FADGDH Purification for Glucose Sensor Accuracy
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Glucose sensors using glucose-measuring enzyme preparations often produce inaccurate measurements due to factors like trehalose contamination and insufficient purification of flavin adenine dinucleotide-dependent glucose dehydrogenase (FADGDH), leading to higher calculated glucose concentrations or blank-up issues.
Innovation Solution
A composition comprising FADGDH with low trehalase content and minimal trehalose degradation activity, achieved through purification processes such as chromatography, is developed to minimize contamination and enhance measurement accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If FADGDH is purified from original organisms or host organisms, then the enzyme preparation can be obtained, but contaminating proteins including trehalase may be present causing measurement errors
Solution Approach 1:
The patent applies the extraction principle by specifically removing trehalase and other contaminating proteins from the FADGDH preparation through purification processes. This is achieved by extracting only the desired FADGDH enzyme while leaving contaminating proteins behind, thereby obtaining a high-purity enzyme preparation that prevents false glucose measurements caused by trehalose degradation.
Solution Approach 2:
The patent employs parameter changes by optimizing purification conditions such as pH, temperature, and chromatography parameters to maximize the separation between FADGDH and contaminating proteins. By adjusting these parameters, the purification process effectively removes trehalase while maintaining FADGDH activity, resolving the contradiction between obtaining sufficient enzyme quantity and ensuring measurement reliability.
2Ease of manufacture
If trehalase is present in FADGDH preparation, then the preparation can be produced, but trehalose is degraded to produce glucose leading to false high measured values
Solution Approach 1:
The patent converts the harmful effect of trehalase into a beneficial purification target. By identifying trehalase as the source of measurement errors, the invention designs purification processes specifically aimed at removing this contaminant. The harm caused by trehalase presence is transformed into a clear criterion for purification, enabling the production of high-quality FADGDH preparations that provide accurate glucose measurements.
3Productivity
If insufficient purification is performed, then production cost and time are reduced, but contaminating proteins cause defects in glucose sensors
Solution Approach 1:
The patent applies preliminary action by performing purification steps before the FADGDH preparation is used in glucose sensor production. This advance purification removes contaminating proteins including trehalase, preventing potential sensor defects before they occur. By addressing the purification issue in advance, the invention ensures both sensor reliability and production efficiency without compromising either aspect.
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 solution provides a glucose dehydrogenase composition that accurately measures glucose concentrations by reducing the risk of false high values and blank-up, resulting in highly reliable glucose measurement kits and sensors.
Implementation Method 1
purification processes such as chromatography
Implementation Method 2
flavin adenine dinucleotide-dependent glucose dehydrogenase (FADGDH)
Data Source
AI summary
Provided is GDH with increased applicability to glucose sensors. A composition contains FADGDH, wherein when 0.1 mL of the composition is added to 2.9 mL of a solution containing 10 mM of trehalose and 1 mmol/L of potassium ferricyanide to give a glucose dehydrogenase activity of 500 U/mL and incubated at 37° C., the decrease in absorbance at 405 nm resulting from reduction of the potassium ferricyanide is less than 20 mAbs per minute.