Microvesicle Extraction by Electrostatic Aggregation and Filtration
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Solution Overview
Problem
Conventional methods for isolating microvesicles, such as ultracentrifugation, size exclusion, immunoaffinity isolation, and polymeric methods, face challenges including low yield, high cost, long processing times, and low purity, making them unsuitable for practical applications, especially in molecular diagnostics.
Innovation Solution
A method involving the use of a polyvalent cationic substance to aggregate microvesicles via electrical force, followed by capture through a filter and separation using an elution solution, eliminating the need for centrifugation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ultracentrifugation isolation method is used, then reliability of microvesicle isolation is improved, but productivity is worsened due to long processing time and low yield
Solution Approach 1:
The patent replaces the mechanical ultracentrifugation system with a chemical aggregation system using polyvalent cationic substances. Instead of using high-speed rotation to separate microvesicles, the invention uses electrostatic interactions between cationic polymers and anionic microvesicle surfaces to form aggregates that can be easily filtered, thereby eliminating the need for expensive ultracentrifuges and reducing processing time while maintaining isolation reliability
Solution Approach 2:
The patent changes the physical-chemical parameters of the isolation process by adjusting the charge interactions between microvesicles and polymers. By controlling the charge density, polymer concentration, and ionic strength, the method transforms microvesicles from a state requiring mechanical separation to a state amenable to simple filtration, significantly improving productivity while maintaining reliability
2Reliability
If ultracentrifugation isolation method is used, then isolation reliability is improved, but loss of time is worsened due to processing time of 8 hours or more
Solution Approach 1:
The patent applies preliminary chemical action by pre-aggregating microvesicles with polyvalent cationic substances before filtration. This preliminary aggregation step transforms the microvesicles into larger, filterable structures, eliminating the need for time-consuming ultracentrifugation and reducing the overall processing time from 8 hours to a much shorter duration while maintaining isolation reliability
3Manufacturing precision
If size exclusion method is used, then manufacturing precision of microvesicle isolation is improved, but loss of substance is worsened due to microvesicles sticking to filter
Solution Approach 1:
The patent introduces polyvalent cationic substances as intermediaries that bind to microvesicles and prevent their adhesion to the filter surface. These cationic polymers act as protective mediators, allowing microvesicles to pass through the filter without sticking, thereby recovering nearly all microvesicles while still achieving effective separation of impurities, thus improving both yield and purity
4Manufacturing precision
If immunoaffinity isolation method is used, then manufacturing precision of specific microvesicle isolation is improved, but loss of time is worsened due to long antibody production process
Solution Approach 1:
The patent employs polyvalent cationic substances that serve as universal reagents capable of binding to all anionic microvesicles regardless of their specific origin or surface markers. This universal approach eliminates the need for time-consuming antibody production while still achieving effective isolation, making the method broadly applicable to various microvesicle types without sacrificing selectivity
5Productivity
If polymeric method using PEG is used, then productivity of microvesicle sedimentation is improved, but manufacturing precision is worsened due to co-precipitation of impurities
Solution Approach 1:
The patent applies local quality differentiation by using polyvalent cationic substances that selectively interact with anionic microvesicles through electrostatic forces, while leaving other impurities unaffected. This selective interaction ensures that only microvesicles are aggregated and precipitated, while proteins and other contaminants remain in solution, thereby achieving both high productivity and high purity without co-precipitation
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 enhances microvesicle yield and purity while reducing processing time and cost, facilitating their use in molecular diagnostics.
Implementation Method 1
adding a polyvalent cationic substance to the biological sample to form an aggregate in which the microvesicles and the polyvalent cationic substance are aggregated with other via an electrical force
Implementation Method 2
passing the biological sample containing the aggregate through a capture filter while the aggregate is captured by the capture filter
Implementation Method 3
passing an elution solution through the capture filter where the aggregate is captured such that the microvesicles are separated from the aggregate
Data Source
AI summary
The present invention relates to a method for extracting microvesicles from a biological sample, the method comprising the steps of: adding a polyvalent cationic material to the biological sample to form an aggregate in which the microvesicles and the polyvalent cationic material are aggregated by electrical force; capturing the aggregate by a capture filter while the biological sample including the aggregate passes through the capture filter; and extracting the microvesicles by allowing an elution solution to pass through the capture filter with the aggregate captured therein to isolate the microvesicles from the aggregate. Accordingly, microvesicles may be extracted using a polyvalent cationic material, without a centrifugation process.