Fluorescent Coating Evaluation for Protein Adsorbent Uniformity

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Solution Overview

Problem

It is challenging to confirm a uniform coating of protein adsorbents on complexly structured cell culture substrates using nondestructive tests, as existing methods are inadequate for evaluating the coating state on diversified and complicated substrate structures.

Innovation Solution

A method combining fluorescent dyes, protein adsorbents, and protein detection reagents is employed, where the substrate is irradiated with specific light to visualize the coating state by comparing fluorescence intensities in different configurations, allowing for nondestructive evaluation of protein adsorption and coating uniformity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If a nondestructive test is used to evaluate coating state, then the substrate structure can be preserved and reused, but the test cannot effectively visualize coating uniformity on complicated substrate structures

Engineering Contradiction:
Improvesubstrate reusabilityVSAvoidcoating visualization capability
Core Design Contradiction:
ReliabilityVSDifficulty of detecting and measuring

Solution Approach 1:

A fluorescent dye is introduced as an intermediary substance that binds to the protein adsorbent coating on the substrate. When excited by light, the dye emits fluorescence that can be detected and visualized, allowing nondestructive evaluation of coating uniformity on complex substrate structures while preserving the substrate for reuse

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The method utilizes fluorescence emission (a form of light/color change) from the dye molecules bound to the protein adsorbent. By irradiating the substrate with excitation light and detecting the emitted fluorescence, the coating state becomes visible without damaging the substrate, resolving the contradiction between nondestructive testing and visualization capability

Inventive Principle:
Principle #32Color changes

2Measurement precision

If a fluorescent dye is used to visualize coating state, then coating uniformity can be detected, but additional steps for dye application and excitation are required

Engineering Contradiction:
Improvecoating state detection accuracyVSAvoidevaluation process steps
Core Design Contradiction:
Measurement precisionVSDevice complexity

Solution Approach 1:

The fluorescent dye is combined with the protein adsorbent in the coating solution, so that when the coating is applied to the substrate, the dye is simultaneously deposited and bound to the protein adsorbent. This merging eliminates separate dye application steps and simplifies the overall evaluation process while maintaining precise coating state detection

Inventive Principle:
Principle #5Merging (Combining)

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 effective visualization and evaluation of protein adsorption states and coating uniformity on complex substrate structures, facilitating the assessment of protein adsorbent distribution and protein presence, even on intricate surfaces like hollow fiber bioreactors.

Implementation Method 1

a coating layer containing a protein adsorbent and a fluorescent dye is formed on at least one surface of a polymer substrate, the color development state 1 of the substrate 1 is detected by irradiating the substrate 1 with light

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 2

a coating layer containing a protein detection reagent is formed on the coating layer containing the protein adsorbent and the fluorescent dye, the color development state 2 of the substrate 2 is detected by irradiating the substrate 2 with light of the specific wavelength in which the protein detection reagent absorbs light

Methodology Applied
Scientific EffectLight absorption: Absorption (EM radiation)

Data Source

PatentEP4025899B1Method for evaluating a coating state of either an adsorbent of a sample protein or an adsorption state of a sample protein
Publication Date: 2023.11.08 TERUMO KK
  • EP4025899B1 patent drawingFigure 1A~1D
  • EP4025899B1 patent drawingFigure 2A~2D
  • EP4025899B1 patent drawingFigure 3

AI summary

A method for evaluating a coating state of either an adsorbent of a sample protein (40) or an adsorption state of the sample protein, the method comprising: providing a polymer substrate 1 (10) having a coating layer (20) containing the adsorbent of the sample protein and a fluorescent dye formed on at least one surface of the polymer substrate, and irradiating the substrate 1 with light to detect a color state 1 of the substrate 1; providing a polymer substrate 2 having a first coating layer (20) containing the adsorbent of the sample protein and a fluorescent dye and a second coating layer (30) containing a protein detection reagent (31) which conjugates with the sample protein, the first and second coating layers being sequentially formed on at least one surface of the polymer substrate, and irradiating the substrate 2 with light to detect a color state 2 of the substrate 2; providing a polymer substrate 3 having a coating layer containing the adsorbent of the sample protein and a fluorescent dye formed on at least one surface of the polymer substrate, introducing a sample containing the sample protein, which absorbs the irradiated light, onto the substrate 3 to obtain a substrate 4, and irradiating the substrate 4 with light to detect a color state 3 of the substrate 4; providing a polymer substrate 5 having a coating layer containing the adsorbent of the sample protein and a fluorescent dye formed on at least one surface of the polymer substrate, introducing a sample containing the sample protein (40) onto the substrate 5 to obtain a substrate 6, introducing the protein detection reagent (31) which has a higher degree of light absorption than that of the sample protein and which conjugates with the sample protein onto the polymer substrate 6 to obtain a substrate 7, and irradiating the substrate 7 with light to detect a color state 4 of the substrate 7;and comparing the color state 4 with the color states 1, 2 or 3 to evaluate a coating state of the adsorbent of the sample protein or the adsorption state of the sample protein.