Autofluorescence-Based Immune Response Evaluation Method
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Solution Overview
Problem
Current methods for evaluating antigen-specific immune responses, such as ELISpot and FluoroSpot assays, are cumbersome, time-consuming, and require cell cultivation, making them unsuitable for quick and efficient testing, especially in infectious disease diagnosis and drug development.
Innovation Solution
A method utilizing autofluorescence changes in cells after antigen stimulation, measured by comparing autofluorescence information between a sample with a test substance and a control sample, allowing for rapid evaluation of immune responses based on the FAD/NADH ratio, which correlates with antigen-specific immune responses.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If ELISpot assay or FluoroSpot assay is used to evaluate antigen-specific immune response, then measurement precision is improved, but loss of time increases and device complexity increases
Solution Approach 1:
The patent extracts the evaluation of immune response from the complex multi-step ELISpot/FluoroSpot workflow by focusing specifically on autofluorescence changes as a standalone measurable parameter. This extraction allows direct measurement of immune cell activation without requiring antigen presentation, cytokine secretion, or complex visualization steps, thereby reducing testing time while maintaining evaluation accuracy
Solution Approach 2:
The patent replaces the mechanical/cellular processes of antigen presentation, immune cell activation, and cytokine secretion with a direct optical measurement of autofluorescence. By substituting the biological cascade with an optical detection method, the system achieves rapid immune response evaluation without the time-consuming cellular interactions required by conventional assays
2Measurement precision
If ELISpot assay or FluoroSpot assay is used to evaluate antigen-specific immune response, then measurement precision is improved, but device complexity increases
Solution Approach 1:
The patent extracts the essential measurement signal (autofluorescence) from the complex ELISpot/FluoroSpot assay workflow, eliminating the need for antigen preparation, cell culture, cytokine capture, and fluorescent labeling steps. This extraction simplifies the assay to a direct optical measurement while preserving the ability to detect immune response
Solution Approach 2:
The patent utilizes the cell's own autofluorescence as the measurement signal, eliminating the need for external fluorescent labels, antibodies, or complex reagents. The immune cells themselves provide the measurement signal through their intrinsic autofluorescence properties, thereby simplifying the assay procedure and reducing operational complexity
3Measurement precision
If cell cultivation is performed until immune effector molecules are secreted, then measurement precision is improved, but loss of time increases
Solution Approach 1:
The patent performs preliminary measurement of autofluorescence before immune effector molecules are secreted during cell cultivation. By measuring the baseline autofluorescence signal in advance and comparing it with post-stimulation signals, the system can detect immune response activation without requiring the time-consuming secretion and accumulation of cytokines
Solution Approach 2:
The patent substitutes the time-dependent cellular process of cytokine secretion with an immediate optical measurement of autofluorescence changes. This substitution replaces the slow biological cascade with rapid photon detection, achieving both high sensitivity and speed without requiring extended cultivation periods
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 a simple, quick, and efficient assessment of immune responses, reducing testing time to less than 1 hour and avoiding the need for extensive cell cultivation, while maintaining high sensitivity and specificity.
Implementation Method 1
autofluorescence of the cell, namely, fluorescence emitted from an endogenous fluorescent substance possessed by the cell when the cell is irradiated with excitation light having a specific wavelength, changes in a short time after the stimulation with the antigen
Data Source
AI summary
A method of evaluating an immune response of a cell group to a test substance, the method comprising evaluating the immune response of the cell group to the test substance based on a comparison between autofluorescence information of a first sample prepared by adding the test substance to the cell group and autofluorescence information of a second sample, the second sample serving as a control for the first sample.