EV Capture Microparticles with Cleavable Linkers for Virus Isolation
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Solution Overview
Problem
Current methods are inadequate for reliably detecting, isolating, and characterizing extracellular vesicles and viruses in biofluids due to similarities in size and structure, which complicates their differentiation and identification for diagnostic and research purposes.
Innovation Solution
A method involving functionalized capture microparticles and nanoparticles with specific binding agents, using linkers that can be cleaved to separate target extracellular vesicles from non-target ones, followed by techniques like dielectrophoresis or centrifugation for purification, allowing for precise isolation and characterization of target EVs or viruses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional isolation methods are used for extracellular vesicles and viruses, then isolation can be achieved, but reliable detection and characterization are compromised due to similarities in size and structure
Solution Approach 1:
The patent introduces functionalized capture particles as intermediary agents that specifically bind to target EVs or viruses through surface markers. These capture particles serve as mediators between the sample and detection systems, enabling reliable identification and characterization despite the similar physical properties of different vesicle types
Solution Approach 2:
The patent applies local quality by functionalizing specific regions of capture particles with targeted binding agents that recognize particular surface markers on EVs or viruses. This localized functionalization enables specific differentiation and characterization of target entities based on their unique surface properties rather than relying solely on overall size and structure
2Measurement precision
If functionalized capture particles with cleavable linkers are used, then precise isolation of target EVs is achieved, but the process complexity increases
Solution Approach 1:
The patent segments the isolation process into distinct functional modules: capture particles with cleavable linkers for specific binding, separation techniques for physical isolation, and release mechanisms for recovering pure target EVs. This segmentation enables precise isolation while making the complex process more controllable and reproducible
Solution Approach 2:
The patent utilizes parameter changes, specifically the cleavage of linkers under controlled conditions, to transition target EVs from a bound state during isolation to a free state for recovery. This parameter change enables precise isolation while providing a controlled mechanism to simplify the process at each stage
3Manufacturing precision
If separation techniques like dielectrophoresis or centrifugation are applied, then target EVs are separated from non-target EVs, but the integrity of isolated particles may be compromised
Solution Approach 1:
The patent applies preliminary action by pre-functionalizing capture particles with cleavable linkers and specific binding agents before the separation process. This preliminary preparation enables the use of gentle separation techniques that maintain particle integrity while achieving high purity separation through the pre-established specific binding interactions
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 enables efficient separation and characterization of target extracellular vesicles or viruses from biofluids, facilitating their use as biomarkers for disease detection and diagnosis while maintaining the integrity of the isolated particles for further analysis.
Implementation Method 1
cleaving the plurality of first linkers to dissociate at least the plurality of target EVs from the EV capture microparticles
Implementation Method 2
applying at least one of dielectrophoresis or centrifugation to extract the plurality of target capture nanoparticles
Implementation Method 3
applying at least one of dielectrophoresis or centrifugation to extract the plurality of target capture nanoparticles
Implementation Method 4
functionalised, via a plurality of first linkers, with EV-specific binding agents specific to an EV surface marker, such that a plurality of bound microparticle-EV assemblies is formed
Data Source
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AI summary
A method of isolating target extracellular vesicles, EVs or target viruses from a biofluid comprising a plurality of target EVs and target viruses and non-target EVs and non-target viruses, comprising: introducing a plurality of EV or viral capture microparticles to the biofluid to obtain a precursor mixture, wherein the plurality of EV or viral capture microparticles are functionalised, via a plurality of first linkers, with EV or viral-specific binding agents specific to an EV or first viral surface marker, such that a plurality of bound microparticle-EV and bound viral-microparticle assemblies is formed in the precursor mixture; extracting the bound microparticle-EV/microparticle-viral assemblies; introducing a plurality of target or viral capture nanoparticles, wherein the plurality of target or viral capture nanoparticles are functionalised with target or viral-specific binding agents receptive to surface markers comprised on the target EVs or a second viral surface marker, such that the target or viral capture nanoparticles bind to the plurality of target EVs or target viruses; cleaving the plurality of first linkers to dissociate at least the plurality of target EVs or target viruses from the EV or viral capture microparticles; after the cleaving, extracting the plurality of EV or viral capture microparticles and applying at least one of dielectrophoresis or centrifugation to extract the plurality of target or viral capture nanoparticles, such that the target EVs or viruses are separated from the non-target EVs or non-target viruses.