Prion Protein Aggregation via Controlled Shear-Force

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Solution Overview

Problem

Current methods for producing aggregated conformation prion protein from native conformation prion protein are not reproducible due to dependence on ultrasound agitation, which varies with reaction vessel position and volume, leading to inconsistent yields of protease-resistant prion protein.

Innovation Solution

A device and method utilizing controlled shear-force application with a narrow intensity range (up to 10%) to fragment prion protein aggregates, ensuring consistent and reproducible generation of aggregated conformation prion protein, achieved by precise positioning of the liquid preparation relative to the shear-force generator and controlled shear-force intensity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If ultrasound agitation is used to produce aggregated conformation prion protein, then the conversion from native to aggregated conformation occurs, but the process becomes non-reproducible due to variation with reaction vessel position and volume

Engineering Contradiction:

Improvereproducibility of aggregated conformation productionVSAvoiddependence on reaction vessel position and volume

Core Design Contradiction:

ReliabilityVSDevice complexity

Solution Approach 1:

The patent replaces the ultrasound agitation system with a shear-force generator that produces controlled shear-force fields. This substitution eliminates the problems associated with ultrasound (position and volume dependence) while achieving the same goal of converting native conformation prion protein to aggregated conformation through a more controllable mechanical means.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the physical parameter used for agitation from ultrasound frequency and intensity to shear-force intensity with a narrow range (up to 10%). This parameter change enables reproducible production of aggregated conformation prion protein because shear-force can be precisely controlled and is less sensitive to reaction vessel variations.

Inventive Principle:

Principle #35Parameter changes

2Manufacturing precision

If controlled shear-force with narrow intensity range is applied, then reproducible aggregated conformation production is achieved, but the device complexity increases due to precise positioning requirements

Engineering Contradiction:

Improveuniformity of aggregated conformationVSAvoidprecise positioning of liquid preparation

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

The patent incorporates feedback mechanisms to monitor and adjust the position of the liquid preparation relative to the shear-force generator. This feedback system ensures that the liquid preparation remains in the optimal position for receiving controlled shear-force, thereby maintaining manufacturing precision without requiring manual intervention or complex mechanical positioning systems.

Inventive Principle:

Principle #23Feedback

3Productivity

If ultrasound agitation is used, then aggregation occurs, but the yield of protease-resistant prion protein varies inconsistently

Engineering Contradiction:

Improveyield of protease-resistant prion proteinVSAvoidconsistency of aggregation yield

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent changes the agitation parameter from ultrasound (which has wide intensity variations) to shear-force with a narrowly controlled intensity range (up to 10%). This parameter change directly addresses the inconsistency in aggregation yield by providing a more stable and controllable physical stimulus that produces consistent conversion rates from native to aggregated conformation.

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

The method produces aggregated conformation prion protein with uniform conformation and high reproducibility, as demonstrated by proteinase resistance and structure-sensitive spectroscopy, such as NMR, with specific shear-force intensities resulting in distinct conformation species and high conversion rates from native to aggregated conformation.

Implementation Method 1

subjecting a liquid preparation of native conformation prion protein in contact with aggregated conformation prion protein to at least one cycle or to a number of cycles of application of shear-force for fragmenting aggregates of prion protein

Methodology Applied

Scientific EffectShear-force: Shear Stress

Data Source

PatentUS10449499B2Device and method for production and analysis of prions

Publication Date: 2019.10.22 ALOIS DATA GMBH

AI summary

The invention provides a method for producing prion protein having an aggregated conformation by contacting native conformation prion protein with aggregated conformation prion protein in a liquid preparation and subjecting this to at least one cycle or to a number of cycles of application of shear-force for fragmenting aggregates of prion protein, wherein the shear-force applied is precisely controlled. In addition to this process for amplification of aggregated state prion protein from native conformation prion protein, the invention relates to the aggregated state prion protein obtained by the amplification process, which aggregated state prion protein has one conformation, which is e.g. identical within one batch and reproducible between batches, e.g. as detectable by proteinase resistance in a Western blot.