Biosensor Dual Reaction Regions SPR Binding Activity
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Solution Overview
Problem
Conventional biosensors cannot simultaneously detect the binding of a compound to a functional protein and assay the reaction product derived from the protein's activity without using labels or fluorescence, limiting their applicability in drug screening and protein interaction studies.
Innovation Solution
A biosensor design featuring a first reaction region with a physiologically active substance immobilized for binding with test molecules and a second reaction region with molecules specific to the reaction products, allowing for simultaneous detection of binding and activity influences without labels or fluorescence, using a surface plasmon resonance biosensor with separated reaction regions and an assay region for dielectric constant changes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If only binding detection based on SPR signals is used, then the detection method is simple and label-free, but it cannot determine whether binding occurs at the active site or to inactivated proteins
Solution Approach 1:
The sensor chip surface is divided into two distinct reaction regions: a first reaction region for binding detection and a second reaction region for activity detection. This segmentation allows simultaneous acquisition of both binding information and functional activity information without requiring separate experiments or complex sample preparation.
Solution Approach 2:
The biosensor system performs multiple functions simultaneously using a single integrated platform: it detects both compound binding to proteins and the functional activity of those proteins through enzyme reactions. This multi-functionality eliminates the need for separate binding assays and activity assays, providing comprehensive information in one experiment.
2Measurement precision
If binding and reaction product assay are performed separately, then each measurement can be optimized, but the overall process becomes complex and time-consuming
Solution Approach 1:
The patent combines binding detection and reaction product assay into a single integrated biosensor system with two reaction regions on the same sensor chip. This merging allows simultaneous measurement of both parameters without requiring separate instruments, multiple sample preparations, or sequential experiments, thereby reducing overall complexity while maintaining measurement precision.
Solution Approach 2:
The system enables continuous monitoring of both binding events and subsequent enzyme reactions in real-time without interrupting the sample flow or requiring intermediate processing steps. The continuous action maintains measurement precision while simplifying the overall assay process by eliminating discrete measurement steps.
3Ease of operation
If conventional SPR detection is used, then the method is label-free and simple, but it cannot assess the functional activity or enzyme reaction products
Solution Approach 1:
The sensor chip is segmented into specialized regions: the first reaction region maintains label-free binding detection capability, while the second reaction region is specifically designed for detecting enzyme reaction products. This segmentation preserves the simplicity of label-free SPR detection while adding functional activity assessment capability.
Solution Approach 2:
The biosensor system achieves multi-functionality by integrating both binding detection and enzyme activity detection in a single platform. It maintains the advantage of label-free operation for binding detection while simultaneously providing functional activity information through product detection in the second reaction region.
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
Enables the detection of compound binding to functional proteins and assessment of activity impacts without labels or fluorescence, enhancing the sensitivity and specificity of protein interaction analysis.
Implementation Method 1
a method for detecting a specific binding based on changed refractive indexes via surface plasmon resonance (SPR)
Implementation Method 2
a light source means for irradiating the back surface of the thin metal layer via total internal reflection through the prism
Data Source
AI summary
An object of the present invention is to provide a biosensor that can detect binding of a compound with a functional protein and then assay a reaction product derived from the activity of the functional protein again. The present invention provides a biosensor for detecting a test molecule specifically binding to a physiologically active substance, which comprises; (1) (a) a first reaction region on which the physiologically active substance has been immobilized for performing a binding reaction between the physiologically active substance and the test molecule and a physiologically active reaction caused by the physiologically active substance, and (b) a second reaction region on which a molecule that specifically binds to a reaction product resulting from the physiologically active reaction has been immobilized for performing a binding reaction between the reaction product and the molecule that specifically binds to the reaction product, in the same area; and (2) an assay region for detecting changes in the binding reaction in the first reaction region and in the binding reaction in the second reaction region.