Dual Capture Reagent Assay for Protein Concentration
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Solution Overview
Problem
Current methods for determining protein concentrations in biological samples with proteins expressed at widely diverging levels (at least 10-fold) are inadequate, as they require sample dilution, which lowers the concentration of low-concentration analytes below the limit of detection.
Innovation Solution
The method involves using an assay plate with first and second capture reagents bound to surfaces, where the first capture reagents selectively bind to low-concentration proteins and the second capture reagents bind to high-concentration proteins, allowing for the determination of both protein concentrations in the same sample well.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If sample dilution is performed to ensure highly secreted proteins are in the linear range of detection, then the measurement precision of high concentration proteins is improved, but the concentration of low concentration analytes falls below the lower limit of detection
Solution Approach 1:
The assay is divided into two separate capture stages: first capture reagents bind low concentration proteins, then second capture reagents bind high concentration proteins. This segmentation allows each capture reagent to be optimized for its specific target concentration range, resolving the contradiction between detecting high and low concentration proteins simultaneously
Solution Approach 2:
Different capture reagents with different binding affinities and specificities are applied to different protein targets within the same sample. The first capture reagents are designed with high affinity for low concentration proteins, while second capture reagents are designed for high concentration proteins, creating local optimization for each detection target
2Loss of substance
If a single assay is used to measure both low and high concentration proteins, then reagent consumption is reduced, but the dynamic range of detection becomes insufficient
Solution Approach 1:
The assay system performs multiple functions within a single workflow: it detects both low and high concentration proteins, maintains appropriate dynamic range for each, and reduces overall reagent consumption by processing all targets in one sample preparation sequence
Solution Approach 2:
The assay changes detection parameters by using different capture reagents with different binding characteristics for different protein concentration ranges. This allows the system to adapt its detection parameters to match the specific concentration range of each target protein while maintaining a unified assay protocol
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 enables accurate determination of both low and high concentration proteins in the same sample, improving the dynamic range and reducing reagent consumption, thereby overcoming the limitations of existing methods.
Implementation Method 1
the first binding molecules selectively bind to the at least one low concentration protein
Implementation Method 2
the second binding molecules selectively bind to the at least one high concentration protein
Data Source
AI summary
Disclosed herein are methods for determining a concentration of at least one low concentration protein and at least one high concentration protein in a biological sample.