Dye-Conjugated Reagent Drying for Multiplex Assay Stability
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Solution Overview
Problem
Existing multiplexed assays face issues with cross-linking of labeled reagents during the drying process, leading to erroneous results, especially in flow cytometry, due to the chemical nature of polymeric dyes which bind with each other, causing anomalous cell populations and variability in multi-parameter studies, and resulting in unstable reagents that cannot be stored for long periods.
Innovation Solution
The method involves drying down dye-conjugated reagents in separate locations on substrates to prevent interaction, allowing them to remain stable and maintain specific binding to target analytes, with the reaction vessel configured to receive a liquid sample that dissolves the binding partners, ensuring accurate multiplex binding assays without cross-linking.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If dye-conjugated reagents are dried down in aqueous solution for stable storage, then the reagents become stable at room temperature and can be stored for extended periods, but the dyes become cross-linked and bind to each other, causing erroneous results
Solution Approach 1:
The patent segments the drying process by using separate substrate locations for different dye-conjugated reagents. Each reagent is dried at a distinct location on the substrate, preventing physical contact and cross-linking between different dyes during the drying and storage process. This spatial segmentation allows stable storage while maintaining assay accuracy.
Solution Approach 2:
The substrate acts as an intermediary medium that enables separate storage of multiple dye-conjugated reagents. By depositing each reagent at a distinct substrate location, the substrate provides a stable platform that prevents direct interaction between dyes while allowing all reagents to be stored together in a single container.
2Ease of operation
If multiple polymeric dyes are formulated in the same solution for multiplex assays, then convenience is improved, but the dyes bind to each other through hydrophobic bonds, causing erroneous compensation settings and anomalous cell populations
Solution Approach 1:
The patent applies segmentation by providing separate substrate locations for each dye-conjugated reagent. This spatial separation prevents hydrophobic bonding between different polymeric dyes while allowing all reagents to be stored in a single container, maintaining both convenience and measurement precision.
Solution Approach 2:
The patent implements local quality by creating distinct micro-environments at different substrate locations. Each location contains a specific dye-conjugated reagent in a controlled local environment, preventing unwanted interactions between different dyes while maintaining the overall convenience of a single-container system.
3Reliability
If dye-conjugated reagents are stored separately and formulated prior to experiments, then cross-linking is prevented, but additional variability is introduced in multi-parameter studies and reagent wastage occurs
Solution Approach 1:
The patent merges the benefits of separate storage with the convenience of single-container formulation. Multiple dye-conjugated reagents are deposited at separate substrate locations and stored together in one container, eliminating the need for multiple separate storage containers while preventing cross-linking through spatial separation.
Solution Approach 2:
The patent uses segmentation at the substrate level to maintain reagent stability while enabling combined storage. Each reagent remains physically separated at its designated substrate location, preventing cross-linking, while all reagents can be accessed together in a single container, reducing protocol complexity.
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 stabilizes labeled reagents at room temperature for extended periods, reducing erroneous results and variability, and allows for long-term storage without refrigeration, improving the accuracy and reliability of multiplex assays.
Implementation Method 1
Such contact results in dissolution of the first and second binding partners to form a liquid reaction mixture comprising the first binding partner and the second binding partner in the reaction vessel
Data Source
AI summary
The method involves drying down dye-conjugated reagents in separate locations in a reaction vessel so that the dyes don't non-specifically interact with each other during drying. This invention thus improves multiplex binding assays by eliminating erroneous results caused by dyes' being non-specifically attached to each other when dried down together.