Biomolecule Detection via Sample Fractionation and Microsphere Immobilization
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Solution Overview
Problem
Current methods for detecting proteins from small biological samples are limited by sensitivity, cost, and the need for complex sample preparation, particularly when trying to detect multiple proteins simultaneously, as they often suffer from cross-reactivity and require expensive mass spectrometry equipment.
Innovation Solution
A method involving the separation of biological samples by physical properties, followed by immobilization of biomolecules on distinguishable microsphere populations for multiplex immunoassays, allowing for sensitive and specific detection of hundreds of proteins from small sample volumes without the need for mass spectrometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If mass spectrometry is used for protein detection, then a large number of different proteins can be detected in an unambiguous manner, but the sensitivity is limited and the cost is very high
Solution Approach 1:
The biological sample is separated into multiple fractions based on physical properties such as size, charge, or hydrophobicity. Each fraction is then analyzed separately using immunological detection methods, allowing comprehensive protein detection while maintaining high sensitivity for each fraction.
Solution Approach 2:
The patent introduces fractionation as an intermediary step between sample preparation and detection. This intermediary process divides the complex sample into manageable fractions, enabling the use of highly sensitive but lower-throughput immunological methods to achieve both high sensitivity and comprehensive protein detection.
2Measurement precision
If mass spectrometry is used for specific protein detection, then unambiguous detection is achieved, but complicated and cost-intensive sample preparation and separation techniques are required
Solution Approach 1:
The patent employs relatively simple and inexpensive fractionation techniques (such as gel electrophoresis or chromatography) instead of complex mass spectrometry-based separation. These fractionation methods are easier to perform, require less specialized equipment, and can be completed in a single experimental workflow.
3Ease of operation
If immunoassays are used for protein detection, then specific detection without complicated sample preparation is achieved, but sensitivity is lowered when more than about 20 proteins are simultaneously detected
Solution Approach 1:
By dividing the complex biological sample into multiple fractions based on physical properties, the patent enables parallel immunological detection of many proteins. Each fraction contains a subset of proteins that can be detected with high sensitivity using standard immunoassay techniques, and the results are integrated to provide comprehensive protein detection.
4Productivity
If protein microarrays are used for multiplex detection, then dozens to hundreds of different proteins can be detected from small sample volumes, but cross-reactivity of antibodies limits sensitivity
Solution Approach 1:
The patent segments the protein detection problem by fractionating the sample beforehand. This reduces the complexity of each individual detection assay, allowing antibodies to be more specific in each fraction while still achieving comprehensive coverage across all fractions. The segmentation approach minimizes cross-reactivity issues by reducing the number of proteins present in each detection reaction.
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 enables the detection and quantification of numerous proteins with high sensitivity and specificity, reducing costs and complexity, while allowing for the analysis of post-translational modifications and multiplex analysis from limited sample amounts, such as tumor tissue.
Implementation Method 1
separating the biological sample into fractions according to at least one physical property of the biomolecules
Implementation Method 2
separating the biological sample into fractions according to at least one physical property of the biomolecules
Implementation Method 3
immobilizing the biomolecules from the individual fractions on microsphere populations specific for each fraction
Implementation Method 4
specifically detecting biomolecules present in the fractions using at least one solid-phase-based, immunological detection method
Data Source
AI summary
A method of detecting biomolecules present in a complex biological sample includes a) separating the biological sample into fractions according to at least one physical property of the biomolecules; and b) specifically detecting biomolecules present in the fractions using at least one solid-phase-based, immunological detection method including immobilizing the biomolecules from the individual fractions on microsphere populations specific for each fraction and distinguishable from one another.