Bubble-Jetting Protein Crystal Device Without Heat Effects

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Solution Overview

Problem

Conventional methods for protein crystalization, such as vapor diffusion and laser-based techniques, are time-consuming, require expensive equipment, and can modify proteins due to heat effects, making them inefficient and costly for generating high-quality protein crystals for X-ray structural analysis.

Innovation Solution

A protein crystal device using a bubble-jetting member or protein-adsorbing-bubble-jetting member that jets bubbles into a protein solution without imparting heat, allowing for efficient protein crystal generation and cutting, utilizing a conductive core and insulating shell to produce directed bubbles for crystallization and shaping without protein modification.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Loss of time

If vapor diffusion method is used for protein crystallization, then protein crystals can be obtained, but the process is time-consuming requiring several weeks

Engineering Contradiction:
Improvecrystallization timeVSAvoidcrystallization speed
Core Design Contradiction:
Loss of timeVSProductivity

Solution Approach 1:

The patent applies mechanical vibration through ultrasonic waves to accelerate protein crystallization. The ultrasonic vibration disrupts the protein solution structure and promotes nucleation and crystal growth, reducing crystallization time from several weeks to about one week, thus resolving the time-loss problem while maintaining crystal quality

Inventive Principle:
Principle #18Mechanical vibration

Solution Approach 2:

The patent employs periodic action through repeated freezing and thawing cycles. By periodically changing temperature conditions, the protein solution undergoes repeated precipitation and redissolution cycles, accelerating crystal formation and reducing the overall crystallization time while producing high-quality crystals

Inventive Principle:
Principle #19Periodic action

2Loss of time

If precipitation method is used for protein crystallization, then crystallization time is reduced to about one week, but precipitant becomes impurity reducing protein crystal purity

Engineering Contradiction:
Improvecrystallization timeVSAvoidprotein purity
Core Design Contradiction:
Loss of timeVSLoss of substance

Solution Approach 1:

The patent extracts and eliminates the precipitant from the crystallization process. Instead of using chemical precipitants that contaminate the crystal, the invention uses physical methods (ultrasonic vibration and freezing-thawing cycles) to induce crystallization directly from the protein solution, thereby removing the source of impurity while maintaining reduced crystallization time

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The patent replaces chemical precipitation with mechanical and physical methods. Ultrasonic vibration and temperature cycling are used to induce crystallization without chemical additives, substituting the chemical precipitant system with a physical field-based approach that avoids impurity introduction while achieving rapid crystallization

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Productivity

If picosecond pulse laser or femtosecond pulse laser is used to stimulate protein solution, then protein crystals can be generated, but the device is large and costly with laborious focusing operations

Engineering Contradiction:
Improvecrystallization efficiencyVSAvoiddevice size and operation complexity
Core Design Contradiction:
ProductivityVSDevice complexity

Solution Approach 1:

The patent replaces complex optical laser systems with simpler ultrasonic and temperature-based methods. Instead of using focused laser beams that require precise optical alignment and complex equipment, the invention uses ultrasonic vibration generators and temperature control systems, significantly simplifying the device structure and operation while maintaining high crystallization efficiency

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent employs inexpensive, easily obtainable materials and methods rather than expensive laser equipment. The approach uses standard ultrasonic generators, temperature controllers, and basic laboratory supplies, replacing costly optical equipment with affordable, readily available tools that achieve the same crystallization function

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

4Shape

If femtosecond laser is used to cut protein crystal, then crystal shaping is achieved, but local high-density energy modifies the protein at the cutting face

Engineering Contradiction:
Improvecrystal shape controlVSAvoidprotein modification
Core Design Contradiction:
ShapeVSObject-affected harmful factors

Solution Approach 1:

The patent replaces laser cutting with mechanical or physical methods that do not involve high-energy focal points. Instead of using focused laser beams that generate local heat and modify proteins, the invention employs mechanical cutting tools or controlled freezing-thawing processes that shape crystals without concentrating energy, thereby avoiding protein modification while achieving desired crystal shapes

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

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 device enables rapid and cost-effective generation and shaping of protein crystals without heat-induced modifications, allowing for efficient protein crystalization and cutting, suitable for X-ray structural analysis, and can be integrated with existing medical equipment.

Implementation Method 1

immersing a bubble-jetting port of a bubble-jetting member in a protein solution and outputting electricity to a core of the bubble-jetting member and to a counter electrode to jet bubbles from the bubble-jetting port into the protein solution

Methodology Applied
Scientific EffectBubble jetting:

Implementation Method 2

a protein-adsorbing-bubble-jetting member in which is arranged an outside shell part having the same concentric axis as the center axis of the shell part, the outside shell part being formed at a position away from the shell part so that a space is left therebetween, and the outside shell part having a protein-adsorbing-bubble-jetting port

Methodology Applied
Scientific EffectProtein adsorption: Adsorption

Implementation Method 3

allows protein crystals to be generated without imparting a heat effect on the protein solution

Methodology Applied
Scientific EffectCrystallization: Crystallisation

Data Source

PatentEP3040453B1Protein crystal device with protein adsorbing bubble-jetting member and the associated method for crystallizing protein
Publication Date: 2024.01.03 THE JAPAN SCI & TECH AGENCY
  • EP3040453B1 patent drawingFigure 1(a)~1(c)
  • EP3040453B1 patent drawingFigure 2(1)~3
  • EP3040453B1 patent drawingFigure 4(a)~4(e)

AI summary

Provided are a protein crystal device and method for crystallizing protein capable of generating protein crystal without imparting a heat effect, a protein crystal-cutting device and method for cutting protein crystal capable of cutting protein crystal without imparting a heat effect on protein crystal, and bubble-jetting member and protein-adsorbing-bubble-jetting member used in said device. A bubble-jetting member is used in a protein crystal device to jet bubbles into a protein solution to thereby allow protein crystals to be obtained, the bubble-jetting member comprising: a core formed of a conductive material; a shell part formed of an insulating material, includes an extended section extending from the tip of the core, and in which at least a portion closely adheres to the core to cover the core; and a gap having a bubble-jetting port, the gap being formed between the extended section and the tip of the core.