RGO Biosensor for L-Glutamate Detection at Low Voltage

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Solution Overview

Problem

Existing L-glutamate biosensors have limitations such as short lifetime, high operating voltage, and narrow linear detection range, which restrict their accuracy and reliability in detecting L-glutamate concentrations in biological samples.

Innovation Solution

The development of an oxidizing enzyme- and thermally-reduced-graphene-oxide-(RGO)-based amperometric biosensor that operates at a reduced potential of -0.1 V, utilizing L-glutamate oxidase immobilized in an albumin and RGO matrix, to enhance sensitivity and stability while reducing interference from other compounds.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If conventional L-glutamate biosensors use Pt electrodes and GluOx enzyme, then sensitivity and response time are improved, but operating voltage increases to 0.5-0.7 V and linear detection range becomes narrow

Engineering Contradiction:
ImprovesensitivityVSAvoidoperating voltage
Core Design Contradiction:
Measurement precisionVSUse of energy by moving object

Solution Approach 1:

The patent changes the electrode material from Pt to RGO (reduced graphene oxide) and modifies the enzyme immobilization matrix by adding TiO2 nanoparticles. This parameter change in materials composition enables the biosensor to operate at lower voltage (-0.1 V) while maintaining sensitivity through enhanced electron transfer properties of RGO and TiO2.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite structure combining RGO, TiO2 nanoparticles, and GluOx enzyme in an albumin matrix. This composite material synergistically improves electron transfer (RGO), provides catalytic activity (TiO2), ensures enzyme stability (albumin), and maintains high sensitivity while enabling low operating voltage.

Inventive Principle:
Principle #40Composite materials

2Speed

If conventional L-glutamate biosensors use Pt electrodes and GluOx enzyme, then response time is improved, but lifetime becomes short

Engineering Contradiction:
Improveresponse timeVSAvoidlifetime
Core Design Contradiction:
SpeedVSDuration of action of moving object

Solution Approach 1:

The patent employs a disposable amperometric biosensor design where the RGO-TiO2-GluOx composite membrane is prepared on a inexpensive support. The biosensor is designed for single-use or limited-use applications, eliminating the need for complex regeneration procedures and extending practical operational lifetime while maintaining fast response characteristics.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent changes the enzyme immobilization matrix by incorporating TiO2 nanoparticles into the albumin-RGO composite. This parameter change in the matrix composition enhances enzyme stability and prevents leaching, thereby extending the biosensor's operational lifetime while preserving fast response time through efficient electron transfer pathways.

Inventive Principle:
Principle #35Parameter changes

3Productivity

If conventional L-glutamate biosensors are designed for in vivo use, then real-time detection is improved, but operational stability deteriorates with sensitivity decrease of 46.32% after 9 hours

Engineering Contradiction:
Improvereal-time detectionVSAvoidoperational stability
Core Design Contradiction:
ProductivityVSReliability

Solution Approach 1:

The patent extracts the biosensor from in vivo application and designs it for in vitro use with direct sample application. This removes the sensor from the complex in vivo environment that causes instability, while maintaining real-time detection capability through direct amperometric measurement of L-glutamate in extracted biological samples.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent provides beforehand cushioning for operational stability by incorporating TiO2 nanoparticles and albumin in the enzyme immobilization matrix. This stable composite structure protects the GluOx enzyme from denaturation and leaching, ensuring consistent sensitivity over extended measurement periods suitable for in vitro investigations.

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

4Measurement precision

If conventional L-glutamate biosensors are used, then detection capability is improved, but linear detection range becomes narrow, failing to cover the 5-100 μM range in plasma

Engineering Contradiction:
Improvedetection capabilityVSAvoidlinear detection range
Core Design Contradiction:
Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The patent changes the electrode material from Pt to RGO and modifies the enzyme immobilization matrix by adding TiO2 nanoparticles. This parameter change in materials composition broadens the linear detection range to cover 5-100 μM concentrations in plasma by optimizing electron transfer efficiency and enzyme activity across a wider concentration range.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite structure of RGO-TiO2-albumin-GluOx that synergistically broadens the linear detection range. RGO provides extensive surface area and efficient electron transfer, TiO2 enhances catalytic activity, and albumin ensures uniform enzyme distribution, collectively enabling accurate detection across the 5-100 μM range required for plasma analysis.

Inventive Principle:
Principle #40Composite materials

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 biosensor achieves a wide analytical range, improved stability, and reduced operating voltage, enabling accurate detection of L-glutamate in various biofluids, including serum and brain extracts, with high sensitivity and minimal interference from other neurochemicals.

Implementation Method 1

L-glutamate oxidase (GluOx) and Pt or Pt-modified electrodes

Methodology Applied
Scientific EffectEnzyme catalysis: Enzyme

Implementation Method 2

oxidizing enzyme- and thermally-reduced-graphene-oxide-(RGO)-based amperometric biosensor for direct evaluation of L-glutamate

Methodology Applied
Scientific EffectOxidation: Oxidation

Implementation Method 3

thermally-reduced-graphene-oxide-(RGO)-based amperometric biosensor... operates at a reduced potential of -0.1 V

Methodology Applied
Scientific EffectElectron transfer: Conduction (electrical)

Implementation Method 4

thermally-reduced-graphene-oxide-(RGO)

Methodology Applied
Scientific EffectThermal reduction: Thermolysis

Implementation Method 5

amperometric biosensor for direct evaluation of L-glutamate in biological media

Methodology Applied
Scientific EffectAmperometric detection:

Data Source

PatentEP4542212A1Electrochemical biosensor of metabolites for in vitro investigations in biological samples
Publication Date: 2025.04.23 VILNIUS UNIV
  • EP4542212A1 patent drawingFigure 1~2
  • EP4542212A1 patent drawingFigure 3~4B
  • EP4542212A1 patent drawing

AI summary

The invention discloses an enzyme- and carbonaceous material (e.g. thermallyreduced graphene oxide (RGO))-based amperometric biosensor for direct evaluation of selected metabolites in biological media such as brain extract or serum. The biosensor comprises a working electrode (2), and a permeable membrane (1) covered with immobilized metabolite-selective enzyme with the carbonaceous material, being in contact with the electrode (2) surface, for bioelectrocatalytic oxidation of the metabolite, thus measuring its presence. The determination of a specific metabolite - L-glutamate - is based on registration at -0.1 V (vs. Ag/AgCl) of anodic current generated by the electrooxidation of ammonia released during the reaction catalyzed by L-glutamate oxidase, while L-glutamate is converting to alfa-ketoglutarate. The biosensor comprises a working graphite electrode (2) and adjustable semipermeable membrane (1) comprising L-glutamate oxidase (EC 1.4.3.11) immobilized into albumin and thermally reducated graphene oxide (RGO).