Melezitose Substrate for Biomolecule Stabilization

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Solution Overview

Problem

Current dry-state technologies for preserving biomolecules like proteins and peptides are inefficient, requiring pre-purification and additional processing steps, and often result in denaturation and degradation, especially when stored at ambient temperatures, limiting their recovery and stability for downstream applications.

Innovation Solution

A solid substrate comprising melezitose or other trisaccharides, optionally with lysis and nucleic acid denaturing reagents, that allows for the direct collection, stabilization, and elution of biomolecules from biological samples in a dry state at ambient temperatures without pre-purification, using a single process step.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Temperature

If dry-state technologies are used for preserving proteins and peptides, then storage at ambient temperatures is enabled, but pre-purification and additional processing steps are required

Engineering Contradiction:
Improvestorage temperatureVSAvoidprocessing steps
Core Design Contradiction:
TemperatureVSDevice complexity

Solution Approach 1:

The patent combines collection, stabilization, and storage functions into a single substrate system. The substrate integrates drying capability with protein stabilization through incorporated excipients, eliminating the need for separate pre-purification and processing steps while enabling ambient temperature storage.

Inventive Principle:
Principle #5Merging (Combining)

Solution Approach 2:

The substrate is pre-prepared with stabilizing excipients and drying agents incorporated into its structure before use. This preliminary preparation allows the substrate to immediately stabilize proteins upon contact and provides the drying function needed for ambient temperature storage without requiring subsequent processing steps.

Inventive Principle:
Principle #10Preliminary action

2Stability of the object's composition

If proteins are stored in solution states, then structural and functional integrity is maintained, but refrigeration is required to prevent degradation

Engineering Contradiction:
Improveprotein integrityVSAvoidstorage temperature
Core Design Contradiction:
Stability of the object's compositionVSTemperature

Solution Approach 1:

The patent changes the physical state parameter from solution to dry state, and incorporates stabilizing excipients that modify the chemical environment. This parameter change allows proteins to maintain structural and functional integrity at higher temperatures by preventing denaturation and degradation through the stabilizing matrix.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The substrate acts as an intermediary between the protein and the ambient environment. The incorporated excipients and stabilizing agents mediate the interaction, protecting the protein from degradation while allowing ambient temperature storage without requiring refrigeration.

Inventive Principle:
Principle #24Intermediary (Mediator)

3Stability of the object's composition

If chemical additives are used to inhibit protease activity, then protein degradation is prevented, but down-stream analytical techniques are affected

Engineering Contradiction:
Improveprotein stabilityVSAvoidanalytical technique accuracy
Core Design Contradiction:
Stability of the object's compositionVSReliability

Solution Approach 1:

The patent changes the stabilization mechanism from chemical inhibition of proteases to physical stabilization through the substrate matrix and excipients. This parameter change prevents protein degradation through adsorption and structural stabilization rather than chemical protease inhibition, thereby avoiding interference with downstream analytical techniques.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The substrate serves as a disposable collection and stabilization device that can be discarded after use. This eliminates the need for complex chemical additives that would require removal before analysis, as the substrate itself provides the stabilization function without interfering with subsequent analytical measurements.

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

4Ease of manufacture

If untreated cellulose paper substrates are used for preservation, then simple storage is enabled, but analyte recovery and biological activity are insufficient

Engineering Contradiction:
Improvesubstrate simplicityVSAvoidanalyte recovery
Core Design Contradiction:
Ease of manufactureVSReliability

Solution Approach 1:

The patent creates a composite substrate by incorporating excipients and stabilizing agents into the cellulose paper matrix. This composite material maintains the simplicity and ease of manufacture of basic paper substrates while adding the functional properties needed for efficient analyte recovery and preservation of biological activity.

Inventive Principle:
Principle #40Composite materials

Solution Approach 2:

The substrate incorporates stabilizing properties at the local level where proteins contact the paper surface. The excipients are integrated into the substrate structure to provide localized stabilization and recovery enhancement exactly where needed, without complicating the overall substrate design or manufacture.

Inventive Principle:
Principle #3Local quality

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 substrate effectively stabilizes and retains the integrity and function of proteins and peptides, achieving higher recovery rates and prolonged storage without denaturation, as demonstrated by enhanced elution efficiency and stability compared to unmodified cellulose substrates.

Implementation Method 1

The substrate effectively stabilizes and retains the integrity and function of proteins and peptides, achieving higher recovery rates and prolonged storage without denaturation

Methodology Applied
Scientific EffectMolecular stabilization:

Implementation Method 2

A solid substrate comprising melezitose or other trisaccharides, optionally with lysis and nucleic acid denaturing reagents, that allows for the direct collection, stabilization, and elution of biomolecules from biological samples

Methodology Applied
Scientific EffectAdsorption: Adsorption

Implementation Method 3

eluting the biomolecules in a substantially intact form thereafter for further analysis

Methodology Applied
Scientific EffectSolubilization: Solvation

Data Source

PatentEP3134441B1Substrates and methods for collection, stabilization and elution of biomolecules
Publication Date: 2019.12.18 GLOBAL LIFE SCI SOLUTIONS OPERATIONS UK LTD
  • EP3134441B1 patent drawingFigure 1

AI summary

A solid substrate for the extraction, stabilization, and storage of proteins is provided. The substrate includes: a polysaccharide, such as melezitose under a substantially dry state. The substrate is configured to extract proteins from a sample and stabilize the extracted proteins in a dry format under ambient conditions for a prolonged period of time. Methods for collecting and recovering the proteins stored in the dry solid substrate are also described.