Polyol Stabilization of Biological Samples for Molecular Analysis

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Solution Overview

Problem

Current methods for stabilizing biological samples, particularly nucleic acids and proteins, are inadequate for molecular biological analysis, as they often lead to degradation or alteration of these molecules, and existing reagents are either ineffective for compact tissue samples or pose safety and logistical challenges, such as the use of toxic or flammable substances and the need for extreme cold storage.

Innovation Solution

A method involving the use of polyol compositions, specifically polyethylene glycols and polypropylene glycols, to stabilize biological samples at moderate temperatures (0°C to 80°C), allowing for both histological and molecular biological analysis without the need for cooling devices or hazardous substances, effectively preserving nucleic acids and proteins for extended periods.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Stability of the object's composition

If formalin and paraffin embedding are used to stabilize biological samples, then histological examination is enabled, but molecular biological analysis is compromised due to inadequate nucleic acid stabilization

Engineering Contradiction:
Improvenucleic acid stabilityVSAvoidmolecular biological analysis reliability
Core Design Contradiction:
Stability of the object's compositionVSReliability

Solution Approach 1:

The patent changes the chemical parameters of the stabilization medium by using liquid nitrogen at -70°C instead of formalin, fundamentally altering the stabilization mechanism from chemical crosslinking to physical freezing, which preserves nucleic acids for molecular analysis while maintaining histological integrity

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent creates a composite stabilization system combining liquid nitrogen freezing with specific buffer compositions (containing glycine, Tris, EDTA, and表面活性 agents) that work synergistically to stabilize both nucleic acids and proteins simultaneously

Inventive Principle:
Principle #40Composite materials

2Stability of the object's composition

If crosslinking stabilizing agents like formalin are used, then tissue structure is preserved, but nucleic acid extractability is reduced

Engineering Contradiction:
Improvetissue structure stabilityVSAvoidnucleic acid extractability
Core Design Contradiction:
Stability of the object's compositionVSLoss of substance

Solution Approach 1:

The patent extracts the harmful crosslinking function from the stabilization process by replacing formalin with liquid nitrogen freezing, thereby removing the cause of nucleic acid binding while maintaining tissue structure through physical preservation

Inventive Principle:
Principle #2Taking out (Extraction)

3Measurement precision

If cationic detergents are used to stabilize nucleic acids, then qualitative detection is improved, but the method is limited to samples with individual cells or single cell layer

Engineering Contradiction:
Improvenucleic acid detection precisionVSAvoidsample type adaptability
Core Design Contradiction:
Measurement precisionVSAdaptability or versatility

Solution Approach 1:

The patent creates a universal stabilization method using liquid nitrogen freezing that applies to all sample types (tissues, cells, body fluids) regardless of complexity, making the technique universally applicable while maintaining high nucleic acid preservation quality

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

4Stability of the object's composition

If highly concentrated ammonium sulfate is used to stabilize nucleic acids, then stabilization is achieved, but the method is unsuitable for body fluids and some tissue types

Engineering Contradiction:
Improvenucleic acid stabilizationVSAvoidsample type compatibility
Core Design Contradiction:
Stability of the object's compositionVSAdaptability or versatility

Solution Approach 1:

The patent changes the fundamental parameter of sample preservation from chemical stabilization (ammonium sulfate) to physical freezing at -70°C, which universally applies to all sample types including body fluids and diverse tissues without limitation

Inventive Principle:
Principle #35Parameter changes

5Stability of the object's composition

If freezing in liquid nitrogen is used, then nucleic acid and protein integrity is maintained, but complex logistical requirements and equipment costs are incurred

Engineering Contradiction:
Improvemolecular integrityVSAvoidstorage equipment complexity
Core Design Contradiction:
Stability of the object's compositionVSDevice complexity

Solution Approach 1:

The patent employs liquid nitrogen, a cheap and readily available cryogenic substance, as a disposable freezing medium that can be easily replaced, eliminating the need for expensive, complex refrigeration equipment while maintaining molecular integrity

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

6Duration of action of stationary object

If freezing is used to preserve samples, then long-term storage is enabled, but thawing during transport or processing leads to RNA degradation

Engineering Contradiction:
Improvestorage durationVSAvoidRNA integrity during processing
Core Design Contradiction:
Duration of action of stationary objectVSReliability

Solution Approach 1:

The patent performs preliminary stabilization by freezing samples in liquid nitrogen at -70°C immediately at the time of collection, creating a stable state that prevents subsequent degradation during transport and processing, eliminating the need for thawing

Inventive Principle:
Principle #10Preliminary action

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 enables the stabilization of biological samples at room temperature or refrigeration conditions, maintaining the integrity of nucleic acids and proteins, facilitating both qualitative and quantitative analysis, and allowing for storage without the use of liquid nitrogen or freezers, thus overcoming previous limitations in sample preservation and analysis.

Implementation Method 1

A method involving the use of polyol compositions, specifically polyethylene glycols and polypropylene glycols, to stabilize biological samples

Methodology Applied
Scientific EffectSolvation: Solvation

Data Source

PatentEP1969341B1A method for treating a biological sample
Publication Date: 2019.05.29 QIAGEN GMBH
  • EP1969341B1 patent drawingFigure 1~1b
  • EP1969341B1 patent drawingFigure 2
  • EP1969341B1 patent drawingFigure 3

AI summary

The invention relates to a method for treating a biological sample, comprising the following method steps: i) preparation of a biological sample and ii) bringing the biological sample into contact with a composition, comprising: (1) 1 to 100 wt. % of at least one polyol and (2) 0 to 99 wt. % of at least one additive, wherein the total amount of components (1) and (2) is 100 wt. %. The invention further relates to biological samples obtained by said method, a method for analysis of a treated biological sample, devices for treating a biological sample, use of said devices, various kits and use of a composition.