Dendritic Signal Amplification in SPR Binding Detection
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Solution Overview
Problem
Current SPR assays face limitations in sensitivity and throughput due to unspecific binding, high reagent consumption, and complex detection configurations, particularly in flow cell formats, which hinder efficient amplification of binding events between analytes and binding agents.
Innovation Solution
The method involves combining multivalent molecules in a fluidic system for dendritic amplification by mixing two enhancement molecules simultaneously before the detection area, allowing for increased amplification efficiency and reduced constraints on detection devices, using a fluidics system with conduits that mix fluids at a junction and direct the mixed fluids over a solid support for surface plasmon resonance detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sequential injection of enhancement reagents is used, then binding events can be detected, but sensitivity is limited due to limited binding sites for enhancement molecules
Solution Approach 1:
The patent combines multiple enhancement reagents (polyclonal antibody and anti-rabbit IgG) into a single simultaneous injection mixture, allowing both reagents to bind to the analyte-antibody complex at the same time. This merging approach creates a dendritic amplification structure that significantly increases the number of available binding sites and enhances detection sensitivity compared to sequential injection methods.
2Measurement precision
If large complexes are used for enhancement, then signal amplification is achieved, but binding capacity to sensor chips with dextran matrix is lost
Solution Approach 1:
The patent changes the physical parameters of the enhancement system by using simultaneously injected multivalent molecules that form dendritic structures in solution rather than pre-formed large complexes. This parameter change maintains binding capacity to the sensor chip surface while achieving signal amplification through the dendritic arrangement of enhancement molecules.
3Measurement precision
If alternately injecting two enhancement molecules is used, then dendritic amplification is achieved, but assay time increases and material consumption increases
Solution Approach 1:
The patent merges the injection of multiple enhancement reagents into a single simultaneous injection event. The mixture of polyclonal antibody and anti-rabbit IgG is injected together, allowing both reagents to bind concurrently and form dendritic structures in a single step, thereby reducing assay time and reagent consumption while maintaining amplification efficiency.
4Measurement precision
If laminar flow-assisted dendritic amplification is used, then signal amplification is achieved, but high demands are placed on flow conditions and detection device choices are limited
Solution Approach 1:
The patent extracts the dendritic amplification process from the complex laminar flow system and implements it in a simpler flow cell configuration. By simultaneously injecting enhancement reagents that form dendritic structures in the flow cell rather than relying on laminar flow merging, the method reduces demands on flow conditions and expands compatibility with different detection devices.
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 achieves significantly amplified detection signals, enabling the detection of trace amounts of analytes with improved sensitivity and reduced reagent consumption, as demonstrated by a 15-fold increase in signal compared to sequential injection methods.
Implementation Method 1
Laminar flow-assisted dendritic amplification is a signal amplification method for biomolecular binding events in microchannels
Implementation Method 2
mixing a solution of a pair of multivalent molecules in a fluidic system and measuring the signal immediately after the two solutions have been substantially mixed
Implementation Method 3
surface plasmon resonance (SPR) by mixing a solution of a pair of multivalent molecules in a fluidic system and measuring the signal immediately after the two solutions have been substantially mixed
Implementation Method 4
dendritic amplification is combined surface plasmon resonance (SPR) by mixing a solution of a pair of multivalent molecules
Implementation Method 5
a pair of multivalent molecules in a fluidic system
Data Source
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AI summary
The present invention relates to a method for detection of binding or interaction events between a binding agent and its corresponding analyte (such as an antibody and an antigen) in which a signal is detected which is substantially more amplified and thus easier to detect than in prior art systems. The method comprises simultaneous but separate addition of a first enhancement reagent having affinity for said analyte and a second enhancement reagent having affinity for the first enhancement reagent wherein the first enhancement reagent binds to the analyte and the second enhancement reagent binds to the first enhancement reagent, and, wherein the first and second enhancement reagents have more than one binding site so that they are able to bind to each other to thereby amplify a detectable signal from the binding event.