetal cells are isolated in a clean state with complete genomes, avoiding the need for repeated sorting operations that could compromise 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 method effectively isolates a single fetal cell with a complete genome, enabling accurate whole genome sequencing and analysis for determining fetal conditions such as gender, genetic relationships, and chromosomal abnormalities.
Implementation Method 1
subjecting the blood sample to immunomagnetic bead sorting, thereby obtaining a first sorted product containing the target cell, wherein the immunomagnetic bead sorting uses beads coated with an antibody
Implementation Method 2
incubating the first solution containing the target cell with Hoechst 33342/Propidium Iodide (PI) dyes at room temperature in a dark place, thereby obtaining a second solution containing stained cells
Data Source
AI summary
The present invention provides a method for obtaining a target cell from a blood sample and a use thereof. The method comprises: (1) subjecting the blood sample to immunomagnetic bead sorting to obtain a first sorted product comprising the target cell; (2) subjecting the first sorted product to flow cytometric sorting to obtain a second sorted product comprising the target cell; and (3) separating the second sorted product by a mouth pipetting method to obtain the target cell in a unicellular form.