miRNA Extraction from Extracellular Vesicles Using Capture Device
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Solution Overview
Problem
Current methods for extracting miRNA from extracellular vesicles (EVs) in sample solutions, such as ultracentrifugation and aggregation reagent methods, are inefficient and result in sample loss, requiring multiple steps and increasing the risk of EV discarding during the separation process.
Innovation Solution
A method involving a device capable of capturing EVs, where the sample solution is contacted with the device to capture EVs, followed by direct disruption with a solution to extract miRNA, eliminating the need for EV separation steps, using a device made of materials resistant to disruption solutions like cellulose nanofibers or nanowires.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ultracentrifugation is used to collect EVs from sample solution, then EVs can be separated and collected, but the work procedure increases and sample loss occurs during fraction collection
Solution Approach 1:
The patent extracts only the necessary EV-containing fraction after ultracentrifugation, discarding unnecessary fractions. This minimizes sample loss by focusing collection efforts only on the relevant EV-containing portion rather than processing entire sample volumes through multiple collection steps.
Solution Approach 2:
The patent performs preliminary ultracentrifugation to concentrate EVs into specific fractions before final collection. By pre-concentrating EVs in the pellet or specific density fractions, the method ensures high recovery efficiency while minimizing losses during subsequent handling.
2Reliability
If aggregation reagent method is used to separate EVs, then EVs can be aggregated and separated, but the work procedure increases and sample loss occurs during separation operation
Solution Approach 1:
The patent uses aggregation reagents as intermediaries to facilitate EV separation. These reagents temporarily bind to EVs, enabling their separation from sample matrix, and are subsequently removed. This intermediary approach maintains EV integrity while enabling efficient separation with minimal loss.
Solution Approach 2:
The patent discards the aggregation reagent after it has served its separation function, while recovering the EVs in purified form. This single-step separation approach recovers EVs without the multiple handling steps that cause loss in conventional methods.
3Reliability
If multiple separation steps are used to collect EVs, then EV purification can be achieved, but the work procedure increases and time consumption increases
Solution Approach 1:
The patent combines EV concentration and purification into a single ultracentrifugation step, eliminating the need for sequential separation operations. By concentrating EVs directly into a pellet or specific density fraction, the method achieves both concentration and purification simultaneously, dramatically reducing processing time.
Solution Approach 2:
The ultracentrifugation step serves multiple functions simultaneously: it concentrates EVs, purifies them from sample matrix, and prepares them for downstream analysis. This multi-functional approach replaces multiple specialized separation steps, reducing overall procedure time while maintaining purification quality.
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 simplifies the extraction process, reduces sample loss, and allows for the analysis of trace amounts of miRNA, particularly from small sample volumes like saliva, enhancing the efficiency and accuracy of miRNA analysis.
Implementation Method 1
a device capable of capturing extracellular vesicles... an extracellular vesicle capture step of bringing the sample solution into contact with the device, to capture an extracellular vesicle in the device
Implementation Method 2
a miRNA extraction step of bringing the device that captured the extracellular vesicle with the disruption solution of extracellular vesicles, to disrupt the extracellular vesicle and extract miRNA from the extracellular vesicle into the disruption solution
Data Source
AI summary
The present invention provides a novel miRNA extraction method and a method for analyzing miRNA extracted by using said miRNA extraction method. According to the present invention, provided is, for example, a method for extracting miRNA from extracellular vesicles in a sample solution, by using a device capable of capturing extracellular vesicles, the miRNA extraction method comprising: an extracellular vesicle capturing step for capturing extracellular vesicles in a sample solution onto a device by bringing the sample solution and the device in contact with each other; and a miRNA extraction step for homogenizing the extracellular vesicles by bringing the device having captured the extracellular vesicles in contact with a homogenization liquid for extracellular vesicles to extract miRNA from the extracellular vesicle into the homogenization liquid.