Nucleating-Lock Containers for Below-Freezing Degradation Kinetics
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Solution Overview
Problem
Existing isochoric methods for measuring protein degradation kinetics below freezing temperature face challenges such as complexity, lack of reproducibility, and incompatibility with industrial requirements, particularly in small-volume samples, due to issues with ice seed insertion and uncontrolled nucleation.
Innovation Solution
A method and apparatus using high-pressure containers with nucleating-locks that induce nucleation by closing below freezing temperature, eliminating the need for ice seeds, and a temperature-control holder for consistent temperature management, enabling high-throughput analysis with improved reproducibility.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If ice seeds are inserted to ensure controlled nucleation, then nucleation control is improved, but device complexity and ease of operation deteriorate due to the need for manual ice seed insertion and visual inspection
Solution Approach 1:
The container performs self-nucleation through its own structure: the removable bottom acts as a nucleation site when pulled out, eliminating the need for external ice seeds. The system serves itself by using its structural component (bottom) to trigger the required phase change (freezing nucleation).
Solution Approach 2:
The bottom of the container is extracted/removed to trigger nucleation. By pulling out the bottom, the system creates the nucleation condition without requiring separate ice seeds or external nucleation agents, simplifying the overall process.
2Reliability
If large aspect ratio high-pressure containers are used, then nucleation control is improved, but weight and ease of operation worsen due to heavy construction and difficulty in closing/opening
Solution Approach 1:
The container is segmented into a main body and a removable bottom portion. This segmentation allows the bottom to be pulled out for nucleation while keeping the main container structure compact and lightweight, avoiding the need for large aspect ratio containers.
Solution Approach 2:
The bottom of the container is made dynamic/removable rather than fixed. This allows the container to transition between closed (for pressurization) and open (for nucleation) states, providing nucleation control without requiring heavy permanent structural modifications.
3Measurement precision
If visual inspection and user experience are required to ensure no air is left inside, then measurement precision may be maintained, but productivity and ease of operation deteriorate due to time-consuming inspection
Solution Approach 1:
The filling process is self-regulating: air bubbles naturally rise and escape during filling, and the removable bottom design allows easy verification and removal of any remaining bubbles without requiring expert visual inspection.
Solution Approach 2:
Air bubbles are addressed preliminarily during the filling process itself. The filling method and removable bottom design work together to prevent bubble entrapment before the experiment begins, eliminating the need for post-filling inspection.
4Stability of the object's composition
If the container is filled to completely full liquid, then isochoric conditions are improved, but pressure control worsens due to abrupt pressure rise from uncontrolled nucleation
Solution Approach 1:
Nucleation is triggered preliminarily and controllably by removing the bottom before pressurization. This ensures that ice formation begins in a controlled manner, preventing abrupt pressure rises that would occur with uncontrolled nucleation after filling.
Solution Approach 2:
The removable bottom acts as an intermediary mechanism between the liquid and the freezing process. It provides a controlled interface for nucleation, allowing the system to transition from liquid to solid phase gradually and predictably, maintaining pressure stability.
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
Facilitates high-reproducibility, easy operation, and cleaning of protein degradation kinetics measurements in small volumes, addressing the limitations of existing isochoric methods by ensuring consistent thermal history and reducing laboriousness.
Implementation Method 1
closing the container with a nucleating-lock, wherein the nucleating-lock is at a temperature below the freezing temperature of the biomaterial and induces the nucleation
Implementation Method 2
an ice layer is formed acting as a natural piston and ensuring the isochoric conditions
Implementation Method 3
under isochoric conditions in high-pressure containers to prevent the formation of ice
Implementation Method 4
the low-density ice attempts to grow, but therefore compresses the solution, raising the pressure and suppressing the freezing line
Implementation Method 5
a temperature-control holder for consistent temperature management, enabling high-throughput analysis with improved reproducibility
Data Source
AI summary
This present disclosure describes an apparatus and a method for measuring degradation kinetics of a biomaterial in aqueous solution below the normal freezing temperature. Further includes new systems comprising high-pressure containers with nucleating-locks, a temperature-control holder for the high-pressure containers, a holder for the nucleating-locks and an insulated low-temperature gradient chamber. A method is described comprising several steps such as filling of the high-pressure container with the biomaterial solution to be slightly below freezing temperature, closing of the container with a nucleating-lock, which is below freezing temperature, allowing the container to reach the reaction temperature, removing the container at selected time and bringing it to above freezing temperature and opening the container to recover the biomaterial solution.