Poly(Proline) Ice Recrystallization Inhibition for Cryopreservation

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Solution Overview

Problem

Current cryopreservation methods using DMSO as a cryoprotectant are toxic and lead to phenotypic changes in cells, with low survival rates, and existing antifreeze proteins (AF(G)Ps) are toxic or immunogenic, while synthetic polymers like PVA have limited effectiveness and ice morphology issues.

Innovation Solution

Utilizing poly(proline) or its variants to inhibit ice recrystallization during cryopreservation, which can be used in compositions to treat biological materials and food products, forming a poly(proline) II type helix to effectively prevent ice crystal growth.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If DMSO is used as a cryoprotectant to prevent ice damage, then cell protection is improved, but cell toxicity and phenotypic changes increase

Engineering Contradiction:

Improvecell protectionVSAvoidcell toxicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent introduces poly(proline) as an intermediary substance that mediates between ice crystals and cells. Poly(proline) adsorbs to ice crystal surfaces and inhibits recrystallization, providing protection without the toxic effects of DMSO. This intermediary approach allows ice growth inhibition while maintaining cell viability and avoiding phenotypic changes.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent extracts the specific function of DMSO (ice growth inhibition) and separates it from its harmful toxic effects by using poly(proline) instead. This substitution removes the toxic component while retaining the protective function against ice recrystallization damage.

Inventive Principle:

Principle #2Taking out (Extraction)

2Reliability

If antifreeze proteins are used to inhibit ice recrystallization, then ice growth is prevented, but toxicity and immunogenicity increase

Engineering Contradiction:

Improveice recrystallization inhibitionVSAvoidtoxicity and immunogenicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent replaces complex, potentially immunogenic antifreeze proteins with a simpler, synthetic poly(proline) polymer. This synthetic polymer provides the necessary IRI function without the complexity and potential immunogenicity of natural proteins, making it safer for therapeutic applications.

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

Solution Approach 2:

The patent changes the chemical parameters from protein-based antifreeze agents to a synthetic poly(amino acid) structure. This parameter change from complex protein to simpler polymer maintains the IRI function while eliminating toxicity and immunogenicity concerns.

Inventive Principle:

Principle #35Parameter changes

3Reliability

If high concentrations of cryoprotectants are used to protect cells, then cell survival is improved, but toxicity increases

Engineering Contradiction:

Improvecell survivalVSAvoidcryoprotectant concentration

Core Design Contradiction:

ReliabilityVSQuantity of substance

Solution Approach 1:

The patent changes the chemical parameters of the cryoprotectant from DMSO to poly(proline), which has different toxicity characteristics. This allows for effective cryoprotection at concentrations that do not cause the same level of toxicity as traditional cryoprotectants.

Inventive Principle:

Principle #35Parameter changes

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

Poly(proline) significantly inhibits ice recrystallization at lower concentrations than previously thought, enhancing cell viability in biological materials and improving texture and flavor in frozen food products by minimizing ice crystal growth.

Implementation Method 1

poly(proline) has significant IRI activity and hence that it can be used for cryopreservation

Methodology Applied

Scientific EffectIce recrystallization inhibition:

Implementation Method 2

forming a poly(proline) II type helix to effectively prevent ice crystal growth

Methodology Applied

Scientific EffectHelix formation: Helix

Implementation Method 3

It is hypothesized that PVA may function by recognition between the regularly-spaced hydroxyl groups and the growing ice crystal

Methodology Applied

Scientific EffectMolecular recognition:

Implementation Method 4

Antifreeze (glyco)proteins (AF(G)Ps) from extremophile species are potent ice recrystallization inhibitors

Methodology Applied

Scientific EffectAdsorption: Adsorption

Data Source

PatentEP3634126B1Recrystallization inhibitor

Publication Date: 2025.08.27 UNIVERSITY OF WARWICK

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AI summary

The present invention relates to methods for preventing or inhibiting ice recrystallisation in substances (e.g. biological materials and food products) which are susceptible to ice crystal growth upon cryopreservation and/or thawing therefrom. The methods relate to the use of compositions comprising poly(proline) or a variant or derivative thereof. Also provided are kits and compositions comprising poly(proline) which can be used in the methods of the invention.