Peroxidase Reaction Accelerating Agent for Diagnostic Sensitivity
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Solution Overview
Problem
The yield of horseradish peroxidase (HRP) is influenced by weather factors and recombinant expression methods result in low expression levels and altered sugar chains, affecting antibody labeling in ELISA, necessitating a method to effectively utilize and accelerate peroxidase reactions.
Innovation Solution
A peroxidase reaction accelerating agent represented by the formula R1—N═N—R2, where R1 and R2 are aromatic rings optionally substituted with various groups, is used to enhance the enzymatic reaction catalyzed by peroxidase, allowing for more efficient hydrogen peroxide decomposition.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If HRP is obtained by harvesting cultivated Armoracia rusticana, then the yield is stable, but the production is influenced by weather factors such as drought or change in temperature
Solution Approach 1:
The patent introduces a peroxidase reaction accelerating agent as an intermediary substance that enhances the catalytic activity of peroxidase. This agent mediates between the peroxidase enzyme and the substrate, accelerating the reaction without being consumed, thereby improving productivity while maintaining reliability of natural HRP production
Solution Approach 2:
The patent changes the reaction parameters by introducing a compound that modifies the reaction kinetics. The accelerating agent alters the reaction rate parameter, allowing faster decomposition of hydrogen peroxide with the same amount of peroxidase, thus improving productivity without affecting the stability of natural production
2Reliability
If recombinant expression is used to produce peroxidase, then the yield can be stabilized, but the expression level is low and sugar chains are added affecting antibody labeling
Solution Approach 1:
The peroxidase reaction accelerating agent serves as an intermediary that compensates for the low expression level of recombinant peroxidase. By enhancing the catalytic efficiency, the same amount of recombinant peroxidase can achieve the desired reaction rate, making it unnecessary to produce large amounts of recombinant protein that would require complex purification to remove sugar chains
Solution Approach 2:
Instead of attempting to create a perfect recombinant peroxidase copy that mimics natural HRP exactly, the patent uses a simpler recombinant version with added sugar chains and compensates for its reduced efficiency through the accelerating agent. This copying approach with compensation is more feasible than creating an identical replica
3Productivity
If large amounts of peroxidase are used in recombinant expression systems, then the reaction rate can be maintained, but the cost and complexity of production increase
Solution Approach 1:
The patent changes the kinetic parameters of the reaction by introducing the accelerating agent. This allows maintaining a high reaction rate with a smaller amount of peroxidase, thereby simplifying the production system and reducing the complexity associated with producing and handling large amounts of enzyme
Solution Approach 2:
The accelerating agent acts as a mediator that amplifies the effect of peroxidase, allowing a small amount of enzyme to catalyze the decomposition of a large amount of hydrogen peroxide. This intermediary approach reduces the need for complex production facilities capable of handling large-scale enzyme production
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 peroxidase reaction accelerating agent accelerates hydrogen peroxide decomposition, reducing the amount of peroxidase needed in diagnostic reagents, increasing measurement sensitivity, and shortening reaction times.
Implementation Method 1
a peroxidase reaction accelerating agent which accelerates a peroxidase reaction
Data Source
AI summary
Provided is a compound capable of accelerating the enzymatic reaction catalyzed by peroxidase. The present invention provides a peroxidase reaction accelerating agent comprising a compound represented by formula (I) and a method for measuring hydrogen peroxide using the same.