Concentric FRET Relay for Parallel Enzyme Detection

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Solution Overview

Problem

Conventional Förster resonance energy transfer (FRET) configurations are limited in extending energy transfer distances and require multiple discrete donor-acceptor pairs, which complicates multiplexed detection in biological applications.

Innovation Solution

A concentric FRET relay assembly featuring a semiconductor quantum dot as the initial donor and two fluorescent dyes, Alexa Fluor 555 and Alexa Fluor 647, assembled at similar distances, allowing for three energy transfer pathways, including a two-step relay and a direct energy transfer pathway, enabling parallel detection of bio/physicochemical processes with a single probe entity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Length of stationary object

If multiple discrete donor-acceptor FRET pairs are used to extend energy transfer distance, then the energy transfer distance is extended, but the device complexity increases

Engineering Contradiction:

Improveenergy transfer distanceVSAvoiddevice complexity

Core Design Contradiction:

Length of stationary objectVSDevice complexity

Solution Approach 1:

The patent implements a nested FRET relay configuration where a first FRET pair (donor1-acceptor1) and a second FRET pair (donor2-acceptor2) are arranged concentrically around a central acceptor. The inner FRET pair is positioned within the structure of the outer FRET pair, creating a nested architecture that extends energy transfer distance while maintaining structural compactness and reducing overall device complexity compared to linear arrangements.

Inventive Principle:

Principle #7Nested doll (Nesting)

2Adaptability or versatility

If multiple discrete FRET pairs are used for multiplexed detection, then the detection capability is improved, but the ease of operation deteriorates

Engineering Contradiction:

Improvedetection capabilityVSAvoidease of operation

Core Design Contradiction:

Adaptability or versatilityVSEase of operation

Solution Approach 1:

The patent combines multiple FRET pairs into a single integrated concentric assembly where all components (donor1, acceptor1, donor2, acceptor2) are positioned in close proximity around a central point. This merging of multiple detection functions into one unified structure simplifies operation by eliminating the need to handle separate probes, while maintaining the versatility to detect multiple bio/physicochemical processes simultaneously through distinct FRET pathways.

Inventive Principle:

Principle #5Merging (Combining)

3Ease of manufacture

If discrete FRET pairs are used, then the manufacturing process is simplified, but the measurement precision for multiple processes deteriorates

Engineering Contradiction:

Improvemanufacturing processVSAvoidmeasurement precision

Core Design Contradiction:

Ease of manufactureVSMeasurement precision

Solution Approach 1:

The patent employs local quality by assigning specific functional roles to different components at different spatial locations within the concentric structure. Each donor-acceptor pair is positioned with specific distance relationships (e.g., acceptor1 at distance d1 from donor1, acceptor2 at distance d2 from donor2) to optimize FRET efficiency for specific detection purposes. This localized optimization of structural parameters enables precise measurement of multiple processes while maintaining a manufacturable design through standardized modular components.

Inventive Principle:

Principle #3Local quality

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 quantitative measurement of two proteolytic enzyme activities in parallel using a single probe entity, providing two analytic optical signals from a single entity, overcoming the limitations of discrete FRET pairs and extending energy transfer distances.

Implementation Method 1

Conventional Förster (or fluorescence) resonance energy transfer (FRET) configurations typically comprise two fluorescent dyes assembled into discrete pairs. Energy transfer is observed in a single step from the donor to the acceptor.

Methodology Applied

Scientific EffectFörster resonance energy transfer (FRET):

Implementation Method 2

More complex FRET configurations with multiple energy transfer steps have also been described, where energy is transferred from an initial donor to a terminal acceptor in n successive steps through n−1 intermediary dyes that act as both an acceptor (for the previous dye in the sequence) and donor (for the next dye in the sequence).

Methodology Applied

Scientific EffectFörster resonance energy transfer (FRET):

Implementation Method 3

a second fluorescent dye configured as a FRET acceptor in both (a) a second FRET process wherein the first fluorescent dye is a donor, and (b) a relatively inefficient third FRET process wherein the QD is a donor

Methodology Applied

Scientific EffectFörster resonance energy transfer (FRET):

Data Source

PatentUS9120967B2Concentric forster resonance energy transfer relay for the parallel detection of two bio/physicochemical process

Publication Date: 2015.09.01 THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY DEPARTMENT OF HEALTH & HUMAN SERVICES

AI summary

Described herein is a Förster (or fluorescence) resonance energy transfer (FRET) configuration with three energy transfer pathways between three luminescent components, where two of the energy transfer steps occur in sequence as a relay, and the first step of the relay is in competition with a third energy transfer process (energy transfer from the donor to the intermediary is in competition with energy transfer from the donor directly to the terminal acceptor).