Extracellular Vesicle Detection via Intrinsic Fluorescence

Solution Overview

Problem

Current methods for detecting and quantifying extracellular vesicles are laborious, time-consuming, and often produce heterogeneous mixes, lacking efficiency and reliability, especially in commercial applications.

Innovation Solution

The method involves detecting intrinsic fluorescence of extracellular vesicles using specific excitation and emission wavelengths without additional dyes or markers, employing techniques like column chromatography and filtration to isolate and quantify these vesicles accurately.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If ultra-centrifugation protocols are used for extracellular vesicle purification, then vesicles can be isolated, but the method produces heterogeneous mixes and requires laborious, time-consuming procedures

Engineering Contradiction:

Improvepurification reliabilityVSAvoidpurification time

Core Design Contradiction:

ReliabilityVSLoss of time

Solution Approach 1:

The patent replaces the mechanical ultra-centrifugation system with a fluorescence-based detection system. Instead of using centrifugal force to separate and identify vesicles, the invention uses intrinsic fluorescence properties to detect and quantify extracellular vesicles directly in the supernatant, eliminating the need for time-consuming centrifugation procedures while maintaining reliable identification of vesicles.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent exploits the intrinsic fluorescence property that naturally exists in extracellular vesicles without requiring any external labeling or preparation. The vesicles serve themselves by providing their own detection signal through their inherent fluorescent characteristics, eliminating the need for additional reagents or complex purification steps.

Inventive Principle:

Principle #25Self-service

2Measurement precision

If additional dyes or fluorophores are used to detect extracellular vesicles, then detection sensitivity can be improved, but the method complexity and cost increase

Engineering Contradiction:

Improvedetection sensitivityVSAvoiddetection method complexity

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The patent exploits the intrinsic fluorescence property that naturally exists in extracellular vesicles without requiring any external labeling or preparation. The vesicles serve themselves by providing their own detection signal through their inherent fluorescent characteristics, eliminating the need for additional reagents or complex purification steps.

Inventive Principle:

Principle #25Self-service

Solution Approach 2:

The patent extracts and utilizes only the essential intrinsic fluorescence property of extracellular vesicles for detection, removing the need for additional dyes, fluorophores, markers, or imaging compounds. This simplification maintains detection capability while reducing method complexity and cost.

Inventive Principle:

Principle #2Taking out (Extraction)

3Reliability

If current detection methods are used, then extracellular vesicles can be identified, but the methods lack efficiency and commercial reproducibility

Engineering Contradiction:

Improvedetection reliabilityVSAvoiddetection efficiency

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent replaces the mechanical ultra-centrifugation system with a fluorescence-based detection system. Instead of using centrifugal force to separate and identify vesicles, the invention uses intrinsic fluorescence properties to detect and quantify extracellular vesicles directly in the supernatant, eliminating the need for time-consuming centrifugation procedures while maintaining reliable identification of vesicles.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent enables continuous detection of extracellular vesicles by measuring intrinsic fluorescence in the supernatant without interruption for centrifugation or other batch processing steps. This continuous measurement approach improves productivity and ensures consistent, reproducible results suitable for commercial applications.

Inventive Principle:

Principle #20Continuity of useful action

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 enables rapid and reliable detection and quantification of extracellular vesicles, distinguishing them from contaminants and providing a stereotypical elution profile, thus improving upon existing methods in efficiency and purity.

Implementation Method 1

detecting intrinsic fluorescence of the extracellular vesicles without the use of additional dyes, fluorophores, markers, or imaging compounds

Methodology Applied

Scientific EffectIntrinsic fluorescence: Fluorescence

Data Source

PatentUS12158424B2Methods of measuring exosomes using intrinsic fluorescence

Publication Date: 2024.12.03 LONZA SALES AG

AI summary

Described herein are novel rapid and reliable methods of detection of extracellular vesicles and quantifying extracellular vesicle concentrations and absolute number from various sources, including raw cell harvest. The methods described herein comprise detection of intrinsic fluorescence of extracellular vesicles in biological samples. Extracellular vesicles analyzed by the methods of this application have a stereotypical elution profile distinct from known contaminants. The methods described herein are a significant improvement over the state of the art and fulfills an unmet need in the field of extracellular vesicle manufacturing and quality control.