Glutamate Dehydrogenase Stabilization in Aqueous Solutions

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Solution Overview

Problem

Glutamate dehydrogenase (GDH) from Clostridium bacteria loses its antigenic properties when dissolved in aqueous solutions, leading to short expiration times and increased risks of error in diagnostic tests, as existing stabilization methods do not effectively preserve its structure and function in such environments.

Innovation Solution

Incorporating a carboxylic acid with a carbon chain of at least 3 atoms and 2 -COOH groups or its salts into the aqueous solution containing GDH, which stabilizes the protein's three-dimensional structure and maintains its antigenic properties, allowing for extended storage and use in diagnostic assays.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If GDH is dissolved in aqueous solution for diagnostic tests, then the protein becomes accessible for detection assays, but it loses its antigenic properties and degrades over time

Engineering Contradiction:
ImproveAccessibility for detection assaysVSAvoidAntigenic properties retention
Core Design Contradiction:
Ease of operationVSReliability

Solution Approach 1:

The patent introduces a stabilizing compound as an intermediary substance that mediates between the aqueous solution environment and the GDH protein. This compound prevents direct harmful interactions between the aqueous environment and the protein's antigenic structures, allowing the protein to maintain its functional properties while remaining dissolved and accessible for diagnostic assays.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent modifies the chemical parameters of the aqueous solution by adding specific stabilizing compounds. This changes the solution's composition to create a more favorable environment for GDH stability, preventing degradation while maintaining solubility and accessibility for detection purposes.

Inventive Principle:
Principle #35Parameter changes

2Ease of manufacture

If GDH is stored in aqueous solution, then it remains ready for immediate use in tests, but the shelf life is significantly reduced

Engineering Contradiction:
ImproveReadiness for immediate useVSAvoidShelf life
Core Design Contradiction:
Ease of manufactureVSDuration of action of stationary object

Solution Approach 1:

The patent applies preliminary stabilization action by adding stabilizing compounds to the aqueous GDH solution during preparation. This preliminary protective measure ensures that the protein maintains its stability throughout storage, extending the shelf life while keeping the solution ready for immediate use without requiring additional preparation steps.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The stabilizing compound acts as a protective intermediary that preserves the protein's integrity during storage. This intermediary substance creates a stable environment that prevents degradation over time, thereby extending shelf life while maintaining the solution's readiness for immediate diagnostic use.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Stability of the object's composition

If existing stabilization methods are used for GDH in aqueous solution, then some structural preservation is achieved, but antigenic properties are not effectively maintained

Engineering Contradiction:
ImproveStructural preservationVSAvoidAntigenic properties maintenance
Core Design Contradiction:
Stability of the object's compositionVSReliability

Solution Approach 1:

The patent identifies and applies specific parameter changes in the solution composition, particularly the addition of particular stabilizing compounds at optimized concentrations. These parameter changes address the insufficiency of existing methods by creating conditions that specifically preserve antigenic properties rather than just general structural stability.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent introduces a specific stabilizing compound as an intermediary that selectively protects antigenic determinants. This intermediary substance appears to have a specific interaction mechanism that preserves the conformational structures responsible for antigenic properties, which existing methods fail to maintain effectively.

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 stabilization of GDH using carboxylic acids significantly extends the shelf life of aqueous solutions, ensuring the protein retains its antigenic properties and binding capabilities, enhancing the reliability and duration of diagnostic tests for detecting Clostridium bacteria.

Implementation Method 1

Incorporating a carboxylic acid with a carbon chain of at least 3 atoms and 2 -COOH groups or its salts into the aqueous solution containing GDH, which stabilizes the protein's three-dimensional structure and maintains its antigenic properties

Methodology Applied
Scientific EffectProtein stabilization:

Data Source

PatentEP3080144B1Stabilization of glutamate dehydrogenase in an aqueous solution
Publication Date: 2021.07.14 BIOMERIEUX SA
  • EP3080144B1 patent drawingFigure 1
  • EP3080144B1 patent drawingFigure 2
  • EP3080144B1 patent drawing

AI summary

The present invention relates to a process for stabilizing glutamate dehydrogenase from a bacterium of the Clostridium genus, in an aqueous solution, in order to maintain the antigenic properties thereof, comprising the step of mixing said glutamate dehydrogenase and a stabilizing composition which is a carboxylic acid having a carbon-based chain of at least 3 carbon atoms and comprising at least 2 -COOH groups, or a salt thereof. It also relates to the GDH compositions thus stabilized, and to the use thereof for detecting the presence of bacteria of the Clostridium genus.