Solid Phase Assay for Multiple Analyte Ratio Quantification
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Solution Overview
Problem
Current immunoassays face limitations in accurately determining the relative amounts of multiple analytes in fluid samples due to constraints on particle movement and background signals, as well as interference from related analytes, making it difficult to assess the presence of multiple analytes of interest simultaneously.
Innovation Solution
A method using a solid phase assay with multiple sample capture zones and analyte binding particles coated with specific binding agents, allowing the analyte binding particles to bind to their respective capture reagents, enabling the determination of the ratio of analyte binding particles to quantify the relative amounts of analytes without the need for internal controls.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If traditional immunoassays are used to detect multiple analytes, then the assay can identify the presence of analytes, but particle movement constraints and background signals reduce measurement precision and reliability
Solution Approach 1:
The assay system is segmented into distinct functional components: analyte-specific binding particles, multiple sample capture zones with specific capture reagents, and separate detection zones. This segmentation allows independent optimization of each component, reducing cross-interference and improving measurement precision for multiple analytes simultaneously
Solution Approach 2:
Analyte-specific binding particles serve as intermediaries that bridge the analyte in the sample and the solid phase capture reagents. These particles facilitate specific analyte capture while minimizing background signals, thereby improving both measurement precision and assay reliability
2Adaptability or versatility
If related analytes are present in the sample, then the assay detects their presence, but they compete or interfere with each other, making it difficult to assess the correct presence of multiple analytes
Solution Approach 1:
Different sample capture zones are designed with specific local qualities - each zone contains capture reagents specific to a particular analyte. This local specialization ensures that related analytes are captured by their respective specific reagents rather than competing for the same binding sites, maintaining measurement precision while enabling multi-analyte detection
Solution Approach 2:
The detection system is segmented into analyte-specific pathways, where each analyte follows a distinct route from sample application through its specific capture zone to detection. This segmentation eliminates cross-interference between related analytes while preserving the system's ability to detect multiple analytes simultaneously
3Reliability
If internal controls are used in quantitative determination, then the assay can account for variability, but the device complexity and procedure steps increase
Solution Approach 1:
The assay system performs self-calibration and self-validation through the competitive binding mechanism itself. The binding particles and capture reagents automatically establish equilibrium relationships that provide intrinsic quantitative information, eliminating the need for separate internal controls while maintaining reliability
Solution Approach 2:
The system uses changes in binding parameters (binding affinity, concentration ratios) as inherent references for quantification. By measuring the competitive binding equilibrium, the system derives quantitative information without requiring additional control reagents or procedural steps, thus reducing complexity while maintaining accuracy
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 method provides a simple and highly accurate assessment of relative analyte amounts, eliminating the need for internal controls and reducing errors associated with standard curve interpolation, while effectively distinguishing between multiple analytes of interest.
Implementation Method 1
Immunoassays utilize the specificity of the antigen (Ag) - antibody (Ab) reaction to detect an Ag or Ab in a sample
Implementation Method 2
one reagent (e.g., the Ag or Ab) is attached to a solid surface, facilitating separation of bound reagents or analytes from free reagents or analytes
Data Source
AI summary
Methods for measuring the relative amount of two or more analytes of interest in a fluid sample, as well as kits useful in the methods, are disclosed. The methods involve assays that utilize a solid phase apparatus with a membrane having an application point and at least two sample capture zones having sample capture reagents; analyte binding particles or analyte coated particles; and assessment of a ratio of such particles arrested in capture zones, wherein the ratio is equal to, or inversely equal to, the relative amounts of the analytes of interest in the fluid sample.