Dextrin-Based Protein Detection Reagents for Detergent Tolerance

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Solution Overview

Problem

Current protein detection and quantitation methods, particularly those using dye-based reagents like Coomassie Brilliant Blue, are significantly interfered with by detergents and surfactants, leading to instability and reduced sensitivity, making it difficult to accurately measure protein concentrations in samples containing these agents.

Innovation Solution

A reagent comprising a protein-complexing dye and dextrins, with dextrin concentrations between 0.01 to 200 mg/ml, is used to detect and quantify proteins, effectively trapping detergents in a dextrin-detergent complex and minimizing their interference, while maintaining protein-dye complex stability.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If dye-based reagents like Coomassie Brilliant Blue are used for protein detection, then sensitivity is improved, but interference from detergents and surfactants increases

Engineering Contradiction:
ImprovesensitivityVSAvoidinterference from detergents and surfactants
Core Design Contradiction:
Measurement precisionVSObject-affected harmful factors

Solution Approach 1:

The patent introduces dextrins as intermediary substances that selectively bind to detergents and surfactants, forming dextrin-detergent complexes. This mediator approach allows the protein-dye complex to remain unaffected by detergent interference while dextrins capture the harmful interfering substances, thereby resolving the contradiction between maintaining sensitivity and reducing interference

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent modifies the chemical composition parameters of the detection reagent by adding dextrins at specific concentrations (0.01 to 200 mg/ml). This parameter change alters the reagent's interaction profile, enabling it to tolerate the presence of detergents and surfactants while maintaining protein detection sensitivity

Inventive Principle:
Principle #35Parameter changes

2Ease of operation

If protein-dye complex is formed in detergent-rich environments, then protein detection is enabled, but stability of the complex deteriorates

Engineering Contradiction:
Improveprotein detection capabilityVSAvoidstability of protein-dye complex
Core Design Contradiction:
Ease of operationVSStability of the object's composition

Solution Approach 1:

Dextrins act as protective intermediaries that bind to detergents, preventing them from disrupting the protein-dye complex. This mediator mechanism allows the protein-dye complex to maintain its stability even in detergent-rich environments, enabling easy operation without compromising complex stability

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent applies prior cushioning by pre-forming dextrin-detergent complexes before they can interfere with the protein-dye complex formation. This preemptive binding of detergents by dextrins cushions the protein-dye complex against destabilizing effects, allowing stable detection in challenging environments

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

3Object-affected harmful factors

If extensive washing procedures are used to remove detergents, then interference is reduced, but time and complexity increase

Engineering Contradiction:
Improvedetergent interferenceVSAvoidwashing time
Core Design Contradiction:
Object-affected harmful factorsVSLoss of time

Solution Approach 1:

Instead of removing detergents through time-consuming washing, the patent uses dextrins as intermediaries to sequester detergents in situ. This approach eliminates the need for extensive washing procedures by chemically trapping detergents through dextrin-detergent complex formation, thereby reducing both time and operational complexity

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent extracts the harmful detergent interference from the detection system by having dextrins selectively bind and sequester detergents. This extraction approach removes detergent interference without requiring physical removal through washing, significantly reducing time and procedural complexity

Inventive Principle:
Principle #2Taking out (Extraction)

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 accurate and sensitive protein detection and quantitation even in detergent-rich environments, enhancing the stability of the protein-dye complex and reducing the need for extensive washing procedures, thus improving the reliability and efficiency of protein analysis.

Implementation Method 1

effectively trapping detergents in a dextrin-detergent complex

Methodology Applied
Scientific EffectComplex formation:

Implementation Method 2

a non-specific reaction in which a protein-complexing dye binds to the protein. The formation of a dye-protein complex causes a change in the optical properties of the dye

Methodology Applied
Scientific EffectDye-protein complex formation:

Implementation Method 3

there is a colour change proportional to the amount of protein present in the sample

Methodology Applied
Scientific EffectColorimetric detection: Absorption Spectroscopy

Data Source

PatentEP2295979B1Protein detection reagents and methods with dyes and dextrins
Publication Date: 2014.09.24 EXPEDON LTD
  • EP2295979B1 patent drawingFigure 1A~1B
  • EP2295979B1 patent drawingFigure 2A~2B
  • EP2295979B1 patent drawingFigure 3A~3B

AI summary

Protein Detection Reagents and methods with dyes and dextrins