Solution Phase Isoelectric Focusing Device for Protein Prefractionation
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Solution Overview
Problem
Current analytical techniques, such as 2D PAGE, struggle to resolve the complexity of eukaryotic proteomes due to limited spatial resolution, leading to obscured detection of low abundance protein species, and existing devices for solution phase isoelectric focusing lack convenience and effectiveness for small volume protein samples.
Innovation Solution
A solution phase isoelectric focusing device with a loading tube assembly that uses screw cap compression for leak-proof seals, fixed pH membrane disks for proper orientation, and optional agitation to prevent air bubbles, allowing for efficient prefractionation of small volume protein samples.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional 2D PAGE is used to analyze complex protein mixtures, then the technique can handle protein samples, but the spatial resolution is limited causing high abundance species to obscure low abundance species
Solution Approach 1:
The device divides the protein sample into multiple separate chambers (first sample chamber, second sample chamber, third sample chamber) that are physically isolated from each other. Each chamber can be independently filled with buffer and protein sample, allowing proteins to be separated and analyzed in discrete, non-overlapping zones rather than competing for the same detection space in conventional 2D PAGE.
Solution Approach 2:
The patent introduces buffer chambers as intermediary elements between the sample chambers. These buffer chambers contain ampholyte buffers that create pH gradients to separate proteins based on their isoelectric points, acting as a mediating mechanism that enables resolution of protein species before they enter the detection phase, thereby preventing obscuration by high abundance species.
2Measurement precision
If existing solution phase IEF devices are used, then some prefractionation capability is achieved, but the devices are complex and inconvenient for small volume protein samples
Solution Approach 1:
The device is segmented into modular components: separate sample chambers, buffer chambers, and a loading tube assembly that can be independently assembled and disassembled. This modular segmentation reduces complexity by allowing users to configure only the necessary chambers for their specific sample volumes and protein types, making the device more convenient for small volume samples while maintaining prefractionation capability.
Solution Approach 2:
The device incorporates a movable loading tube assembly that can be inserted into and removed from the electrophoresis chamber, and the chambers can be independently filled and drained. This dynamic configuration allows the device to adapt to different sample volumes and experimental requirements, reducing the perceived complexity by enabling flexible, on-demand setup rather than requiring a fixed, complex instrument.
3Productivity
If protein samples are loaded into chambers, then analysis can proceed, but air bubbles may be trapped causing interference
Solution Approach 1:
The device includes a preliminary action step where the loading tube assembly is filled with buffer solution before the actual sample loading. This preliminary filling creates a bubble-free environment and establishes proper fluid pressure conditions, preventing air bubbles from being trapped during subsequent sample introduction and ensuring smooth, bubble-free protein sample loading.
Solution Approach 2:
The buffer chambers act as intermediary fluid pathways that mediate between the sample chambers and the electrophoresis chamber. By filling these intermediate buffer chambers first, the system creates a continuous fluid pathway that prevents air bubble entrapment during sample loading, as the buffer acts as a mediator that displaces air and establishes proper fluid dynamics.
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 effective prefractionation of complex protein samples, improving the resolution and detection of low abundance proteins by minimizing leaks and air bubbles, facilitating subsequent analytical techniques like 2D PAGE.
Implementation Method 1
The screw cap is tightened to ensure leak-proof seals between the chambers and other component parts
Implementation Method 2
Sealing O-rings between the chambers compress under the pressure to form leak-proof seals
Implementation Method 3
the device uses fixed pH membrane disks
Implementation Method 4
solution phase isoelectric focusing (IEF)
Implementation Method 5
the device is positioned on a rocker platform or a rotary platform to move the entire device during operation, thus effecting sample agitation
Implementation Method 6
move the entire device during operation, thus effecting sample agitation
Implementation Method 7
air bubbles are avoided in sample chambers by using optional specially shaped openings and cap seals to seal the chambers
Data Source
AI summary
A device for fluid phase electrophoresis, particularly solution phase isoelectric focusing, components thereof, and methods for use are presented.