Microfluidic Milk Classification Using Real-Time BRET Sensing
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Solution Overview
Problem
Existing methods for detecting analytes in samples, such as milk or cheese, face challenges with high costs, limited sensitivity, slow reaction times, and sensor drift, making real-time monitoring difficult and requiring surface regeneration.
Innovation Solution
A microfluidic method using a sensor molecule with a chemiluminescent donor and acceptor domain within a specific Förster distance, allowing real-time detection of analytes through binding analytes and substrates, and electro-optical sensing to modify the BRET ratio, enhancing sensitivity and reducing sensor drift.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If surface-based sensing techniques (SPR) are used, then real-time monitoring is enabled, but surface regeneration is required causing downtime
Solution Approach 1:
The patent extracts the sensing function from a surface-based system and transfers it to freely diffusing sensor molecules in solution. The sensor molecules are not attached to any surface but instead float freely in the microfluidic channel, allowing continuous monitoring without regeneration downtime. This is achieved by using fluorescently labeled sensor molecules that bind to analytes in solution phase.
Solution Approach 2:
The patent replaces the mechanical surface regeneration process with an optical detection system. Instead of physically regenerating a sensor surface, the system uses fluorescence resonance energy transfer (FRET) between freely diffusing sensor molecules and analytes, detected optically without mechanical intervention. This substitution eliminates the need for surface regeneration and associated downtime.
2Measurement precision
If conventional detection equipment is used, then BRET can be detected, but the equipment is sophisticated and costly
Solution Approach 1:
The patent creates a simplified optical copy of the BRET phenomenon by using fluorescently labeled sensor molecules where the fluorescent label acts as the acceptor. This allows detection using standard fluorescence microscopy equipment rather than specialized BRET detection systems. The fluorescent label copies the energy transfer function in a more accessible format.
Solution Approach 2:
The patent changes the detection parameter from measuring bioluminescence resonance energy transfer directly to measuring fluorescence intensity changes. By converting the BRET signal into a fluorescence signal that can be detected by standard equipment, the system maintains measurement precision while reducing device complexity and cost.
3Productivity
If microfluidic technologies are used, then reagent consumption is reduced and reaction rate is fast, but integration of multiple components increases cost per chip
Solution Approach 1:
The patent designs the microfluidic chip to perform multiple functions using a single integrated structure. The chip includes reservoirs, mixing chambers, and detection zones all in one device, eliminating the need for separate equipment for sample preparation, mixing, and detection. This multi-functionality reduces the overall system cost while maintaining fast reaction rates.
Solution Approach 2:
The patent merges multiple discrete components (sensor molecules, substrates, mixing mechanisms, and detection systems) into a single integrated microfluidic chip. The sensor molecules and substrates are combined in the same chamber where mixing and detection occur simultaneously, reducing the number of separate components and lowering per-chip costs.
4Quantity of substance
If electronic noses and tongues are used, then analyte detection is performed, but sensor selectivity and sensitivity are poor
Solution Approach 1:
The patent uses different fluorescently labeled sensor molecules with specific binding properties for different analytes. Each sensor molecule has localized specificity for particular targets (e.g., lactose, proteins, bacteria), allowing highly selective detection within the complex milk matrix. This local quality of sensor specificity overcomes the poor selectivity of electronic nose sensors.
Solution Approach 2:
The patent employs a composite sensing system combining multiple fluorescently labeled sensor molecules, each with different binding specificities. This composite approach allows simultaneous detection of multiple analytes with high selectivity and sensitivity, overcoming the limitations of single-type sensors in electronic noses and tongues.
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 method achieves rapid, sensitive, and real-time detection of analytes with improved signal-to-noise ratio, reducing downtime and costs, and enabling precise concentration coding.
Implementation Method 1
a chemiluminescent donor domain and an acceptor domain
Implementation Method 2
RET is a ratiometric technique which can eliminate data variability caused by fluctuations in light output due to variations in assay volume, assay conditions and signal decay across different wells in a plate
Implementation Method 3
a domain that binds one or more analytes
Data Source
Figure 1
Figure 2A~2B
Figure 3~4(c)
AI summary
The present invention relates to methods and systems for detecting one or more analytes in a sample and/or for classifying a sample. In particular, the present invention relates to methods and systems which can be used to detect the analytes in real time and which rely on flowing through a microfluidic device one or more types of sensor molecule each comprising a domain that binds one or more analytes, a chemiluminescent donor domain and an acceptor domain, wherein the separation and relative orientation of the chemiluminescent donor domain and the acceptor domain, in the presence and/or absence of analyte is within ± 50% of the Forster distance.