Antibody Immunolipoplex Biochip for Single EV Detection
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Solution Overview
Problem
Current methods for detecting extracellular vesicles (EVs) in body fluids are time-consuming, expensive, and lack sensitivity, as they require multiple steps and cannot differentiate between EVs from normal and disease cells, making early disease detection challenging.
Innovation Solution
The development of antibody immunolipoplex nanoparticle (Ab-ILN) and antibody tethered lipoplex nanoparticle (Ab-TLN) biochips that use molecular probes to directly detect intra-vesicular DNA/RNA/proteins and membrane proteins within individual EVs, allowing for simultaneous capture and identification of EVs, viruses, and pathogens without amplification steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple separate methods are used for EV isolation and nucleic acid/protein detection, then detection comprehensiveness is improved, but detection time and process complexity increase significantly
Solution Approach 1:
The patent combines EV isolation, nucleic acid detection, and protein detection into a single integrated biochip platform. The biochip surface is functionalized with both antibody capture probes and nucleic acid probes simultaneously, allowing parallel detection of multiple EV components without sequential processing steps.
Solution Approach 2:
The biochip is designed with multi-functional capabilities to perform both EV capture and intra-EV content detection on the same platform. The universal design allows simultaneous analysis of diverse EV components including proteins, mRNA, miRNA, and other nucleic acids through integrated probe systems.
2Quantity of substance
If total nucleic acids and proteins from all EVs are analyzed, then sample quantity is improved, but detection sensitivity for disease-specific EVs decreases
Solution Approach 1:
The biochip employs localized probe functionalization where specific regions of the chip surface are equipped with disease-specific capture antibodies and corresponding detection probes. This localized approach enables targeted enrichment and detection of rare disease-specific EVs while maintaining the ability to analyze total EV content in parallel.
3Quantity of substance
If conventional EV isolation methods are used, then EV separation is achieved, but intra-EV content detection capability is lost
Solution Approach 1:
The biochip performs preliminary capture of target EVs on the chip surface before lysis and nucleic acid/protein detection. This preliminary localization of EVs on the functionalized surface preserves intra-EV content integrity and enables subsequent in-situ detection without requiring external isolation steps that would separate EVs from their contents.
4Measurement precision
If amplification steps are included in detection, then detection sensitivity is improved, but detection time and process complexity increase
Solution Approach 1:
The patent employs highly sensitive probe-based detection methods that can detect target nucleic acids and proteins directly without amplification. The design uses multiple specific probes targeting different EV components simultaneously, achieving sufficient sensitivity through probe multiplicity rather than amplification cycles.
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
These biochips enable rapid, sensitive detection of EV contents and membrane proteins, reducing the number of sample preparation steps and improving detection sensitivity, allowing for early disease diagnosis from a single EV basis.
Implementation Method 1
molecular probes (MPs), such as molecular beacons and aptamers, to identify nucleic acids and protein content inside the captured EVs
Implementation Method 2
molecular probes (MPs), such as molecular beacons and aptamers, to identify nucleic acids and protein content inside the captured EVs
Implementation Method 3
antibody based capture system for extracellular vesicles (EVs)
Implementation Method 4
tethered cationic lipoplex nanoparticles (cTLNs) where intra-vesicular RNAs and DNAs are detected by the fusion of negatively charged EVs with positively charged lipoplex nanoparticles
Data Source
AI summary
The present invention disclosed a method of fabricating an antibody immunolipoplex nanoparticle (Ab-ILN) biochip and antibody tethered lipoplex nanoparticle (Ab-TLN) biochip. The aforementioned antibody-based lipoplex nanoparticle biochip or the related array contains molecular probes and is applied for detecting the presence of a disease or condition in a subject obtaining a body fluid sample by capturing and identifying both membrane protein and intra-vesicular DNA/RNA/proteins of extracellular vesicles (EVs).