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
Engineering 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
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.
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.
2Productivity
If a large amount of protease is used for albumin digestion, then digestion completeness is improved, but cost increases
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.
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.
3Productivity
If high concentration protease solution is prepared for adding large amount of protease, then digestion efficiency is improved, but protease autolysis increases
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.
4Ease of operation
If protease treatment is performed without denaturing agent, then procedure simplicity is maintained, but digestion efficiency is insufficient
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.
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
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
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
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
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
Implementation Method 6
the glycation amount of the albumin is obtained by measuring a chromogenic level of the chromogenic substrate
Data Source
Figure 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.