Radial Particle Separation Apparatus for DNA Mutation Isolation
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Solution Overview
Problem
Current methods for purifying and separating molecules, such as DNA, face challenges in efficiently isolating genetic and epigenetic differences in heterogeneous samples, particularly in identifying specific mutations or epigenetic variations amidst a preponderance of similar particles.
Innovation Solution
A particle separation apparatus with a central reservoir surrounded by electrodes and a separation medium, utilizing a time-varying driving field and a mobility-altering field to selectively move target particles based on their affinity to immobilized agents, allowing for the extraction of DNA with point mutations or epigenetic variations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional purification methods are used, then the purification process can be performed with simple equipment, but the ability to efficiently isolate genetic and epigenetic differences in heterogeneous samples is insufficient
Solution Approach 1:
The separation apparatus is divided into multiple independent arms (typically 3-6 arms) arranged radially around a central collection reservoir. Each arm contains a stationary phase with immobilized affinity ligands and is independently controlled by electrodes. This segmentation allows simultaneous separation of different particle types while maintaining a relatively simple overall structure.
Solution Approach 2:
The system employs dynamic control of electric field strength and direction through independently controlled electrodes in each arm. By varying the voltage applied to each arm in a coordinated sequence, particles are dynamically directed toward the central reservoir based on their affinity characteristics. This dynamic control enables high specificity separation without requiring complex mechanical moving parts.
2Measurement precision
If multiple particle types are present in the sample, then the sample represents a realistic heterogeneous mixture, but the isolation of specific target particles becomes more difficult
Solution Approach 1:
Each arm of the separation apparatus is equipped with stationary phase containing specific affinity ligands that selectively bind to particular particle types. This local functional differentiation allows the system to simultaneously recognize and separate multiple different particle types present in the heterogeneous sample, with each arm optimized for specific target particles.
Solution Approach 2:
The system exploits changes in particle mobility that occur when particles bind to affinity ligands in the stationary phase. By applying electric fields and monitoring mobility changes, the apparatus can distinguish between different particle types based on their specific binding characteristics, enabling precise isolation even from complex heterogeneous mixtures.
3Speed
If SCODA-based transport is used, then net motion of molecules can be achieved through synchronized driving force, but the separation of particles with similar characteristics remains challenging
Solution Approach 1:
Different arms are equipped with stationary phases having different affinity characteristics tailored to specific particle types. This local differentiation allows particles with similar characteristics to be separated based on their differential binding affinities, while still maintaining efficient net motion through the SCODA mechanism in each arm.
Solution Approach 2:
The system employs periodic alternation of electric field directions and strengths across multiple arms in a coordinated sequence. This periodic action enhances the SCODA effect by creating synchronized back-and-forth motion that results in net directional transport, while the varying field parameters across different arms provide resolution between similar particle types.
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
Enables the effective isolation of target DNA sequences with mutations or epigenetic differences from wild-type DNA, facilitating genotyping and identifying fetal genetic material in maternal samples with high specificity and efficiency.
Implementation Method 1
The separation medium may include an affinity agent that has a binding affinity for a targeted particle
Implementation Method 2
SCODA is an approach that may be applied for purifying, separating, or concentrating particles. SCODA may be applied, for example, to DNA, RNA and other molecules including proteins and polypeptides.
Implementation Method 3
SCODA is an approach that may be applied for purifying, separating, or concentrating particles. SCODA may be applied, for example, to DNA, RNA and other molecules including proteins and polypeptides.
Implementation Method 4
The apparatus includes at least three electrodes circumferentially surrounding a central reservoir with a separation medium between at least one electrode and the central reservoir
Data Source
AI summary
The invention provides apparatus for separation of particles and methods for using the apparatus. In an embodiment, the apparatus includes three arms extending radially from a central reservoir, each arm being associated with a separation electrode. At least one on the arms includes a separation medium. Using a sequence of driving and mobility-changing voltages, target particles can be separated from closely related particles within a sample. For example, single point mutations can be resolved from a sample containing predominantly wild type nucleic acids.