Homogeneous CRET Immunosensor for Rapid Low-Concentration Detection
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Solution Overview
Problem
Current methods for detecting analytes in samples, such as enzyme-linked immunosorbent assays, are laborious, prone to operator error, and require multiple wash cycles, and biosensors lack sensitivity and selectivity for detecting analytes at low concentrations.
Innovation Solution
The use of chemiluminescence resonance energy transfer (CRET) pairs, comprising an antibody or antibody-like molecule attached to a first component and a labelled antigen to a second component, where energy transfer efficiency between 10 to 75% indicates the presence of an analyte, allowing for real-time detection without secondary antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If heterogeneous assay formats with immobilization on solid surface are used, then detection can be performed, but the process becomes time laborious and prone to operator error due to multiple wash cycles and incubation steps
Solution Approach 1:
The invention extracts the detection system from the conventional heterogeneous solid-surface format and transforms it into a homogeneous solution-based format. The CRET pair components remain in solution throughout the assay, eliminating the need for immobilization, washing, and multiple incubation steps while maintaining detection capability.
Solution Approach 2:
The invention introduces a CRET pair as an intermediary detection system. The first component (donor) and second component (acceptor) serve as mediators that transfer energy when in proximity, enabling detection through energy transfer efficiency measurements rather than traditional signal amplification methods.
2Speed
If conventional biosensors are used for analyte detection, then rapid response can be achieved, but sensitivity and selectivity for detecting analytes at low concentrations remain insufficient
Solution Approach 1:
The invention changes the detection parameter from traditional signal intensity measurement to energy transfer efficiency measurement. By monitoring the efficiency of energy transfer between CRET pair components, the system achieves enhanced sensitivity for detecting analytes at low concentrations while maintaining rapid response characteristics.
3Measurement precision
If multiple wash cycles and incubation steps are performed, then detection accuracy can be maintained, but operator error increases and automation becomes more difficult
Solution Approach 1:
The invention removes the multiple wash cycles and separate incubation steps from the assay protocol. The detection is performed in a single homogeneous solution phase, eliminating the mechanical operations that are prone to operator error and difficult to automate.
Solution Approach 2:
The invention merges multiple separate operations (immobilization, washing, incubation) into a single homogeneous solution-based detection step. All components interact in solution simultaneously, simplifying the protocol and reducing opportunities for operator error.
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 CRET-based method provides increased sensitivity and shorter assay times, enabling rapid and specific detection of a wide range of analytes without the need for multiple wash cycles or immobilization on a solid surface.
Implementation Method 1
contacting the sample with: an antibody or antibody-like molecule capable of binding to the analyte attached to a first component of a chemiluminescence resonance energy transfer (CRET) pair; and a labelled antigen comprising an antigen capable of binding to the antibody or the antibody-like molecule attached to a second component of the CRET pair
Data Source
AI summary
The present invention relates to methods for detecting analytes in a sample. The methods may also be used to determine the amount of analyte in the sample. The present invention also relates to antibody or antibody-like molecules and labelled antigens for use in these methods.