Biosensor Protein Stabilization Using Canavalia Lysate

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Solution Overview

Problem

Purified proteins used in biochemical sensors often have lower stability than in their natural environment, making it difficult to maintain their functionality due to disruptive impurities and environmental factors, which hinders the measurement of analyte-receptor interactions.

Innovation Solution

A cell lysate fraction from Canavalia ensiformis, enriched with precanavalin, is used to stabilize isolated proteins like concanavalin A, which is then incorporated into hydrogel particles with dextran to enhance protein stability and maintain receptor function.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Object-generated harmful factors

If proteins are purified from natural cells and tissues, then disruptive impurities are removed and targeted modification becomes possible, but the stability of the isolated protein decreases

Engineering Contradiction:
Improvedisruptive impuritiesVSAvoidprotein stability
Core Design Contradiction:
Object-generated harmful factorsVSStability of the object's composition

Solution Approach 1:

The patent extracts and removes disruptive impurities from the natural cellular environment through purification processes, separating the protein from harmful components while intentionally retaining or adding stabilizing components to maintain protein stability in the purified state

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent introduces intermediary substances (stabilizers such as sugars, polyols, proteins, or cell lysate fractions) that mediate between the purified protein and the surrounding environment, protecting the protein from destabilizing factors while allowing the protein to remain purified and modifiable

Inventive Principle:
Principle #24Intermediary (Mediator)

2Stability of the object's composition

If traditional stabilizers like bovine serum albumin are used, then protein stability is improved, but the stabilizer cannot withstand autoclaving for sterility assurance

Engineering Contradiction:
Improveprotein stabilityVSAvoidsterility assurance
Core Design Contradiction:
Stability of the object's compositionVSReliability

Solution Approach 1:

The patent changes the physical and chemical parameters of the stabilizer by selecting substances that can withstand autoclaving conditions (high temperature and pressure), such as certain sugars, polyols, and cell lysate fractions, which maintain their stabilizing function even after sterilization processes

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent employs stabilizers that are robust enough to survive autoclaving, treating the stabilizer as a component that can endure harsh sterilization conditions, thereby ensuring both sterility and continued functionality in the final product

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 cell lysate fraction significantly improves the stability of isolated proteins, allowing for effective use in biochemical sensors, particularly in glucose concentration determination, with precanavalin demonstrating better stabilization than traditional stabilizers like bovine serum albumin and enabling autoclaving for sterility.

Implementation Method 1

Low or high molecular weight additives are often used to reduce the free volume in the solution ('molecular crowding').

Methodology Applied
Scientific EffectMolecular crowding:

Implementation Method 2

The effect of these additions can be explained by the 'excluded volume' effect (Chebotareva et al., 2004, Biochemical Effects of Molecular Crowding, Biochemistry (Moscow), 69: 1522-1536).

Methodology Applied
Scientific EffectExcluded volume effect:

Implementation Method 3

The biochemical sensor particularly preferably additionally comprises dextran and concanavalin in hydrogel particles.

Methodology Applied
Scientific EffectHydrogel encapsulation: Hydrogel

Data Source

PatentEP2493915B1Stabilisation of biosensors for in vivo applications
Publication Date: 2017.08.16 EYESENSE AG

AI summary

The invention relates to the use of preparations for stabilizing isolated proteins. The invention in particular relates to the use of said preparations for stabilizing receptors in biochemical sensors. The invention further relates to biochemical sensors having said preparations.