S100P Stabilization in Pancreatic Juice Using Protease Inhibitors

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Solution Overview

Problem

Existing methods fail to stably store S100P in pancreatic juice at room temperature due to protease activity, with previous solutions either ineffective for high protease samples or causing structural modifications to proteins during drying.

Innovation Solution

The use of sulfonyl fluoride group-containing protease inhibitors, such as PMSF and AEBSF, in combination with a trypsin-like serine protease inhibitor like TLCK, is employed to inhibit protease activity in pancreatic juice samples, allowing stable storage at room temperature.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Ease of operation

If pancreatic juice is stored at room temperature, then ease of storage is improved, but S100P is degraded by protease activity

Engineering Contradiction:
Improveease of storageVSAvoidstability of S100P
Core Design Contradiction:
Ease of operationVSStability of the object's composition

Solution Approach 1:

The protease inhibitors (PMSF, AEBSF, and TLCK) are added to the pancreatic juice sample immediately after collection, before storage. This preliminary action prevents protease activation and S100P degradation from the outset, allowing the sample to be stored at room temperature without compromising S100P stability.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

Protease inhibitors serve as intermediary substances that mediate between the proteases naturally present in pancreatic juice and the S100P protein. These inhibitors block protease activity, preventing direct interaction between proteases and S100P, thereby preserving S100P integrity during room temperature storage.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Stability of the object's composition

If protease inhibitors are added to inhibit degradation, then S100P stability is improved, but sample preparation complexity increases

Engineering Contradiction:
Improvestability of S100PVSAvoidcomplexity of sample preparation
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The kit provides protease inhibitors in advance, pre-packaged and ready for use. This allows clinicians to simply add the inhibitors to collected samples without needing to prepare complex inhibition systems, reducing the actual operational complexity despite the multi-component nature of the solution.

Inventive Principle:
Principle #10Preliminary action

Solution Approach 2:

The kit uses disposable, pre-packaged protease inhibitor formulations that are simple to add and require no complex handling or preparation. This approach trades the complexity of reusable, adjustable systems for simple, single-use packages that are easier to implement in clinical settings.

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

3Stability of the object's composition

If freezing is used to preserve S100P, then S100P stability is improved, but storage equipment requirements increase

Engineering Contradiction:
Improvestability of S100PVSAvoidadaptability to storage conditions
Core Design Contradiction:
Stability of the object's compositionVSAdaptability or versatility

Solution Approach 1:

The invention changes the critical parameter from temperature (requiring freezing) to chemical composition (adding protease inhibitors). This parameter change allows the same stability outcome to be achieved under different storage conditions, including room temperature, making the system adaptable to various storage environments without specialized equipment.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The protease inhibitor-based approach provides universal protection against protease-mediated degradation regardless of storage temperature. The same inhibitor formulation works whether samples are stored at room temperature, refrigerated, or frozen, making the system versatile and adaptable to different storage infrastructures.

Inventive Principle:
Principle #6Universality (Multi-functionality)

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

This method effectively suppresses degradation and modification of S100P, enabling high-quality sample preservation for clinical tests and contributing to the study and diagnosis of pancreatic cancer.

Implementation Method 1

adding, to a pancreatic juice component-containing biological sample, PMSF and AEBSF as a sulfonyl fluoride group-containing protease inhibitor and TLCK as a trypsin-like serine protease inhibitor belonging to a group of amino acid chloromethyl ketones

Methodology Applied
Scientific EffectProtease inhibition: Enzyme

Data Source

PatentEP2866029B1Method for preparing sample containing pancreatic juice component, and kit for storing biological sample containing pancreatic juice component at room temperature
Publication Date: 2017.05.03 OLYMPUS CORPORATION(JP)
  • EP2866029B1 patent drawingFigure 1~2
  • EP2866029B1 patent drawingFigure 3
  • EP2866029B1 patent drawingFigure 4

AI summary

The present invention provides a method for preparing a pancreatic juice component-containing sample capable of stably storing, particularly S100P, among proteins in a pancreatic juice component-containing biological sample at room temperature; and a kit suited for this method. The present invention is characterized in that a pancreatic juice component-containing sample to be stored at room temperature until it is provided for detection of S100P is prepared by adding, to a pancreatic juice component-containing biological sample, at least one sulfonyl fluoride group-containing protease inhibitor and at least one trypsin-like serine protease inhibitor belonging to a group of amino acid chloromethyl ketones.