Albumin Denaturing Agent for Efficient Protease Digestion

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Solution Overview

Problem

Conventional methods for assaying glycated albumin in blood samples require a large amount of protease, leading to enzyme deactivation, stability issues, and high costs, making efficient albumin digestion challenging.

Innovation Solution

An albumin denaturing agent containing quaternary ammonium compounds or coconut amine acetate is used to facilitate efficient protease treatment, allowing for sufficient albumin digestion with a small amount of protease.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If a large amount of protease is added to digest albumin in blood samples, then digestion efficiency is improved, but enzyme deactivation and stability issues occur

Engineering Contradiction:
Improvealbumin digestion efficiencyVSAvoidprotease stability
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent introduces a denaturing agent as an intermediary substance that modifies albumin structure to make it more accessible to protease. This mediator enables efficient digestion without requiring excessive protease, thereby maintaining protease stability and preventing deactivation while achieving complete albumin digestion.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent changes the physical-chemical parameters of albumin by adding a denaturing agent, which alters the conformation and accessibility of peptide bonds. This parameter change allows protease to work more efficiently at lower concentrations, improving both digestion efficiency and protease stability.

Inventive Principle:
Principle #35Parameter changes

2Productivity

If a large amount of protease is used for albumin digestion, then digestion completeness is improved, but cost increases

Engineering Contradiction:
Improvedigestion completenessVSAvoidprotease amount
Core Design Contradiction:
ProductivityVSQuantity of substance

Solution Approach 1:

The denaturing agent serves as a mediator that prepares albumin for protease action, enabling complete digestion with minimal protease. This intermediary function reduces the quantity of protease required while maintaining complete digestion of albumin in blood samples.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

By changing the conformational parameters of albumin through denaturing agent addition, the patent makes peptide bonds more accessible to protease. This parameter modification allows complete digestion to be achieved with a small amount of protease, reducing substance consumption.

Inventive Principle:
Principle #35Parameter changes

3Productivity

If high concentration protease solution is prepared for adding large amount of protease, then digestion efficiency is improved, but protease autolysis increases

Engineering Contradiction:
Improvedigestion efficiencyVSAvoidprotease composition stability
Core Design Contradiction:
ProductivityVSStability of the object's composition

Solution Approach 1:

The denaturing agent acts as a mediator that enhances the interaction between protease and albumin, allowing efficient digestion at lower protease concentrations. This prevents protease autolysis that would occur in high concentration solutions while maintaining high digestion efficiency.

Inventive Principle:
Principle #24Intermediary (Mediator)

4Ease of operation

If protease treatment is performed without denaturing agent, then procedure simplicity is maintained, but digestion efficiency is insufficient

Engineering Contradiction:
Improveprocedure simplicityVSAvoiddigestion efficiency
Core Design Contradiction:
Ease of operationVSProductivity

Solution Approach 1:

The patent introduces a denaturing agent as a simple intermediary step that dramatically improves digestion efficiency. This single addition enhances peptide bond accessibility without complicating the overall procedure, making the method both simple and highly effective.

Inventive Principle:
Principle #24Intermediary (Mediator)

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

The albumin denaturing agent enables efficient albumin digestion and glycated albumin assay, reducing the need for large protease amounts, improving assay efficiency, and addressing stability and cost issues in the medical field.

Implementation Method 1

an albumin denaturing agent for digesting an albumin by a protease efficiently... Since an albumin can be denatured by using the albumin denaturing agent of the present invention

Methodology Applied
Scientific EffectProtein denaturation:

Implementation Method 2

a step of digesting the albumin in the sample by a protease... protease treatment is performed with respect to the sample in the presence of the albumin denaturing agent

Methodology Applied
Scientific EffectEnzymatic hydrolysis: Enzyme

Implementation Method 3

a fructosyl amino acid oxidase (hereinafter, referred to as a 'FAOD') is allowed to act on a glycated part of the albumin, thereby generating hydrogen peroxide

Methodology Applied
Scientific EffectEnzymatic oxidation: Enzyme

Implementation Method 4

allowing a glycated part of the obtained albumin digestion product to react with FAOD... an amount of this hydrogen peroxide corresponds to a glycation amount of the albumin

Methodology Applied
Scientific EffectHydrogen peroxide generation: Hydrogen Peroxide

Implementation Method 5

a peroxidase (hereinafter, referred to as a 'POD') and a chromogenic substrate that develops a color by oxidation are added to this reaction liquid, so as to effect a redox reaction between the hydrogen peroxide and the chromogenic substrate

Methodology Applied
Scientific EffectRedox reaction: Redox Reactions

Implementation Method 6

the glycation amount of the albumin is obtained by measuring a chromogenic level of the chromogenic substrate

Methodology Applied
Scientific EffectColor development:

Data Source

PatentEP1950309B1Albumin-denaturing agent
Publication Date: 2011.08.17 ARKRAY INC
  • EP1950309B1 patent drawingFigure 1

AI summary

An albumin denaturing agent for digesting an albumin by a protease efficiently is provided. The albumin denaturing agent contains quaternary ammonium having a hydrocarbon group with a carbon number of 12 or more, or a salt of the quaternary ammonium. The albumin in a sample is digested by the protease in the presence of the albumin denaturing agent, a glycated part of the thus obtained albumin digestion product and a FAOD effect a reaction, and a redox reaction between the glycated part and the FAOD is measured, thereby determining a ratio (GA (%)) of the glycated albumin of the glycated albumin with respect to the albumin.