Xanthene Dye Fluorescence for Protein Aggregation Detection

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Solution Overview

Problem

Current methods for evaluating the aggregation state of proteins in solutions have limited dynamic range, making it difficult to identify time-dependent changes effectively.

Innovation Solution

Utilizing light-absorption/fluorescence characteristics of xanthene dyes, such as rose bengal, eosin Y, or erythrosin B, to assess the aggregation state of proteins by measuring changes in fluorescence spectrum, intensity, or absorbance over time.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If turbidity of the solution is used as an index to evaluate aggregation state, then the evaluation method is simple, but the dynamic range is small and only part of time-dependent changes can be identified

Engineering Contradiction:

Improveevaluation method simplicityVSAvoiddynamic range of aggregation state changes

Core Design Contradiction:

Ease of operationVSLoss of information

Solution Approach 1:

The patent changes the measurement parameter from turbidity (light scattering) to fluorescence intensity of xanthene dye. This parameter change enables detection across a broader dynamic range of aggregation states, capturing time-dependent changes from monomer to fibril formation that turbidity cannot detect

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent replaces the mechanical/optical measurement of turbidity with a chemical/optical measurement using fluorescence spectroscopy. The xanthene dye binds to aggregated proteins and exhibits enhanced fluorescence, providing a more sensitive and broader-range detection method

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

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 allows for a more comprehensive evaluation of protein aggregation states, enabling the identification of multiple stages of aggregation from monomers to fibrils, and can be used for screening drugs or isolating protein aggregates.

Implementation Method 1

light-absorption/fluorescence characteristic(s) of a xanthene dye

Methodology Applied

Scientific EffectFluorescence: Fluorescence

Implementation Method 2

light-absorption/fluorescence characteristic(s) of a xanthene dye

Methodology Applied

Scientific EffectLight absorption: Absorption (EM radiation)

Data Source

PatentUS20240319202A1Method for evaluating aggregation state of aggregative protein

Publication Date: 2024.09.26 HAMAMATSU PHOTONICS KK

AI summary

A method for evaluating an aggregation state of aggregative proteins in a solution is disclosed, the method comprising evaluating an aggregation state of aggregative proteins using, as an index, light-absorption/fluorescence characteristic(s) of a certain xanthene dye in a solution comprising the aggregative proteins and the xanthene dye or a salt thereof.