Pyrophosphate Sensor with Segmented Reagent Layers

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Solution Overview

Problem

Conventional methods for measuring pyrophosphate and SNP typing face challenges in achieving stable characteristics due to the separate arrangement of buffer components and enzymes in dried reagent forms, leading to instability and reduced sensitivity.

Innovation Solution

A sensor system with an insulative substrate featuring a layered configuration of reaction reagent layers, including pyrophosphatase, glyceraldehyde-3-phosphate dehydrogenase, diaphorase, oxidized nicotineamide adenine dinucleotide, and a buffer component, separated to maintain enzyme stability and enhance sensitivity, allowing for high-sensitivity pyrophosphate detection and SNP typing without the need for large optical systems.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Device complexity

If buffer components and enzymes are arranged separately in dried reagent forms, then the sensor structure is simplified, but measurement stability and sensitivity deteriorate

Engineering Contradiction:

Improvesensor structureVSAvoidmeasurement stability

Core Design Contradiction:

Device complexityVSReliability

Solution Approach 1:

The sensor is divided into distinct functional layers: a buffer layer containing buffer components and a separate enzyme layer containing enzymes. This segmentation allows each layer to be optimized independently while maintaining overall measurement stability through proper spatial arrangement.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The enzyme layer is positioned directly on top of the buffer layer, creating a nested structure where the enzyme layer utilizes the buffer layer's properties while adding its own functional capability. This nested arrangement maintains stability by ensuring proper interaction between layers.

Inventive Principle:

Principle #7Nested doll (Nesting)

2Measurement precision

If conventional optical measurement systems are used for pyrophosphate detection, then measurement capability is achieved, but device size and complexity increase

Engineering Contradiction:

Improvepyrophosphate detection capabilityVSAvoidoptical system size

Core Design Contradiction:

Measurement precisionVSDevice complexity

Solution Approach 1:

The conventional optical measurement system is replaced with an electrochemical detection system using electrodes. This substitution eliminates the need for large optical components while maintaining pyrophosphate detection capability through electrochemical reactions that produce measurable electrical signals.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Measurement precision

If concentrated sulfuric acid is used in chemical methods for pyrophosphate measurement, then measurement is achieved, but safety and ease of operation worsen

Engineering Contradiction:

Improvepyrophosphate measurement capabilityVSAvoidsafety and convenience

Core Design Contradiction:

Measurement precisionVSEase of operation

Solution Approach 1:

The sensor uses a disposable design with dried reagent layers that are pre-prepared and stable. This eliminates the need for handling hazardous chemicals like concentrated sulfuric acid during operation, as the reagents are already in a stable, non-hazardous dried form on the sensor chip.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

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 sensor system provides stable and high-sensitivity pyrophosphate detection and SNP typing, with improved linearity and proportionality between pyrophosphate concentration and electric current, enabling efficient and convenient measurement.

Implementation Method 1

pyrophosphate is hydrolyzed into phosphoric acid by pyrophosphatase

Methodology Applied

Scientific EffectHydrolysis: Hydrolysis

Implementation Method 2

glyceraldehyde-3-phosphate dehydrogenase, diaphorase, glyceraldehyde-3-phosphate, oxidized nicotineamide adenine dinucleotide

Methodology Applied

Scientific EffectOxidation: Oxidation

Implementation Method 3

diaphorase, glyceraldehyde-3-phosphate, oxidized nicotineamide adenine dinucleotide, an electronic mediator

Methodology Applied

Scientific EffectElectron transfer: Redox Reactions

Data Source

PatentUS7632392B2Sensor of pyrophosphate and SNP typing sensor using the same

Publication Date: 2009.12.15 PANASONIC HOLDINGS CORP

AI summary

A sensor of pyrophosphate which can detect pyrophosphate conveniently with high sensitivity in a method for measuring pyrophosphate in SNP typing utilizing a primer extension reaction is provided.A sensor of pyrophosphate which is characterized by including: an insulative substrate 1; an electrode system that is formed thereon and has a measurement electrode 2 and a counter electrode 3; and a plurality of reaction reagent layers that are provided on the substrate 1 and include pyrophosphatase, glyceraldehyde-3-phosphate dehydrogenase, diaphorase, glyceraldehyde-3-phosphate, oxidized nicotineamide adenine dinucleotide, an electronic mediator, a magnesium salt and a buffer component, reaction reagent layer 36 including the enzyme being separated from reaction reagent layer 35 including the buffer component, and reaction reagent layer 37 including glyceraldehyde-3-phosphate being separated from the reaction reagent layer 35 including the buffer component.