Magnetic Microparticle Biomolecule Analysis Method
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Solution Overview
Problem
Conventional biomolecule analysis methods, such as DNA microarrays and ELISA, have limitations including a narrow dynamic range, slow reaction times, and inability to accurately measure absolute biomolecule concentrations, which can lead to overlooking crucial biomolecules expressed at trace levels.
Innovation Solution
A method involving immobilization of biomolecules on magnetic microparticles, allowing for rapid reaction with probe molecules in a dispersed state, followed by fluorescence measurement, enabling single-molecule resolution and comprehensive analysis of thousands of biomolecule types with a dynamic range of 4 digits or more without amplification reactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If probe molecules are immobilized on support substrate to perform specific binding reaction, then specific detection of biomolecules is achieved, but reaction time becomes very long (10 hours or more)
Solution Approach 1:
The patent inverts the conventional approach by immobilizing biomolecules on magnetic microparticles instead of immobilizing probe molecules on support substrate. This inversion allows the sample to be added directly to the magnetic microparticles, enabling rapid binding without long incubation periods while maintaining specific detection capability.
Solution Approach 2:
The patent utilizes magnetic field (analogous to pneumatic/hydraulic principles) to control and manipulate magnetic microparticles. By applying magnetic fields, the microparticles can be rapidly concentrated, separated, and immobilized on support substrate, dramatically reducing reaction time compared to conventional diffusion-based methods.
2Adaptability or versatility
If multiple probe molecules are immobilized on support substrate, then comprehensive analysis is achieved, but dynamic range is limited to 2-3 digits
Solution Approach 1:
The patent segments the analysis by using individual magnetic microparticles, each carrying a single biomolecule, rather than having multiple probe molecules on a flat substrate. This segmentation enables single-molecule resolution and extends the dynamic range to 4 digits or more by allowing accurate counting of individual biomolecule binding events.
Solution Approach 2:
The patent transitions from two-dimensional immobilization on support substrate to three-dimensional suspended magnetic microparticles. This dimensional change allows for better spatial distribution, reduced background noise, and improved signal-to-noise ratio, thereby expanding the measurable dynamic range.
3Measurement precision
If probe molecules are immobilized on support substrate, then specific binding reaction is achieved, but reaction site is crowded causing slow reaction kinetics
Solution Approach 1:
The patent inverts the conventional configuration by placing biomolecules on magnetic microparticles instead of probe molecules on support substrate. This creates a suspended, three-dimensional reaction environment that eliminates crowding effects and enhances reaction kinetics while maintaining specific binding capability.
4Measurement precision
If conventional fluorescence measurement is used, then relative comparison between samples is achieved, but absolute number of biomolecules cannot be measured
Solution Approach 1:
The patent replaces conventional fluorescence intensity measurement with a counting-based detection method using magnetic microparticles. By detecting and counting individual microparticles that have bound biomolecules, the system directly quantifies the absolute number of biomolecules rather than relying on relative fluorescence intensity comparisons.
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 approach allows for rapid and sensitive analysis of biomolecules with high comprehensiveness and quantitativeness, providing a dynamic range that matches actual biomolecule abundance and enabling accurate detection of trace biomolecules.
Implementation Method 1
the microparticles are collected and immobilized by using magnetic force on a support substrate
Implementation Method 2
so as to be detected and evaluated by fluorescence measurement or the like
Data Source
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AI summary
An object of the present invention is to provide a method and means for analyzing biomolecules that can realize, in biomolecule analysis, a wide dynamic range attained by counting the number of biomolecules and rapid analysis. The present invention relates to a method for analyzing biomolecules, comprising the steps of: immobilizing biomolecules 101 to be analyzed on surfaces of magnetic microparticles 108; reacting labeled probe molecules 104 with the biomolecules 101 to be analyzed; collecting and immobilizing the microparticles 108 on a support substrate 110; and measuring a label on the support substrate 110. Since single-molecule immobilized magnetic microparticles are used in the present invention, the number of biomolecules can be counted, and since hybridization and an antigen-antibody reaction are performed with the microparticles having biomolecules immobilized thereon dispersed, the reaction can be rapidly performed. Further, the type and the abundance of biomolecules of interest can be determined at a single molecular level, so as to evaluate, in particular, the absolute concentration of biomolecules.