Multi-compartment Filter for Rapid Protein pI Separation
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Solution Overview
Problem
Conventional methods for separating proteins based on their isoelectric points are slow and inefficient, requiring hours to complete, which is unacceptable for preparative purposes.
Innovation Solution
A filter apparatus with a central reservoir and satellite reservoirs, each equipped with filtration devices of varying pI-selectivity, allows for the rapid separation of proteins by applying an electric field and using centrifugal force to move molecules into different reservoirs based on their pI values, with zwitterionic barriers and isoelectric bead beds facilitating the separation process.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional gel electrophoresis or isoelectric focusing is used to separate proteins, then separation accuracy is maintained, but separation time becomes excessively long (hours)
Solution Approach 1:
The device divides the separation process into multiple compartments (first compartment with anode, second compartment with cathode, and additional compartments in between) separated by membranes. Each compartment contains filtration devices with different pI-selectivity values, allowing simultaneous separation of multiple protein fractions through the membranes, thereby reducing overall separation time while maintaining accuracy
Solution Approach 2:
The invention transitions from one-dimensional linear migration in conventional gels to multi-dimensional separation by adding vertical stacking of compartments with different pI-selectivity membranes. This allows proteins to be separated simultaneously along multiple selective dimensions (different pI ranges) rather than sequentially, dramatically reducing separation time
2Productivity
If multi-compartment electrolyzers with isoelectric membranes are used for preparative protein purification, then processing capacity increases, but system complexity increases
Solution Approach 1:
The device is divided into modular compartments that can be independently assembled and configured. Each compartment contains specific filtration devices with defined pI-selectivity ranges, allowing the system to be scaled by adding or removing compartments based on processing requirements, thus managing complexity through modularity
Solution Approach 2:
The system uses filtration devices with varying pI-selectivity parameters (different isoelectric point ranges) to achieve separation. By changing the pI-selectivity parameter of the membranes rather than redesigning the entire system, the device can handle different protein types and processing capacities
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
This method enables quick and accurate separation, concentration, and fractionation of proteins, reducing the separation time significantly and improving the efficiency of protein purification.
Implementation Method 1
When the electrodes are charged, each of the protein molecules travels toward one of the electrodes, according to their net charge at the pH of the buffered polyacrylamide gel
Implementation Method 2
Isoelectric focusing is an electrophoresis technique in which proteins move under an electric field through a pH gradient. All proteins migrate towards the cathode or the anode until they encounter a pH identical to their isoelectric point. At this point the protein loses its charge and stops moving
Implementation Method 3
a plurality of filtration devices, the devices differing from each other in pI-selectivity, wherein each device is positioned in a portal
Data Source
AI summary
Each embodiment includes a central sample reservoir and a plurality of satellite reservoirs. In a first embodiment, a first electrode in electrical contact with the central reservoir is charged and second electrodes in electrical contact with the satellite reservoirs are sequentially charged, thereby pI filtering molecules in the central reservoir into the satellite reservoirs. In a second embodiment, the central reservoir is configured to rotate so that molecules in a sample in the central reservoir are centrifugally pI-filtered into the satellite reservoirs. In a third embodiment, first and second electrodes proximate opposite first and second satellite reservoirs, respectively, are charged. Some molecules in a sample are pI filtered into the first and second satellite reservoirs. Third and fourth electrodes proximate opposite third and fourth satellite reservoirs, respectively, are then charged. Some molecules in a sample are pI filtered into the third and fourth satellite reservoirs.