Protamine-Coated Surfaces for Extracellular Vesicle Capture

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Solution Overview

Problem

Current methods for isolating extracellular vesicles (EVs) from biological fluids face challenges such as complexity, inefficiency, damage to vesicles during purification, and the need for large sample sizes, with existing techniques like ultracentrifugation and size exclusion chromatography being difficult to standardize and prone to contamination.

Innovation Solution

A method involving a solid surface coated with protamine hydrochloride, a polycationic substance, is used to capture and quantify EVs without additional reagents or processing steps, utilizing a microtiter plate, magnetic beads, or other solid surfaces to bind EVs based on their negative charge, allowing for efficient detection and quantification.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If differential ultracentrifugation or density gradient ultracentrifugation is used for EV purification, then EV isolation efficiency is improved, but EV integrity is damaged and membrane debris is observed

Engineering Contradiction:
ImproveEV isolation efficiencyVSAvoidEV integrity
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent replaces the mechanical ultracentrifugation system with a chemical adsorption system using protamine-coated surfaces. EVs are captured through electrostatic interactions between the positively charged protamine and negatively charged EV surfaces, eliminating mechanical shear stress while maintaining high capture efficiency

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the purification mechanism from mechanical force-based (ultracentrifugation) to electrostatic interaction-based (protamine adsorption). This parameter change allows EV isolation without the damaging high-speed rotation, preserving EV integrity while achieving efficient purification

Inventive Principle:
Principle #35Parameter changes

2Reliability

If size exclusion chromatography is used for EV purification, then EV integrity is maintained, but EV recovery efficiency is reduced

Engineering Contradiction:
ImproveEV integrityVSAvoidEV recovery efficiency
Core Design Contradiction:
ReliabilityVSProductivity

Solution Approach 1:

The patent replaces the mechanical chromatography system with a chemical adsorption system. Instead of relying on size-based physical separation through columns, EVs are captured through electrostatic adsorption to protamine-coated surfaces, achieving both high integrity and high recovery efficiency

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent uses a composite approach by coating solid surfaces with protamine, creating a material that combines the structural properties of the solid support with the electrostatic binding properties of protamine. This composite material enables efficient EV capture while maintaining integrity

Inventive Principle:
Principle #40Composite materials

3Quantity of substance

If polymeric precipitation methods are used for EV isolation, then EV concentration from small samples is improved, but EV contamination increases

Engineering Contradiction:
ImproveEV concentrationVSAvoidEV purity
Core Design Contradiction:
Quantity of substanceVSManufacturing precision

Solution Approach 1:

The patent applies local quality by using protamine coating specifically on the solid surface where EV capture is needed. This localized application ensures that only EVs are captured through electrostatic interactions, while other contaminants remain in solution, achieving both concentration and purity

Inventive Principle:
Principle #3Local quality

4Productivity

If conventional EV purification methods are used, then EV isolation is achieved, but the methods are complex and difficult to standardize

Engineering Contradiction:
ImproveEV isolation capabilityVSAvoidPurification method complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent extracts the essential function of EV purification from complex multi-step procedures and concentrates it into a single adsorption step using protamine-coated surfaces. This simplification maintains isolation capability while eliminating complexity and standardization difficulties

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent creates a universal platform using protamine-coated surfaces that can capture EVs from various biological fluids through a single standardized protocol. This multi-functional approach works across different sample types without requiring method optimization, reducing complexity

Inventive Principle:
Principle #6Universality (Multi-functionality)

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 approach simplifies the detection and quantification of EVs from small samples, maintaining their integrity and avoiding the need for expensive equipment, while providing a more efficient and effective method for diagnostic applications.

Implementation Method 1

A method involving a solid surface coated with protamine hydrochloride, a polycationic substance, is used to capture and quantify EVs without additional reagents or processing steps, utilizing a microtiter plate, magnetic beads, or other solid surfaces to bind EVs based on their negative charge

Methodology Applied
Scientific EffectElectrostatic attraction: Electrostatics

Data Source

PatentEP3458854B1Method and kit for capturing extracellular vesicles (EVS) on a solid surface
Publication Date: 2023.11.22 UNICYTE EV AG
  • EP3458854B1 patent drawingFigure 1
  • EP3458854B1 patent drawingFigure 2

AI summary

The invention relates to a method and a kit for capturing on a solid surface the extracellular vesicles (EVs) which are present in a biological fluid sample. The solid surface is coated with a polycationic substance, preferably a protamine salt such as protamine hydrochloride. The method and kit of the invention is useful for the detection and/or quantification of the extracellular vesicles (EVs) which are present in a biological fluid sample, particularly in a diagnostic context.