Lactate Biosensor Using Binding Protein for Rapid Detection

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Solution Overview

Problem

Current lactate sensors suffer from long response times, short stability, and poor reproducibility, limiting their effectiveness in detecting lactate concentrations accurately and reliably across various applications.

Innovation Solution

Development of a biosensor using a reporter group attached to a lactate-binding protein that undergoes a signal change upon lactate binding, allowing for rapid, reliable, and accurate lactate detection without the need for additional substrates or enzymatic activity, with enhanced durability and applicability in clinical, food, and environmental settings.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If enzyme-based biosensors (lactate dehydrogenase, lactate oxidase, etc.) are used, then lactate detection capability is achieved, but response time is long and stability is short

Engineering Contradiction:
ImprovestabilityVSAvoidresponse time
Core Design Contradiction:
ReliabilityVSLoss of time

Solution Approach 1:

The patent extracts the enzymatic component from the biosensor system and replaces it with a direct binding protein-based detection mechanism. The lactate-binding protein directly binds lactate without requiring enzymatic catalysis, thereby eliminating the slow enzymatic reaction step and achieving rapid response time while maintaining stability through the use of stable protein structures.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent substitutes the biochemical enzymatic mechanism with a direct protein-ligand binding mechanism. Instead of using enzymes that catalyze chemical reactions, the invention uses lactate-binding proteins that directly bind lactate molecules, triggering conformational changes that can be detected optically or electrochemically without requiring substrate consumption or product generation.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

2Reliability

If enzyme-based biosensors are used, then lactate detection is possible, but reproducibility is poor

Engineering Contradiction:
ImprovereproducibilityVSAvoiddetection accuracy
Core Design Contradiction:
ReliabilityVSMeasurement precision

Solution Approach 1:

The patent changes the fundamental detection parameter from enzymatic reaction rates (which vary between preparations and over time) to direct binding affinity and conformational changes (which are more consistent and reproducible). The lactate-binding protein exhibits consistent binding characteristics that can be reliably reproduced across different sensor preparations, improving both reproducibility and measurement precision.

Inventive Principle:
Principle #35Parameter changes

3Ease of manufacture

If additional substrates are required for signal development, then enzymatic activity can be maintained, but device complexity and cost increase

Engineering Contradiction:
Improvemanufacturing simplicityVSAvoidreagent requirements
Core Design Contradiction:
Ease of manufactureVSDevice complexity

Solution Approach 1:

The patent implements a self-service detection system where the lactate-binding protein itself serves as both the recognition element and the signaling mechanism. The protein undergoes conformational changes upon lactate binding that directly trigger signal generation without requiring external substrates, cofactors, or additional reagents. This eliminates the need for complex substrate delivery systems and reduces both device complexity and manufacturing costs.

Inventive Principle:
Principle #25Self-service

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 provides rapid and accurate lactate detection, maintaining functionality over extended periods and in diverse conditions, enabling continuous monitoring and reducing costs through simplified manufacturing and distribution processes.

Implementation Method 1

The lactate-binding protein includes a domain or region(s) that binds the lactate. The domain or region involved in ligand binding is comprised of a plurality of residues, e.g., non-contiguous amino acids of the ligand-binding protein, which are contact points or sites of contact between the ligand and its cognate ligand-binding protein.

Methodology Applied
Scientific EffectLigand binding:

Implementation Method 2

The binding of a lactate to the lactate-binding domain of the lactate-binding protein causes a change in signaling by the reporter group. A reporter group that transduces a detectable signal may be attached to the lactate-binding proteins (biosensors) described herein.

Methodology Applied
Scientific EffectSignal transduction:

Implementation Method 3

detectable signal comprises a fluorescent, electrochemical, nuclear magnetic resonance (NMR), or electron paramagnetic resonance (EPR) signal

Methodology Applied
Scientific EffectFluorescence: Fluorescence

Implementation Method 4

detectable signal comprises a fluorescent, electrochemical, nuclear magnetic resonance (NMR), or electron paramagnetic resonance (EPR) signal

Methodology Applied
Scientific EffectElectrochemical signal transduction:

Data Source

PatentEP3377897B1Lactate biosensors and uses thereof
Publication Date: 2024.05.01 DUKE UNIV
  • EP3377897B1 patent drawingFigure 1A~1D
  • EP3377897B1 patent drawingFigure 2A~2D
  • EP3377897B1 patent drawingFigure 3A~3B

AI summary

The preseni subject matter provides lactate biosensors as well as compositions, devices, and methods comprising such biosensors.