Microfluidic Assay Cartridge Segmentation for Multiplexing

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Solution Overview

Problem

Multiplexed assays face challenges due to biological cross-reactivity and sub-optimal buffer conditions, leading to compromised data quality and impractical design requirements as the number of variables increases.

Innovation Solution

A microfluidic assay device with separate, fluidically isolated channels and functionalized hollow elements allows for precise control of sample and reagent flow, using capture moieties and optimized buffer conditions to minimize cross-reactivity and automate manual steps, enabling improved data quality and ease of use.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If multiple analytes and detection cocktail are mixed in a single reaction vessel to perform multiplexed assays, then the multiplex level increases, but biological cross-reactivity occurs causing unintended secondary reactions that distort measurements and compromise data quality

Engineering Contradiction:
Improvemultiplex levelVSAvoiddata quality
Core Design Contradiction:
Adaptability or versatilityVSMeasurement precision

Solution Approach 1:

The invention divides the single reaction vessel into multiple separate reaction vessels, each containing a single analyte and its specific detection reagents. This segmentation prevents biological cross-reactivity between different analyte-detection reagent combinations while maintaining the ability to perform multiple assays simultaneously, thus preserving data quality while achieving multiplexing capability.

Inventive Principle:
Principle #1Segmentation

2Device complexity

If a common assay buffer is applied across all antibodies in the multiplexed system, then the assay design becomes simpler, but the performance of individual components deteriorates because the common buffer is not optimal for each specific antibody

Engineering Contradiction:
Improveassay design complexityVSAvoidcomponent performance
Core Design Contradiction:
Device complexityVSMeasurement precision

Solution Approach 1:

The invention assigns different optimized buffers to each separate reaction vessel based on the specific requirements of each antibody-analyte combination. This local optimization allows each assay to perform at its best with buffer conditions tailored to its specific components, while the overall system remains manageable through modular design where each vessel is independently configured.

Inventive Principle:
Principle #3Local quality

3Adaptability or versatility

If the number of variables in the multiplexed system increases to accommodate more analytes, then the multiplex level increases, but the assay design becomes increasingly impractical and difficult

Engineering Contradiction:
Improvemultiplex levelVSAvoidassay design complexity
Core Design Contradiction:
Adaptability or versatilityVSDevice complexity

Solution Approach 1:

By segmenting the assay into separate reaction vessels, each handling a single analyte with its own optimized buffer and detection reagents, the invention reduces the complexity of designing and managing multiple variables. Each vessel becomes an independent, simplified unit that can be designed and optimized separately, making the overall multiplexed system more practical and manageable.

Inventive Principle:
Principle #1Segmentation

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 solution provides low-cost, fast reaction times, high sensitivity, and large dynamic range, significantly improving data quality and ease of use in multiplex assays while minimizing the need to design around cross-reactivity.

Implementation Method 1

The at least one hollow element may be functionalized with a capture moiety or molecules so as to form at least one reaction vessel

Methodology Applied
Scientific EffectMolecular recognition:

Implementation Method 2

The microfluidic channels and micro-valves may be configured to respond to signaling containing information about performing the assay and to controllably receive the sample and at least one reagent in the at least one reaction vessel

Methodology Applied
Scientific EffectPressure differential: Pressure Gradient

Implementation Method 3

reagents, including an enzymatic substrate, for producing an emitted light signal

Methodology Applied
Scientific EffectEnzymatic reaction: Enzyme

Implementation Method 4

producing an emitted light signal

Methodology Applied
Scientific EffectLight emission: Luminescence

Implementation Method 5

introduce a wash solution to remove any non-specifically bound proteins or antibodies

Methodology Applied
Scientific EffectDifferential solubility:

Data Source

PatentEP2504701B1Method and apparatus for performing assays
Publication Date: 2017.09.13 CYVEK INC
  • EP2504701B1 patent drawingFigure 1~1(b)
  • EP2504701B1 patent drawingFigure 1(c)
  • EP2504701B1 patent drawingFigure 2

AI summary

An apparatus is provided for performing an chemical, biochemical, or biological assay on a sample comprising: a microfluidic assay cartridge (1) that contains at least one sample inlet well (2) configured to receive a sample; and a microfluidic sub-unit (3) associated with the microfluidic assay cartridge (1) and comprising microfluidic channels (8), micro-valves (4, 4a, 9) and at least one separate and fluidicly-isolated isolation channel (5), and at least one hollow element (14); the at least one hollow element (14) being functionalized with a capture moiety or molecules (15) so as to form at least one reaction vessel (19); the microfluidic channels (8) and micro-valves (4, 4a, 9) configured to respond to signaling containing information about performing the assay and to controllably receive the sample and at least one reagent in the at least one reaction vessel (19), and to provide from the at least one reaction vessel (19) light containing information about the assay performed on the sample inside the at least one reaction vessel (19) as a result of said at least one reagent.