FRET Signal Suppressor Agents for Homogeneous Multiplexing
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Solution Overview
Problem
Current multiplexing techniques for biological assays require complex equipment and often involve separation or washing steps, making them unsuitable for high-throughput screening in the pharmaceutical industry, particularly for detecting biomolecules inside intact cells or in a homogeneous medium.
Innovation Solution
A method that suppresses the FRET signal by introducing FRET signal suppressor agents, which do not disturb the biological event, allowing for multiplexing in a homogeneous medium without the need for separation or washing steps and using standard fluorimeters, enabling the detection of multiple biological events with existing equipment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If current multiplexing techniques are used, then multiple biological events can be detected, but complex equipment and separation/washing steps are required
Solution Approach 1:
The patent introduces FRET signal suppressor agents as intermediary substances that selectively interfere with FRET signals between specific fluorophore pairs. These suppressors act as mediators that allow discrimination between multiple biological events without requiring complex equipment, by chemically interfering with the energy transfer process rather than relying on sophisticated optical discrimination systems
Solution Approach 2:
The patent changes the detection parameter from optical wavelength discrimination (requiring complex instruments) to FRET signal intensity measurement (usable with standard fluorimeters). By monitoring changes in FRET signal strength rather than emission wavelengths, the system achieves multiplexing with simpler equipment
2Adaptability or versatility
If current multiplexing techniques are used, then multiple biological events can be detected, but separation or washing steps are required
Solution Approach 1:
The patent extracts the FRET signal suppression function from complex separation processes. Instead of physically separating multiple biological events through washing steps, the system chemically extracts and suppresses specific FRET signals using suppressor agents, allowing all events to remain in the same homogeneous medium while enabling selective detection
Solution Approach 2:
FRET signal suppressor agents serve as intermediary substances that enable selective signal suppression without requiring physical separation. These chemical mediators interact specifically with fluorophore pairs to suppress their FRET signals, eliminating the need for mechanical separation steps
3Adaptability or versatility
If FRET signal suppressor agents are introduced, then FRET signal can be suppressed for multiplexing, but the agents must not disturb the biological event
Solution Approach 1:
The patent applies local quality by ensuring that FRET signal suppressors interact selectively with specific fluorophore pairs involved in FRET, while leaving other biological components unaffected. The suppressors exhibit specific affinity for certain fluorophores, creating localized chemical interactions that suppress FRET signals without disturbing the overall biological event being studied
Solution Approach 2:
FRET signal suppressors act as intermediary substances that intervene in the FRET process without interfering with the biological event. These agents mediate between the fluorophores and the detection system, selectively suppressing FRET signals while maintaining the integrity of the biological molecules and processes under investigation
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 efficient multiplexing of biological events in a homogeneous medium without complex equipment or separation steps, allowing for the detection of multiple biomolecules or events simultaneously, enhancing the capability for high-throughput screening and pharmaceutical applications.
Implementation Method 1
The invention relates to the suppression of a FRET signal, or fluorescence resulting from a transfer of resonance energy
Data Source
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AI summary
The invention relates to a method for multiplexing biological events in a biological sample, characterized in that it comprises the suppression of the FRET emitted by the reaction medium containing at least two pairs of fluorescent FRET partner conjugates, each being specific for one of the biological events studied. This suppression of the emitted FRET is carried out by introducing into the medium a FRET signal suppressor agent characterized in that it has a domain for binding to a fluorophore which is a member of a pair of FRET partners and is capable of suppressing the FRET signal emitted when the two partners are in proximity to one another.