Modified CAHS Proteins for Gel-Free Biological Stabilization

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Solution Overview

Problem

Existing stabilizing agents for biological materials, such as intrinsically disordered proteins from tardigrades, tend to polymerize and form unwanted gel-like substances, limiting their effectiveness in extreme conditions.

Innovation Solution

Modified intrinsically disordered proteins, particularly tardigrade cytoplasmic abundant heat soluble (CAHS) proteins, are used to stabilize biological materials by preventing polymerization and forming gels, allowing storage under extreme conditions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If intrinsically disordered proteins (TDPs) are used as stabilizing agents, then stability of biological material is improved, but the proteins polymerize and form unwanted gel-like substances

Engineering Contradiction:

Improvestability of biological materialVSAvoidpolymerization and gel formation

Core Design Contradiction:

ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The patent applies segmentation by dividing the TDP sequence into multiple smaller peptide fragments (e.g., 5-20 amino acids each). These fragments are then combined in mixtures to achieve stabilization without the polymerization issues of full-length TDPs. The segmentation breaks up the long-range interactions that cause gel formation while retaining the stabilizing functionality in smaller units.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

The patent extracts specific functional domains or motifs from the full-length TDP sequences that are responsible for stabilization activity. By taking out only these essential regions and using them as truncated peptides, the invention achieves stabilization without the problematic regions that cause polymerization and gelation.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 3:

The patent changes key parameters of the TDP molecules by truncating them to shorter lengths (e.g., 10-50 amino acids) and adjusting their concentration ranges. These parameter changes modify the physical-chemical properties to eliminate gel formation while preserving the stabilizing effect on biological materials.

Inventive Principle:

Principle #35Parameter changes

2Ease of manufacture

If conventional stabilizing agents are used, then ease of manufacture is improved, but stability in extreme conditions deteriorates

Engineering Contradiction:

Improveease of manufactureVSAvoidstability in extreme conditions

Core Design Contradiction:

Ease of manufactureVSReliability

Solution Approach 1:

The patent uses inexpensive peptide fragments derived from TDPs that can be produced through simple recombinant expression or chemical synthesis. These short peptides are easier and cheaper to manufacture than full-length proteins, while still providing the necessary stabilization in extreme conditions through their disordered structure and ability to form protective complexes.

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 modified CAHS proteins effectively protect and preserve biological materials under non-ideal conditions, maintaining structural and functional integrity over a wide temperature range.

Implementation Method 1

intrinsically disordered proteins (IDPs) from extremophilic and extremotolerant organisms as stabilizing agents

Methodology Applied

Scientific EffectIntrinsic disorder:

Implementation Method 2

at concentrations utilized for stabilization, TDPs begin to polymerize and self-associate to form unwanted gel-like substances

Methodology Applied

Scientific EffectPolymerization prevention:

Implementation Method 3

maintaining structural and functional integrity over a wide temperature range

Methodology Applied

Scientific EffectThermal stabilization:

Implementation Method 4

the extreme stress tolerance mechanisms of living organisms such as tardigrades, plants, rotifers, and bacteria have been studied

Methodology Applied

Scientific EffectExtreme stress tolerance:

Data Source

PatentUS12410217B2Proteins for stabilization of biological material

Publication Date: 2025.09.09 UNIVERSITY OF WYOMING

AI summary

Embodiments of the present disclosure generally relate to methods and compositions for stabilizing biological material using intrinsically disordered proteins. In an embodiment, a composition is provided, the composition including a first component comprising at least one intrinsically disordered protein; and a second component comprising at least one biological material of interest, at least one biologically-derived material of interest, or both, the second component being free of the at least one intrinsically disordered protein. The methods and compositions include at least one intrinsically disordered protein that can be modified to prevent, or at least mitigate, polymerization thereof and the formation of gel-like matrices, thereby, e.g., improving the ability of the intrinsically disordered proteins to protect and stabilize sensitive biological materials.