Centrifugal Microfluidic Isolation of EVs and Cell-Free Nucleic Acids
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Solution Overview
Problem
Existing methods for isolating different analyte classes from a biological sample require large sample volumes and multiple steps, leading to increased costs and patient burden, and result in the destruction of extracellular vesicles, preventing further analysis of their characteristics.
Innovation Solution
A centrifugal microfluidic system with a fluidic module containing separate isolation chambers and structures, using size-based filtration and magnetizable particles, allows simultaneous isolation of extracellular vesicles and cell-free nucleic acids from a single sample volume, enabling an integrated process chain.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If multiple analyte classes are isolated from the same sample volume using separate devices and multiple steps, then the isolation completeness of each analyte class is improved, but the device complexity and operational complexity increase significantly
Solution Approach 1:
The patent combines multiple isolation functions (EV isolation, CTC isolation, cfDNA isolation) into a single integrated centrifugal microfluidic device. The device contains multiple isolation chambers with different isolation structures that can simultaneously process different analyte classes from the same sample volume, eliminating the need for multiple separate devices and manual transfer steps.
Solution Approach 2:
The centrifugal microfluidic device is designed as a universal platform that can isolate multiple types of analytes (extracellular vesicles, circulating tumor cells, and cell-free DNA) using a single device. Each isolation chamber is equipped with specific isolation structures tailored for different analyte classes, allowing the device to perform multiple functions simultaneously.
2Reliability
If multiple isolation steps are performed sequentially using separate devices, then the purity of each analyte class is improved, but the processing time and loss of substance increase
Solution Approach 1:
The patent enables continuous processing of multiple analyte classes in a single integrated workflow. The centrifugal force continuously drives the sample through different isolation chambers, allowing simultaneous isolation of EVs, CTCs, and cfDNA without interrupting the processing flow. This eliminates the time losses associated with manual sample transfer and device switching.
Solution Approach 2:
Multiple isolation procedures that were previously performed sequentially in separate devices are merged into a single parallel processing system. The device uses centrifugal separation to simultaneously isolate different analyte classes in different chambers, reducing total processing time while maintaining purity through dedicated isolation structures for each analyte type.
3Productivity
If phenol-based lysis reagents are used to extract nucleic acids from microvesicles, then the nucleic acid extraction efficiency is improved, but the extracellular vesicles are destroyed and cannot be further analyzed
Solution Approach 1:
The patent divides the sample processing into separate segments or chambers: one chamber isolates intact extracellular vesicles for further analysis, while another chamber processes the same sample to extract nucleic acids. This segmentation allows both goals to be achieved simultaneously without compromising vesicle integrity in the first chamber, as the harsh lysis reagents are confined to the nucleic acid extraction chamber only.
Solution Approach 2:
The centrifugal microfluidic system acts as an intermediary that separates the sample into different processing pathways. The sample is divided into fractions that are directed to different isolation chambers, where appropriate processing conditions are applied. This intermediary system enables selective extraction of nucleic acids from a subset of the sample while preserving the integrity of extracellular vesicles in another fraction.
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 isolation of multiple analyte classes from a small sample volume without destruction, providing separate samples for detailed analysis and reducing the need for multiple devices and manual steps.
Implementation Method 1
The first isolation structure is a filter with a pore size in a range of 20 nanometers to 200 nanometers, preferably in a range of 20 nanometers to 45 nanometers
Implementation Method 2
The second isolation structure is a surface for binding analytes of the second analyte class, and wherein the surface is formed by magnetizable particles
Implementation Method 3
A centrifugal microfluidic system is defined as a system designed to handle liquids by utilizing the centrifugal force generated during rotation
Data Source
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AI summary
In a method for isolating analytes of a first analyte class, which are extracellular vesicles and/or circulating tumour cells, and analytes of a second analyte class, which are cell-free nucleic acids, from the same sample volume of a biological sample in a centrifugal microfluidic system, the sample volume is guided into a first isolation chamber of a fluidic module, this first isolation chamber containing a first isolation structure, so that the analytes of the first analyte class are retained by the first isolation structure, while the analytes of the second analyte class are not retained by the first isolation structure and pass through the first isolation structure as part of a residual liquid. The residual liquid is guided into a second isolation chamber of the fluidic module, this second isolation chamber containing a second isolation structure, so that the analytes of the second analyte class are retained by the second isolation structure. The analytes of the first analyte class and of the second analyte class are separated from the respective isolation structure in order to provide the analytes for a subsequent analysis.