Multi-chromophoric Quencher Constructs for Broad Absorption
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Solution Overview
Problem
Current fluorescence quenchers have low absorptivity and a limited range of absorption, which restricts their quenching capacity and effectiveness in fluorescence detection assays, necessitating the development of quenchers with higher absorptivity and broader absorption ranges.
Innovation Solution
The creation of multi-chromophoric quenchers, which comprise multiple dark quenching moieties linked by a multivalent linker, enhances absorptivity and expands the absorption range, allowing for more efficient energy transfer and broader quenching capabilities.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If multiple dark quenching moieties are linked together by a multivalent linker to form multi-chromophoric quenchers, then absorptivity and absorption range are enhanced, but device complexity increases
Solution Approach 1:
Multiple dark quenching moieties are merged into a single multi-chromophoric quencher molecule through covalent linkage by multivalent linkers. This combining approach increases the total absorptivity and broadens the absorption range by integrating multiple chromophores that absorb at different wavelengths, directly resolving the contradiction between enhancing absorptivity and maintaining structural simplicity.
Solution Approach 2:
The patent creates composite quencher structures by combining multiple different dark quenching moieties with distinct absorption characteristics into a single hybrid molecule. This composite approach allows the quencher to exhibit enhanced and broadened absorption properties that exceed those of individual monomeric quenchers, effectively resolving the technical contradiction.
2Adaptability or versatility
If multiple different quenching moieties are combined to expand absorption range, then quenching capability across broader wavelengths is improved, but manufacturing complexity increases
Solution Approach 1:
The multi-chromophoric quencher is synthesized through segmented assembly of individual dark quenching moieties connected by multivalent linkers. This modular segmentation allows each quenching moiety to be independently selected and positioned to target specific wavelength ranges, facilitating systematic design and manufacturing while achieving broad absorption coverage.
Solution Approach 2:
The multivalent linker serves multiple functions: it covalently connects multiple quenching moieties, positions them at optimal distances for energy transfer, and provides structural stability. This multi-functionality simplifies the overall manufacturing process by using a single linker component to achieve several structural and functional requirements simultaneously.
3Quantity of substance
If more quenching moieties are added to increase absorptivity, then quenching capacity is enhanced, but the point of diminishing return is reached
Solution Approach 1:
The patent applies partial action by incorporating a specific number of quenching moieties (typically 2-4) into each multi-chromophoric quencher, rather than adding excessive numbers. This optimized quantity achieves sufficient absorptivity enhancement and quenching capacity while avoiding the diminishing returns and increased complexity associated with adding too many moieties, thus maintaining assay efficiency.
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 increases the sensitivity and dynamic range of assays by enhancing the quenching capacity and absorption range of fluorescence quenchers, enabling them to effectively quench a broader range of reporter emissions with improved sensitivity.
Implementation Method 1
A change in fluorescence, as a result of a change in quenching, occurs upon interaction with a target. Unfortunately, quenchers available to date tend to have low absorptivity as measured by their extinction coefficients. Often, the extinction coefficients are less than 50,000 and more typically less than 30,000, although some quenchers with higher extinction coefficients are available. A quencher with low absorbitivity will have a low quenching capacity of a reporter by Förster energy transfer.
Data Source
AI summary
Dark quencher constructs, termed “multi-chromophoric quenchers” are described herein that comprise at least two dark quenching moieties, which can be the same or different, linked together by at least one multivalent linking moiety. The structure of the multi-chromophoric quenchers can be varied to selectively enhance quenching within a specific range of reporter emission wavelengths. This can be accomplished by linking together, into a single molecule, two or more identical quenchers, by reacting the quenchers with a multivalent linker. The structure of the multi-chromophoric quencher can also be varied to quench a broader range of reporter emission wavelengths than previously possible. This can be accomplished by linking together, into a single molecule, two or more different quenchers, by reacting the quenchers with a multivalent linker. The structure of the multi-chromophoric quencher can also be varied to simultaneously broaden the absorption range and increase the total absorption within the absorption range. This can be done by combining the two concepts described above. In other words, multiple types of quenching moieties can be employed to increase the absorption range and a multiple number of each type of quenching moiety can be used to increase the total absorptivity within the absorption range. The multi-chromophoric quenchers can be tethered to probes for biomolecules, insoluble supports and/or fluorescent dyes for use in a wide variety of biomolecular assays.