Dialysis Membrane Substance Separation for Immunoassay
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Solution Overview
Problem
Current immunoassay methods require complex B/F separation processes, including antibody immobilization and washing, which are time-consuming and not suitable for simple analyses, especially for large molecular weight substances like proteins, and dialysis-based methods are limited by molecular size restrictions.
Innovation Solution
A substance separating device with a reaction vessel partitioned by a dialysis membrane, using dye-modified molecules that bind to the target substance, allowing for separation and detection without the need for immobilization, by exploiting the molecular weight cut-off of the dialysis membrane to retain the substance in one chamber while the dye-modified molecules can pass through, enabling quantitative measurement of high molecular weight substances.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If solid-phase method with antibody immobilization is used, then B/F separation can be achieved, but the process becomes complex and time-consuming requiring multiple steps including immobilization and washing
Solution Approach 1:
The invention extracts the essential separation function from the complex solid-phase process. By using a dialysis membrane with specific molecular weight cut-off, the method separates free dye-modified molecules from bound complexes without requiring antibody immobilization on solid phase, eliminating multiple washing and immobilization steps while maintaining separation capability
Solution Approach 2:
The invention changes the separation mechanism from solid-phase adsorption to size-based dialysis separation. By selecting a dialysis membrane with appropriate molecular weight cut-off (greater than dye-modified molecule weight but less than antigen-antibody complex weight), the method achieves separation based on molecular size parameters rather than surface immobilization
2Measurement precision
If solid-phase method with antibody immobilization is used, then B/F separation can be achieved, but time is required for immobilization and multiple processing steps
Solution Approach 1:
The invention removes the time-consuming antibody immobilization step entirely by using dialysis membrane separation. The method directly separates bound and free components based on molecular size through the dialysis membrane, eliminating the sequential time requirements for immobilization, washing, and separation that characterize solid-phase methods
Solution Approach 2:
The invention performs preliminary separation setup by pre-selecting a dialysis membrane with appropriate molecular weight cut-off characteristics. This preliminary configuration allows direct separation upon adding the dye-modified molecules to the specimen, without requiring preliminary immobilization steps
3Ease of operation
If dialysis-based B/F separation method is used, then washing is not required, but the method is not suited for measuring large molecular weight substances like proteins
Solution Approach 1:
The invention changes the dialysis membrane selection parameters by choosing a membrane with molecular weight cut-off specifically greater than the dye-modified molecule weight but less than the antigen-antibody complex weight. This parameter optimization enables the dialysis method to retain large molecular weight antigen-antibody complexes while allowing free dye-modified molecules to pass through, solving the limitation of conventional dialysis for large molecule measurement
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 allows for accurate and efficient separation and quantification of high molecular weight substances like proteins in immunoassays without the need for antibody immobilization, simplifying the analysis process and overcoming size limitations of dialysis-based methods.
Implementation Method 1
a reaction vessel provided with an interior that is partitioned into a first chamber and a second chamber by a dialysis membrane
Implementation Method 2
a dye-modified molecule that has a binding ability for specifically binding to a substance to be measured
Implementation Method 3
a detection optical system that emits, on at least one of the first chamber and the second chamber of the reaction vessel of the substance separating device, an excitation light that excites a dye and, also, detects an emission intensity, or the emission intensity and a degree of polarization
Data Source
AI summary
A substance separating device includes a reaction vessel provided with an interior that is partitioned into a first chamber and a second chamber by a dialysis membrane, a solution being stored in the reaction vessel. A specimen is stored in the first chamber, and dye-modified molecule that has a binding ability for specifically binding to a substance to be measured having a molecular weight greater than a molecular weight cut-off the dialysis membrane is stored in at least one of the first chamber and the second chamber. The dye-modified molecule has a molecular weight that is less than the molecular weight cut-off of the dialysis membrane.